Podcasts about HCO

We've got a shockingly low number of proverbial monsters and a perfectly normal amount of hideous creatures on this week's show! Monsters come in many shapes and sizes, but the important thing in this episode is that they come from games with awesome tunes! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:08:55 Game 1 0:11:59 Game 1 Reveal 0:23:00 Game 2 0:31:57 Game 2 Reveal 0:43:01 Lightning Round! 0:50:31 Game 3 0:56:13 Game 3 Reveal 1:10:22 Game 4 1:16:40 Game 4 Reveal 1:31:54 This Game's Winner Is... 1:44:28 Bonus Music/Outro

God plays a massive role in all our lives on a daily basis, but never more importantly than when a game features a prominent god character or lets you play as god. Let us bow our heads in solemn VGM worship as we take a listen to some of our favorite god games and possibly select a new pope in the process! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:07:12 Game 1 0:11:04 Game 1 Reveal 0:25:45 Game 2 0:32:28 Game 2 Reveal 0:49:30 Game 3 0:56:37 Game 3 Reveal 1:11:25 Game 4 1:23:23 Game 4 Reveal 1:39:28 This Game's Winner Is... 1:50:15 Bonus Music/Outro

We're playing the one of the best kinds of bad game on this week's episode: games based on MOVIES! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:09:25 Game 1 0:15:30 Game 1 Reveal 0:31:37 Game 2 0:36:02 Game 2 Reveal 0:49:51 Game That BOOM! 0:57:25 VGM Threesome! 1:04:05 VGM Threesome Song 2 1:12:42 VGM Threesome Song 3 1:22:23 Game 4 1:28:40 Game 4 Reveal 1:34:41 Bluesky Bonus Request! 1:37:05 This Game's Winner Is... 1:42:51 Bonus Music/Outro

We're playing cartoon games on this episode! The games are bad and the tunes are bad, but the memories...well, they hold up better than the games. Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:07:40 Game 1 0:13:21 Game 1 Reveal 0:23:39 Game 2 0:28:46 Game 2 Reveal 0:33:46 Lightning Round! 0:43:01 Game 3 0:47:04 Game 3 Reveal 0:57:11 Game 4 1:03:04 Game 4 Reveal 1:13:36 Lightning Round!! 1:21:44 Game 5 1:26:55 Game 5 Reveal 1:40:14 This Game's Winner Is... 1:49:50 Bonus Music/Outro

*** WELCOME TO OUR 100th Episode of Homeopathy Hangout Podcast! Please support our sponsors on their social links below *** Today's super special guest is Miranda Castro - a British homeopath who has been practicing classical homeopathy since 1982. She is author of the best-selling "The Complete Homeopathy Handbook" (over 500,000 copies sold), the much-loved Homeopathy for Mother and Baby and A Homeopathic Guide to Stress.  Miranda believes passionately that homeopathy is 'medicine for the people' and her writing reflects that: her numerous articles and her books have made classical homeopathy spectacularly accessible to the lay person (home prescriber?)—giving people the tools to accurately select healing remedies for themselves and their families.  She has the ability to communicate homeopathic topics in an engaging and accessible manner. Her infectious enthusiasm comes through in all her articles, lectures, webinars … and podcasts. She moved to the US in 1994 and currently lives in Florida. She can be contacted at mirandacastro.com A big thank you to the sponsors for our 100th episode Celebration Party in Perth today. Please follow/like them and support them if you can. We are so grateful to them for making today's party super special! - A Woman's touch Photography: instagram.com/awomanstouchphotography/or facebook.com/awomanstouchphotography - Bite into Bliss: facebook.com/biteintothebliss or instagram.com/bitein2bliss/ - Classes Cakes and More: facebook.com/classescakesandmore or instagram.com/classescakesandmore/ - Chaga Health: instagram.com/chagahealth_au or Facebook.com/ChagaHealthAustralia - Clean Minds - no socials, but Anna can be reached on 042 69 72 389 and produces healing stories, cleaning services and placenta prints - Dinner Twist: facebook.com/dinnertwist or instagram.com/dinnertwist - Cakealicious: facebook.com/CakealiciousWA or instagram.com/cakealicious58 - De Light Me Cocktail Cookies: instagram.com/de.light_me - Em Energetic: instagram.com/em.energetic/ or facebook.com/em.energetic - IconFit: facebook.com/iconfitaustralia or instagram.com/iconfitaustralia/ - Just Add Health: facebook.com/JustAddHealthStoneville or instagram.com/justaddhealth - Heather & Co Floral Artisans: instagram.com/handco.perth/ or facebook.com/HCo-104044151916915 - Happy Patch Herbal and Healing: facebook.com/happypatchherbals or instagram.com/happy_patch_herbals/ - Little Miss Organic: instagram.com/little_miss_organic or facebook.com/LittleMissOrganic - Little Miss Enjo: instagram.com/little_miss_enjo or facebook.com/LittleMissENJO - Metta Chiropractic: instagram.com/mettachiropractichealth/ or facebook.com/MettaChiro/ - Macrocos Mish Intuitive Healer: facebook.com/MacrocosMish or instagram.com/macrocosmish - Mother Me Postpartum Meal delivery: instagram.com/_motherme/ or facebook.com/mothermeperth/ - Nourished by Broth: facebook.com/Nourishedbybroth/ - Powder Candle: Instagram.com/powdercandleaustralia or facebook.com/PowderCandleAustralia - The Milk Bar Candle Company:facebook.com/The-Milk-Bar-Candle-Co-102558728992862 or instagram.com/themilkbarcandleco - The Conscious Caregiver: instagram.com/consciousmothersgroup If you would like to support the Homeopathy Hangout Podcast, please consider making a donation by visiting www.EugenieKruger.com and click the DONATE button

We tried a show with no violence, so now it's time for INCREDIBLE VIOLENCE! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:05:06 Game 1 0:16:06 Game 1 Reveal 0:27:57 Game 2 0:33:51 Game 2 Reveal 0:51:10 Game 3 0:55:31 Game 3 Reveal 1:15:43 Game 4 1:23:01 Game 4 Reveal 1:43:44 This Game's Winner Is... 1:50:52 Bonus Music/Outro

Everyone loves to blame violent video games for all the ills that have befallen our society, so in this episode we self-censor and play strictly non-violent games! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:06:25 Game 1 0:11:16 Game 1 Reveal 0:25:30 Game 2 0:30:06 Game 2 Reveal 0:41:04 Game 3 0:45:55 Game 3 Reveal 1:02:58 Game 4 1:08:34 Game 4 Reveal 1:17:23 Game 5 1:22:09 Game 5 Reveal 1:31:52 This Game's Winner Is... 1:38:42 Bonus Music/Outro

We were gonna climb some mountains on this episode but then John went sledding in Tallahassee and got his fill so we're doing a FREEPLAY. Also Johnny went to Magfest, so be prepared to hear about that! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:06:44 Game 1 0:12:05 Game 1 Reveal 0:24:08 Game 2 0:40:02 Game 2 Reveal 0:50:40 Game 3 0:56:19 Game 3 Reveal 1:11:16 Game 4 1:17:14 Game 4 Reveal 1:39:46 Game 5 1:47:57 Game 5 Reveal 2:04:23 This Game's Winner Is... 2:12:30 Bonus Music/Outro

It's a collect-a-thon on this weeks episode! If it's a game with things to collect or even a game collection, it could be on this show! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:05:06 Game 1 0:11:31 Game 1 Reveal 0:25:29 Game 2 0:37:40 Game 2 Reveal 0:57:08 Game 3 1:06:22 Game 3 Reveal 1:21:09 VGM Threesome! 1:23:17 Threesome Game 1 Reveal 1:27:20 Threesome Game 2 1:29:06 Threesome Game 2 Reveal 1:34:40 Threesome Game 3 1:37:18 Threesome Game 3 Reveal 1:44:09 This Game's Winner Is... 1:50:56 Bonus Music/Outro

We're playing complete games on this episode! That means games with no DLC, just the way god intended! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:03:46 Welcome to Game That Tune! 0:10:45 Game 1 0:13:47 Game 1 Reveal 0:32:18 Game 2 0:44:16 Game 2 Reveal 0:57:48 Game 3 1:06:25 Game 3 Reveal 1:24:58 Game 4 1:29:32 Game 4 Reveal 1:44:04 This Game's Winner Is... 1:50:55 Bonus Music/Outro

I'm not a financial advisor; Superpowers for Good should not be considered investment advice. Seek counsel before making investment decisions.Watch the show on television by downloading the e360tv channel app to your Roku, AppleTV or AmazonFireTV. You can also see it on YouTube.When you purchase an item, launch a campaign or create an investment account after clicking a link here, we may earn a fee. Engage to support our work.Devin: What is your superpower?Sharon: One of my staff members said, Sharon, you're the only person who talked about how much joy it brings you to really elevate people and have a team. I hadn't figured it out before, but I think that is my superpower.Asthma and chronic respiratory diseases impact millions globally, limiting lives in profound ways. But Health Care Originals, led by CEO and co-founder Sharon Samjitsingh, is changing the game with a groundbreaking AI-powered wearable device named "Adam." This discreet device listens to lung sounds, predicting and preventing respiratory crises before they happen.Sharon's personal journey with asthma inspired her mission. She recalls, “I almost died five times, went to the ER, or rushed to my doctor's home in the middle of the night. Even though I took my meds, everything was not OK.”Adam combines advanced AI with human touch, offering not just alerts but actionable guidance. “With our Nightingale system, we provide apps, respiratory therapists, and wellness coaches to empower users,” Sharon explains. This holistic approach has yielded remarkable outcomes. Within a year, two-thirds of users report zero attacks, and nearly all experience better symptom management and sleep quality.Health Care Originals is more than a tech innovator; it's a force for change. The company is scaling its Nightingale service, backed by 14 issued patents and a waitlist of 12,000 eager participants. With contracts poised to generate $5.5 million in annual recurring revenue, the firm is expanding its reach across the U.S.Currently raising capital on Wefunder, Sharon invites you to join the journey: “We want everybody to join us on our journey. Be part of helping people live well and breathe easier.”By leveraging innovation and compassion, Sharon is empowering individuals and families to reclaim their lives. This episode underscores how the fusion of technology and empathy can drive meaningful impact.tl;dr:* Sharon Samjitsingh's personal asthma journey inspired her mission to empower respiratory patients.* Health Care Originals developed Adam, an AI-powered wearable to predict respiratory crises.* The Nightingale system includes apps, therapists, and coaches, yielding impressive outcomes.* Sharon's superpower is elevating others by recognizing potential and providing opportunities.* Health Care Originals is raising funds on Wefunder to expand its life-changing solutions.How to Develop Elevating People As a SuperpowerSharon's superpower is her ability to elevate others—whether employees, patients, or collaborators. She combines technical expertise with a profound commitment to helping people succeed, both personally and professionally.Sharon once led a team of 100 shop-floor workers, many with limited education, across 14 manufacturing sites. By training them in root cause analysis and empowering them to present solutions, Sharon helped them save over $1 million in a year. Her efforts boosted their confidence and career trajectories, proving her passion for elevating others.Sharon's tips to develop this superpower:* Recognize Potential: See the strengths in people often overlooked by others.* Empower Through Training: Offer education and opportunities tailored to individual needs.* Support Self-Advocacy: Encourage people to voice their ideas and take ownership of solutions.* Celebrate Success: Acknowledge achievements to build confidence and motivation.By following Sharon's example and advice, you can make elevating people a skill. With practice and effort, you could make it a superpower that enables you to do more good in the world.Remember, however, that research into success suggests that building on your own superpowers is more important than creating new ones or overcoming weaknesses. You do you!Guest ProfileSharon Samjitsingh (she/her):CEO & Co-Founder, Health Care OriginalsAbout Health Care Originals: Health Care Originals (HCO) is a venture-backed healthtech company on a mission to revolutionize the $83B respiratory care market. HCO's AI-powered virtual respiratory care platform and wearable are pioneering always-on, personalized care for the 690M+ sufferers of asthma and chronic obstructive pulmonary disease - the third leading cause of death globally.Backed by leading venture funds Safar Partners and Chloe Capital, and over 1,000 satisfied patients with 12K more on the waitlist, HCO's patented non-invasive solution is ready to improve the quality of life for millions worldwide.Company Facebook Page: fb.com/HealthCareOriginals/Other URL: wefunder.com/hcoBiographical Information:Sharon Samjitsingh, CEO of Health Care Originals (HCO), is a global leader in commercializing innovative technologies across healthcare, sustainability, and manufacturing. With 25+ years of experience, Sharon has managed $150M in innovation projects, holds patents in sustainability and healthcare, and is among fewer than 3,000 professionals with PMP®, PMI-SP®, and PMI-RMP® certifications. Recognized as TechRochester's Technology Woman of the Year (2020) and a Top 100 in Fem Tech & Health Tech honoree (2019), Sharon is a WBL Fellow and an accomplished speaker. She holds an MS from the University of Rochester and a B.Sc. from the University of The West Indies.Linkedin: linkedin.com/in/sharonsamjitsingh/Support Our SponsorsOur generous sponsors make our work possible, serving impact investors, social entrepreneurs, community builders and diverse founders. Today's advertisers include FundingHope, Abby, Motivated Money by Dakin Capital and Honeycomb Credit. Learn more about advertising with us here.Max-Impact MembersThe following Max-Impact Members provide valuable financial support:Carol Fineagan, Independent Consultant | Lory Moore, Lory Moore Law | Marcia Brinton, High Desert Gear | Paul Lovejoy, Stakeholder Enterprise | Pearl Wright, Global Changemaker | Ralf Mandt, Next Pitch | Scott Thorpe, Philanthropist | Add Your Name HereUpcoming SuperCrowd Event CalendarIf a location is not noted, the events below are virtual.* SuperCrowdHour, January 15, 2025, at 1:00 PM Eastern. Devin Thorpe will be doing investor-focused training on building an investment strategy focused on investing in debt instruments to match or even exceed stock market returns. This is great for people serious about investing, whether you are starting with $100 or $100,000. Don't miss it!* Impact Cherub Club Meeting hosted by The Super Crowd, Inc., a public benefit corporation, on January 21, 2024, at 1:00 PM Eastern. Each month, the Club meets to review new offerings for investment consideration and to conduct due diligence on previously screened deals. To join the Impact Cherub Club, become an Impact Member of the SuperCrowd.* SuperCrowdLA: we're going to be live in Santa Monica, California, May 1-3. Plan to join us for a major, in-person event focused on scaling impact. More details to come soon!Community Event Calendar* Successful Funding with Karl Dakin, Tuesdays at 10:00 AM ET - Click on Events* Community Capital Live, Havell Rodrigues, New Majority Capital, January 15, 2:00 PM Eastern.* Motivated Money Method for Raising Funding Workshop, January 16, 11:00 AM - 3:00 PM Eastern.* Community Capital Live, Bernel Hall, New Jersey Community Capital, January 29, 2:00 PM Eastern.* Igniting Community Capital to Build Outdoor Recreation Communities, Crowdfund Better, Thursdays, March 20 & 27, April 3 & 10, 2025, at 1:00 PM ET.* NC3 Changing the Paradigm: Mobilizing Community Investment Funds, March 7, 2025* Asheville Neighborhood Economics, April 1-2, 2-25.If you would like to submit an event for us to share with the 8,000+ members of the SuperCrowd, click here.We use AI to help us write compelling recaps of each episode. Get full access to Superpowers for Good at www.superpowers4good.com/subscribe

Everyone loves paying more after we've already bought a game, so we're discussing games with great DLC and expansions! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:04:45 Game 1 0:11:22 Game 1 Reveal 0:32:25 Game 2 0:44:11 Game 2 Reveal 0:56:13 Game 3 1:17:57 Lightning Round! 1:30:05 Game 4 1:35:34 Game 4 Reveal 1:49:35 Game 5 1:56:07 Game 5 Reveal 2:10:49 This Game's Winner Is... 2:20:26 Bonus Music/Outro

We're playing adventure games on this episode, so we've got epic tunes from epic games! Set your sights on the horizon and enjoy! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:09:41 Game 1 0:19:03 Game 1 Reveal 0:33:37 Game 2 0:40:02 Game 2 Reveal 0:48:33 Game 3 0:51:57 Game 3 Reveal 1:07:15 Game 4 1:21:12 Game 4 Reveal 1:37:00 Game 5 1:45:19 Game 5 Reveal 2:00:13 This Game's Winner Is... 2:04:40 Bonus Music/Outro

We're getting our blood pumping and our balls juiced with HIGH OCTANE GAMES Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters!

Jessie has pulled himself up out of the clitpit and he brought an awesome new mixtape of giant fighting robots with him! Listen to some big time tunes for big time fights! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Bar - MechWarrior 0:00:59 Intro 0:07:45 Dazil, Town of Burning Sands - Xenogears 0:11:05 Manifold Irons - Front Mission (Wonderswan!) 0:13:40 Stage 1-2 (Temple) - Ranger X 0:15:39 The Enemy Commander - Metal Warriors 0:16:38 Gazing Boundary - Ring of Red 0:18:21 Pinnacle Robotics - Into the Breach 0:21:11 The Legendary Mashin - Magic Knight Rayearth 0:23:23 Tomorrow to Start - Vanguard Bandits 0:26:21 Buddies - 13 Sentinels Aegis Rim 0:29:34 Galactica - Zone of the Enders Fist of Mars 0:33:31 Welcome to the Marauder Corps - Titanfall 0:35:18 Cerberus Squad - Chromehounds 0:37:19 Bar( General Purpose) - Front Mission 3 0:40:12 Stage Select - Bangai-O 0:41:21 SUPER 8 Theme - Cyberbots 0:43:21 ZAKU-II - Gundam The Battle Master 2 0:46:22 Pink Ball Activate! - Kirby: Planet Robobot 0:48:36 She's Lost Control - Cyber Troopers Virtual On 0:51:28 Theme from Armored Core 2 - Armored Core 2 0:55:20 Good Feather -Fly a Flag- - Metal Wolf Chaos XD

We've got 5 fantastic games from the year 2003 on this episode! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:04:16 Game 1 0:09:05 Game 1 Reveal 0:23:35 Game 2 0:33:10 Game 2 Reveal 0:47:51 Game 3 0:54:54 Game 3 Reveal 1:16:57 Game 4 1:23:28 Game 4 Reveal 1:37:05 Game 5 1:46:55 Game 5 Reveal 2:01:22 This Game's Winner Is... 2:09:41 Bonus Music/Outro

What is a comeback game? Is it a game on a system that succeeded when its predecessor failed? Is it a game so bad that it got replaced by a completely different game of the same name? Is it a game featuring interdimensional clowns or sword wielding rock stars? The answer may surprise you, so enjoy this episode all about comeback games! Our merch store is back, now on TeePublic! Check it out and grab something absurd for you or a friend! Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters!

We're back after our most absurd August with an all new episode featuring all the games we're playing right now! Or could there be a twist looming? Join us in Discord! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond, HCO and Spanky McMasters! 0:00:00 Welcome to Game That Tune! 0:05:46 VGM Threesome! 0:34:46 Game 2 0:41:19 Game 2 Reveal 0:55:57 Game 3 1:02:44 Game 3 Reveal 1:14:18 Game 4 1:24:04 Game 4 Reveal 1:37:18 Game 5 1:42:05 Game 5 Reveal 1:52:22 This Game's Winner Is... 1:58:48 Bonus Music/Outro

We've got the biggest game yet, featuring the mega-sized ONE HUNDRED THE HARD WAY, Jessie's epic Game That BOOM, and our most absurd finish ever! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:09:09 Game 1 0:12:42 Game 1 Reveal 0:13:48 One Hundred the Hard Way!? 1:29:14 Halftime Show! Game That BOOM returns! 1:35:13 One Hundred the Hard Way Second Half! 1:52:24 Here Comes a New Challenger! 2:34:16 This Game's Winner Is... 2:41:17 Bonus Music/Outro

AUGUST ABSURDITY continues with an epic matchup you'll have to hear to believe! Which John(ny) will come out on top and return to the finals!? Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:05:50 Game 1 0:08:53 Game 1 Reveal 0:22:33 Game 2 0:28:03 Game 2 Reveal 0:40:53 Game 3 0:46:15 Game 3 Reveal 0:56:13 Game 4 1:00:54 Game 4 Reveal 1:08:58 Game 5 1:16:34 Game 5 Reveal 1:27:26 Game 6! 1:33:36 Game 6 Reveal 1:48:10 This Game's Winner Is... 1:53:18 Bonus Music/Outro

AUGUST ABSURDITY CONTINUES! David and Jessie renew their heated rivalry for a shot at the finals! Who will take one step closer to a return to glory!? Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:09:30 Game 1 0:14:16 Game 1 Reveal 0:30:31 Game 2 0:36:54 Game 2 Reveal 0:50:26 Game 3 0:54:33 Game 3 Reveal 1:11:22 Game 4 1:18:47 Game 4 Reveal 1:38:40 Game 5 1:45:53 Game 5 Reveal 1:59:18 Game 6! 2:04:14 Game 6 Reveal 2:19:29 This Game's Winner Is... 2:26:06 Bonus Music/Outro

AUGUST ABSURDITY RETURNS! Will Johnny be able to continue his ignominious reign as Absurd King?! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO!

It's time for a very special edition of the VGM Fight Club, the contest of VGM taste! In this episode, Jessie and David go head to head in a series of Olympic themed events to see who has the best taste in tunes and which countries will leave the Fight Club with the coveted gold medals! It's an international parade of fantastic music, enjoy! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Intro Racing 0:09:47 Richard Jacques (UK) - Sonic & All-Stars Racing Transformed - Burning Depths 0:14:29 Kazumi Totaka (JPN) - Wave Race 64 - Sandy Beach Judo 0:19:41 Keiichi Okabe (JPN) - Tekken 3 - Hwoarang (PlayStation2 Version) 0:23:26 Mick Gordon (AUS) - Killer Instinct - Touch Me and I'll Break Your Face Swimming 0:31:08 David Wise (UK) - Donkey Kong Country: Tropical Freeze - Amiss Abyss 0:35:17 Grant Kirkhope (SCO) - Banjo-Kazooie - Rusty Bucket Bay (Aquatic) Gymnastics 0:41:42 David Bergeaud (FRA) - Ratchet & Clank - Ride the Robot Train 0:44:08 Tim Follin (UK) - Treasure Master - Title Fencing 0:51:54 Tee Lopes (POR) - Teenage Mutant Ninja Turtles: Shredder's Revenge - Broadway Loop 0:55:39 Anamanaguchi (USA) - Scott Pilgrim vs. the World: The Game - Twin Dragons Freestyle 1:03:22 Toru Minegishi (JPN) - The Legend of Zelda: Twilight Princess - The Hidden Village 1:07:24 Jeff van Dyck (CAN) - Skitchin' - Jerky Curtains 1:19:38 Closing Ceremony: Koji Kondo (JPN) - Super Mario World - Staff Roll/Cast List

We're relaxing in the summer sun and enjoying incredibly hot VGM tunes! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:05:15 Ten the Hard Way! 0:17:32 Game 1 0:22:35 Game 1 Reveal 0:36:24 Game 2 0:40:33 Game 2 Reveal 0:53:42 Game 3 0:57:19 Game 3 Reveal 1:03:11 Lightning Round! 1:11:49 Game 4 1:18:18 Game 4 Reveal 1:32:00 VGM Threesome! 2:00:17 This Game's Winner Is... 2:08:47 Bonus Music/Outro

We've got memories, not all of them reliable or true, on this weeks episode! Join us in Discord to suggest new Lunar Gary clues and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:07:07 Game 1 0:12:13 Game 1 Reveal 0:29:23 Game 2 0:37:07 Game 2 Reveal 0:47:02 Game 3 0:57:47 Game 3 Reveal 1:15:59 Game 4 1:22:21 Game 4 Reveal 1:32:55 Game 5 1:39:42 Game 5 Reveal 1:50:57 This Game's Winner Is... 1:58:46 Bonus Music/Outro

We're feeling explosively sharp on this week's episode, because the show is all about FIREWORKS and BLADES! Enjoy this episode from a safe distance and with your parents supervision! Join us in Discord to suggest new Lunar Gary clues and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Sam L, PhoenixTear2121, BeastPond and HCO! 0:00:00 Welcome to Game That Tune! 0:08:05 Game 1 0:12:46 Game 1 Reveal 0:23:42 Game 2 0:29:29 Game 2 Reveal 0:45:02 Game 3 0:51:53 Game 3 Reveal 1:05:32 Game 4 1:14:01 Game 4 Reveal 1:26:52 Game 5 1:33:20 Game 5 Reveal 1:58:45 This Game's Winner Is... 2:07:29 Bonus Music/Outro

Jessie has put together a very special mixtape to celebrate Pride Month! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Taylor Y, Sam L, PhoenixTear2121, BeastPond, TheKerrigan and HCO! 0:00:00 Brat Overflow - 13 Sentinels: Aegis Rim 0:01:54 Intro 0:06:34 Every Day Is Night - VA-11 HALL-A 0:10:15 Asellus' Theme - Saga Frontier 0:13:18 Sub Battle - Bahamut Lagoon 0:14:52 Val Voyeaux - Dragon Age Inquisition 0:16:00 Eliminate the Impossible - Dream Daddy: A Dad Dating Simulator 0:17:38 Food Donkey - Night in the Woods 0:19:56 Song of Feathers (Emily's Theme) - Stardew Valley 0:22:43 Sunset Song - Paradise Killer 0:25:55 Dedication - Gone Home 0:26:57 Dynamite - Star Ocean Second Story 0:29:58 Ecstatic Vibrations, Totally Translucent - Disco Elysium 0:33:29 Simply the Best! - The Outer Worlds 0:34:38 Chandlo and Snorpy Head Home - Bugsnax 0:37:42 Wushu Dolls - Cyberpunk 2077 0:39:55 Helmut Uncorked - Psychonauts 2 0:41:08 Fallin' - Boyfriend Dungeon 0:44:45 Good Riddance - Hades

We're trying out some new games on this casual goofin' episode! Get ready for ten rounds of downright esoteric clues as John presents his vision of Lunar Gary, the guessing game of opposite titles! Then Jessie brings his all new Game That BOOM to the show for an audio quiz of classic gaming sound effects! Oh and we close with a discussion of Johnny's professional life, and whether or not 145 TB of pornography is too much for one free Google Drive account! Enjoy! Join us in Discord to suggest new Lunar Gary clues and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Taylor Y, Sam L, PhoenixTear2121, BeastPond, TheKerrigan and HCO!

We've got games inside of games in this week's show because we're playing MINIGAMES! We've got Johnny's sports power rankings, John's inability to feel a sense of urgency, David's boyhood dream come true, Jessie's goth girl fantasies, and most importantly, awesome VGM tunes! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Taylor Y, Sam L, PhoenixTear2121, BeastPond, TheKerrigan and HCO! 0:00:00 Welcome to Game That Tune! 0:06:19 Game 1 0:10:21 Game 1 Reveal 0:26:40 Game 2 0:33:02 Game 2 Reveal 0:50:26 Game 3 0:54:33 Game 3 Reveal 1:07:15 Game 4 1:13:16 Game 4 Reveal 1:27:02 Game 5 1:31:34 Game 5 Reveal 1:49:11 This Game's Winner Is... 1:57:04 Bonus Music/Outro

It's an all-new format for this week's GTT, but a format that VGM podcast fans may find familiar! We've taken this concept, along with the blessing of Darryl Bowers aka The Last Rican, and we're bringing you the first ever episode of the GTT VGM Fight Club! Johnny and Jessie are facing off in a battle of bangers to decide who has the greatest VGM taste and it's up to our live viewers on Twitch to decide! We hope you enjoy! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Taylor Y, Sam L, PhoenixTear2121, BeastPond, TheKerrigan and HCO! 0:00:00 Welcome to GTT VGM Fight Club! 0:08:13 Hypnospace Outlaw - I Am the Chowder Man 0:12:37 Super Lesbian Animal RPG - STAR ☆ POWER 0:22:51 Bloody Roar 3 - Dangerous Expectation - Prologue 0:26:43 Penny Time - Neon 0:35:16 Final Fantasy VIII - The Man With the Machine Gun 0:38:52 Foxy 2 - Main Theme 0:44:52 The Messenger - The Frozen Dark (Glacial Peak) 0:47:28 F-Zero X - Silence (Dream Chaser) 0:55:38 Sonic Mania - Chemical Plant Zone Boss ~ Mean Bean Machine 0:58:46 No More Heroes 2 - Philistine 1:15:50 Sky Shark - Stage 4

We've got games with cards on this week's episode, so be ready for open world adventures between minigames, wild character crossovers that happened and should've happened, gender non-conformity, and most importantly awesome VGM tunes! Join us in Discord and at GameThatTune.club! Check out our Patreon page! Patreon.com/GameThatTune is the home for exclusive content! Special thanks to our ABSURD FAN tier Patreon producers: Daniel Perkey, Taylor Y, Sam L, PhoenixTear2121, BeastPond, TheKerrigan and HCO! 0:00:00 Welcome to Game That Tune! 0:08:56 Game 1 0:16:40 Game 1 Reveal 0:29:46 Game 2 0:36:51 Game 2 Reveal 0:50:22 Game 3 0:56:50 Game 3 Reveal 1:10:05 Game 4 1:16:59 Game 4 Reveal 1:29:14 Game 5 1:34:00 Game 5 Reveal 1:46:59 This Game's Winner Is... 1:53:09 Bonus Music/Outro

OutlineChapter 14- Hypovolemic States- Etiology - True volume depletion occurs when fluid is lost from from the extracellular fluid at a rate exceeding intake - Can come the GI tract - Lungs - Urine - Sequestration in the body in a “third space” that is not in equilibrium with the extracellular fluid. - When losses occur two responses ameliorate them - Our intake of Na and fluid is way above basal needs - This is not the case with anorexia or vomiting - The kidney responds by minimizing further urinary losses - This adaptive response is why diuretics do not cause progressive volume depletion - Initial volume loss stimulates RAAS, and possibly other compensatory mechanisms, resulting increased proximal and collecting tubule Na reabsorption. - This balances the diuretic effect resulting in a new steady state in 1-2weeks - New steady state means Na in = Na out - GI Losses - Stomach, pancreas, GB, and intestines secretes 3-6 liters a day. - Almost all is reabsorbed with only loss of 100-200 ml in stool a day - Volume depletion can result from surgical drainage or failure of reabsorption - Acid base disturbances with GI losses - Stomach losses cause metabolic alkalosis - Intestinal, pancreatic and biliary secretions are alkalotic so losing them causes metabolic acidosis - Fistulas, laxative abuse, diarrhea, ostomies, tube drainage - High content of potassium so associated with hypokalemia - [This is a mistake for stomach losses] - Bleeding from the GI tract can also cause volume depletion - No electrolyte disorders from this unless lactic acidosis - Renal losses - 130-180 liters filtered every day - 98-99% reabsorbed - Urine output of 1-2 liters - A small 1-2% decrease in reabsorption can lead to 2-4 liter increase in Na and Water excretion - 4 liters of urine output is the goal of therapeutic diuresis which means a reduction of fluid reabsorption of only 2% - Diuretics - Osmotic diuretics - Severe hyperglycemia can contribute to a fluid deficit of 8-10 Iiters - CKD with GFR < 25 are poor Na conservers - Obligate sodium losses of 10 to 40 mEq/day - Normal people can reduce obligate Na losses down to 5 mEq/day - Usually not a problem because most people eat way more than 10-40 mEq of Na a day. - Salt wasting nephropathies - Water losses of 2 liters a day - 100 mEq of Na a day - Tubular and interstitial diseases - Medullary cystic kidney - Mechanism - Increased urea can be an osmotic diuretic - Damage to tubular epithelium can make it aldo resistant - Inability to shut off natriuretic hormone (ANP?) - The decreased nephro number means they need to be able to decrease sodium reabsorption per nephron. This may not be able to be shut down acutely. - Experiment, salt wasters can stay in balance if sodium intake is slowly decreased. (Think weeks) - Talks about post obstruction diuresis - Says it is usually appropriate rather than inappropriate physiology. - Usually catch up solute and water clearance after releasing obstruction - Recommends 50-75/hr of half normal saline - Talks briefly about DI - Skin and respiratory losses - 700-1000 ml of water lost daily by evaporation, insensible losses (not sweat) - Can rise to 1-2 liters per hour in dry hot climate - 30-50 mEq/L Na - Thirst is primary compensation for this - Sweat sodium losses can result in hypovolemia - Burns and exudative skin losses changes the nature of fluid losses resulting in fluid losses more similar to plasma with a variable amount of protein - Bronchorrhea - Sequestration into a third space - Volume Deficiency produced by the loss of interstitial and intravascular fluid into a third space that is not in equilibrium with the extracellular fluid. - Hip fracture 1500-2000 into tissues adjacent to fxr - Intestinal obstruction, severe pancreatitis, crush injury, bleeding, peritonitis, obstruction of a major venous system - Difference between 3rd space and cirrhosis ascities - Rate of accumulation, if the rate is slow enough there is time for renal sodium and water compensation to maintain balance. - So cirrhotics get edema from salt retension and do not act as hypovolemia - Hemodynamic response to volume depletion - Initial volume deficit reduced venous return to heart - Detected by cardiopulmonary receptors in atria and pulmonary veins leading to sympathetic vasoconstriction in skin and skeletal muscle. - More marked depletion will result in decreased cardiac output and decrease in BP - This drop in BP is now detected by carotid and aortic arch baroreceptors resulting in splanchnic and renal circulation vasoconstriction - This maintains cardiac and cerebral circulation - Returns BP toward normal - Increase in BP due to increased venous return - Increased cardiac contractility and heart rate - Increased vascular resistance - Sympathetic tone - Renin leading to Ang2 - These can compensate for 500 ml of blood loss (10%) - Unless there is autonomic dysfunction - With 16-25% loss this will not compensate for BP when patient upright - Postural dizziness - Symptoms - Three sets of symptoms can occur in hypovolemic patients - Those related to the manner in which the fluid loss occurs - Vomiting - Diarrhea - Polyuria - Those due to volume depletion - Those due to the electrode and acid base disorders that can accompany volume depletion - The symptoms of volume depletion are primarily related to the decrease in tissue perfusion - Early symptoms - Lassitude - Fatiguability - Thirst - Muscle cramps - Postural dizziness - As it gets more severe - Abdominal pain - Chest pain - Lethargy - Confusion - Symptomatic hypovolemia is most common with isosmotic Na and water depletion - In contrast pure water loss, causes hypernatremia, which results in movement of water from the intracellular compartment to the extracellular compartment, so that 2/3s of volume loss comes from the intracellular compartment, which minimizes the decrease in perfusion - Electrolyte disorders and symptoms - Muscle weakness from hypokalemia - Polyuria/poly dips is from hyperglycemia and hypokalemia - Lethargy, confusion, Seizures, coma from hyponatremia, hypernatremia, hyperglycemia - Extreme salt craving is unique to adrenal insufficiency - Eating salt off hands ref 18 - Evaluation of the hypovolemic patient - Know that if the losses are insensible then the sodium should rise - Volume depletion refers to extracellular volume depletion of any cause, while dehydration refers to the presence of hypernatremia due to pure water loss. Such patients are also hypovolemic. - Physical exam is insensitive and nonspecific - Finding most sensitive and specific finding for bleeding is postural changes in blood pressure - I don't find this very specific at all! - Recommends laboratory confirmation regardless of physical exam - Skin and mucous membranes - Should return too shape quickly - Elastic property is called Turgur - Not reliable is patients older than 55 to 60 - Dry axilla - Dry mucus membranes - Dark skin in Addison's disease Frim increased ACTH - Arterial BP - As volume goes down so does arterial BP - Marked fluid loss leads to quiet korotkoff signs - Interpret BP in terms of the patients “normal BP” - Venous pressure - Best done by looking at the JVP - Right atrial and left atrial pressure - LV EDP is RAP + 5 mmHg - Be careful if valvular disease, right heart failure, cor pulmonare, - Figure 14-2 - Shock - 30% blood loss - Lab Data - Urine Na concentration - Should be less than 25 mmol/L, can go as low as 1 mmol/L - Metabolic alkalosis can throw this off - Look to the urine chloride - Figure 14-3 - Renal artery stenosis can throw this off - FENa - Mentions that it doesn't work so well at high GFR - Urine osmolality - Indicates ADH - Volume depletion often associated with urine osm > 450 - Impaired by - Renal disease - Osmotic diuretic - Diuretics - DI - Mentions that severe volume depletion and hypokalemia impairs urea retension in renal medulla - Points out that isotonic urine does not rule out hypovolemia - Mentions specific gravity - BUN and Cr concentration - Normal ratio is 10:1 - Volume depletion this goes to 20:1 - Serum Na - Talks about diarrhea - Difference between secretory diarrhea which is isotonic and just causes hypovolemia - And osmotic which results in a lower electrolyte content and development of hypernatremia - Talks about hyperglycemia - Also can cause the sodium to rise from the low electrolyte content of the urine - But the pseudohyponatraemia can protect against this - Plasma potassium - Treatment - Both oral and IV treatment can be used for volume replacement - The goal of therapy are to restore normovolemia - And to correct associated acid-base and electrolyte disorders - Oral Therapy - Usually can be accomplished with increased water and dietary sodium - May use salt tablets - Glucose often added to resuscitation fluids - Provides calories - Promotes intestinal Na reabsorption since there is coupled Na and Glucose similar to that seen in the proximal tubule - Rice based solutions provide more calories and amino acids which also promote sodium reabsorption - 80g/L of glucose with rice vs 20 g/L with glucose alone - IV therapy - Dextrose solutions - Physiologically equivalent to water - For correcting hypernatremia - For covering insensible losses - Watch for hyperglycemia - Footnote warns against giving sterile water - Saline solutions - Most hypovolemic patients have a water and a sodium deficit - Isotonic saline has a Na concentration of 154, similar to that of plasma see page 000 - Half-isotonic saline is equivalent to 550 ml of isotonic saline and 500 of free water. Is that a typo? - 3% is a liter of hypertonic saline and 359 extra mEq of Na - Dextrose in saline solutions - Give a small amount of calories, otherwise useless - Alkalinizing solutions - 7.5% NaHCO3 in 50 ml ampules 44 mEq of Na and 44 mEq of HCO3 - Treat metabolic acidosis or hyperkalemia - Why 44 mEq and not 50? - Do not give with calcium will form insoluble CaCO3 - Polyionic solutions - Ringers contains physiologic K and Ca - Lactated Ringers adds 28 mEq of lactate - Spreads myth of LR in lactic acidosis - Potassium chloride - Available as 2 mEq/mL - Do not give as a bolus as it can cause fatal hyperkalemia - Plasma volume expanders - Albumin, polygelastins, hetastarch are restricted to vascular space - 25% albumin can pull fluid into the vascular space - 25% albumin is an albumin concentration of 25 g/dL compare to physiologic 4 g/dL - Says it pulls in several times its own volume - 5% albumin is like giving plasma - Blood - Which fluid? - Look at osmolality, give hypotonic fluids to people with high osmolality - Must include all electrolytes - Example of adding 77 mEw of K to 0.45 NS and making it isotonic - DI can be replaced with dextrose solutions, pure water deficit - Case 14-3 - Diarrhea with metabolic acidosis - He chooses 0.25 NS with 44 mEq of NaCl and 44 NaHCO3 - Talks about blood and trauma - Some studies advocate delaying saline until penetrating trauma is corrected APR about to. Keep BP low to prevent bleeding. Worry about diluting coagulation factors - Only do this if the OR is quickly available - Volume deficit - Provides formula for water deficit and sodium deficit - Do not work for isotonic losses - Provides a table to adjust fluid loss based on changes in Hgb or HCTZ - Says difficult to estimate it from lab findings and calculations - Follow serial exams - Serial urine Na - Rate of replacement - Goal is not to give fluid but to induce a positive balance - Suggests 50-100 ml/hr over what is coming out of the body - Urine - Insensibles 30-50 - Diarrhea - Tubes - Hypovolemic shock - Due to bleeding - Sequesting in third space - Why shock? - Progressive volume depletion leads to - Increased sympathetic NS - Increased Ang 2 - Initially this maintains BP, cerebral and coronary circulation - But this can decrease splanchnic, renal and mucocutaneous perfusion - This leads to lactic acicosis - This can result in intracellular contents moving into circulation or translocation of gut bacteria - Early therapy to prevent irreversible shock - In dogs need to treat with in 2 hours - In humans may need more than 4 hours - Irreversible shock associated with pooling of blood in capillaries - Vasomotor paralysis - Hyperpolarization of vascular smooth muscle as depletion of ATP allows K to flowing out from K channels opening. Ca flows out too leading to vasodilation - Glyburide is an K-ATP channel inhibitor (?) caused increased vasoconstriction and BP - Pluggin of capillaries by neutrophils - Cerebral ischemia - Increased NO generation - Which Fluids? - Think of what is lost and replace that. - Bleeding think blood - Raise the hct but not above 35 - Acellular blood substitutes, looked bad at the time of this writing - Di aspirin cross linked hemoglobin had increased 2 and 28 day mortality vs saline - Colloids sound great but they fail in RCTs - SAFE - FEAST - Points out that saline replaces the interstitial losses why do we think those losses are unimportant - Pulmonary circulation issue - Pulmonary circulation is more leaky so oncotic pressure less effective there - Talks about the lungs be naturally protected from pulmonary edema - Rate of fluid - 1-2 liters in first hour - Suggests CVP or capillary wedge pressure during resuscitation - No refs in the rate of fluid administration section - Lactic acidosis - Points out that HCO can impair lactate utilization - Also states that arterial pH does not point out what is happening at the tissue level. Suggests mixed-venous sample.ReferencesJCI - Phenotypic and pharmacogenetic evaluation of patients with thiazide-induced hyponatremia and a nice review of this topic: Altered Prostaglandin Signaling as a Cause of Thiazide-Induced HyponatremiaThe electrolyte concentration of human gastric secretion. https://physoc.onlinelibrary.wiley.com/doi/10.1113/expphysiol.1960.sp001428A classic by Danovitch and Bricker: Reversibility of the “Salt-Losing” Tendency of Chronic Renal Failure | NEJMOsmotic Diuresis Due to Retained Urea after Release of Obstructive Uropathy | NEJMIs This Patient Hypovolemic? | Cardiology | JAMAAnd by the same author, a textbook: Steven McGee. 5th edition. Evidence-Based Physical Diagnosis Elsevier Philadelphia 2022. ISBN-13: 978-0323754835The clinical course and pathophysiological investigation of adolescent gestational diabetes insipidus: a case report | BMC Endocrine DisordersSensitivity and specificity of clinical signs for assessment of dehydration in endurance athletes | British Journal of Sports MedicineDiagnostic performance of serum blood urea nitrogen to creatinine ratio for distinguishing prerenal from intrinsic acute kidney injury in the emergency department | BMC NephrologyThe meaning of the blood urea nitrogen/creatinine ratio in acute kidney injury - PMCLanguage guiding therapy: the case for dehydration vs volume depletion https://www.acpjournals.org/doi/10.7326/0003-4819-127-9-199711010-00020?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub%20%200pubmedValidation of a noninvasive monitor to continuously trend individual responses to hypovolemiaReferences for Anna's voice of God on Third Spacing : Shires Paper from 1964 (The ‘third space' – fact or fiction? )References for melanie's VOG:1. Appraising the Preclinical Evidence of the Role of the Renin-Angiotensin-Aldosterone System in Antenatal Programming of Maternal and Offspring Cardiovascular Health Across the Life Course: Moving the Field Forward: A Scientific Statement From the American Heart Association2. excellent review of RAAS in pregnancy: The enigma of continual plasma volume expansion in pregnancy: critical role of the renin-angiotensin-aldosterone systemhttps://journals-physiology-org.ezp-prod1.hul.harvard.edu/doi/full/10.1152/ajprenal.00129.20163. 10.1172/JCI107462- classic study in JCI of AngII responsiveness during pregnancy4. William's Obstetrics 26th edition!5. Feto-maternal osmotic balance at term. A prospective observational study

¿Quieres entrenar con nosotros?. Guiado y de la mano. Es aquí https://bit.ly/quiero_entrenar ------------------------------------------------- Charlamos con Aitor Viribay, responsable del área de rendimiento del equipo INEOS Granadiers (equipo ciclista de talla World Tour). Hablamos de: - Los pros y contras de trabajar con algunos de los mejores ciclistas del mundo - Ingerir 120 gramos de HCO/hora durante el ejercicio. ¿Qué dice la ciencia? ¿Es para todo el mundo? - Errores habituales que cometen los deportistas amateurs en su nutrición. - Consejos para optimizar la absorción de nutrientes durante el ejercicio - Flexibilidad metabólica ¿Cómo influye? - Recomendaciones de alimentación durante el ejercicio - Dietas Low Carb - Aprendizajes de Aitor tras trabajar con Killian Jornet Web divulgativa de Aitor: https://glut4science.com/ Redes sociales de Aitor https://www.instagram.com/aitorviribaymorales?igsh=MTRqbnZvN3l0OHR6dw==

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.28.550873v1?rss=1 Authors: Glass, M. R., Waxman, E. A., Yamashita, S., Lafferty, M., Beltran, A., Farah, T., Patel, N. K., Matoba, N., Ahmed, S., Srivastava, M., Drake, E., Davis, L. T., Yeturi, M., Sun, K., Love, M. I., Hashimoto-Torii, K., French, D. L., Stein, J. L. Abstract: Background: Reproducibility of hCO phenotypes remains a concern for modeling neurodevelopmental disorders. While guided human cortical organoid (hCO) protocols reproducibly generate cortical cell types in multiple cell lines at one site, variability across sites using a harmonized protocol has not yet been evaluated. We present an hCO cross-site reproducibility study examining multiple phenotypes. Methods: Three independent research groups generated hCOs from one induced pluripotent stem cell (iPSC) line using a harmonized miniaturized spinning bioreactor protocol. scRNA-seq, 3D fluorescent imaging, phase contrast imaging, qPCR, and flow cytometry were used to characterize the 3 month differentiations across sites. Results: In all sites, hCOs were mostly cortical progenitor and neuronal cell types in reproducible proportions with moderate to high fidelity to the in vivo brain that were consistently organized in cortical wall-like buds. Cross-site differences were detected in hCO size and morphology. Differential gene expression showed differences in metabolism and cellular stress across sites. Although iPSC culture conditions were consistent and iPSCs remained undifferentiated, primed stem cell marker expression prior to differentiation correlated with cell type proportions in hCOs. Conclusions: We identified hCO phenotypes that are reproducible across sites using a harmonized differentiation protocol. Previously described limitations of hCO models were also reproduced including off-target differentiations, necrotic cores, and cellular stress. Improving our understanding of how stem cell states influence early hCO cell types may increase reliability of hCO differentiations. Cross-site reproducibility of hCO cell type proportions and organization lays the foundation for future collaborative prospective meta-analytic studies modeling neurodevelopmental disorders in hCOs. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

ReferencesWe considered the complexity of the machinery to excrete ammonium in the context of research on dietary protein and how high protein intake may increase glomerular pressure and contribute to progressive renal disease (many refer to this as the “Brenner hypothesis”). Dietary protein intake and the progressive nature of kidney disease: the role of hemodynamically mediated glomerular injury in the pathogenesis of progressive glomerular sclerosis in aging, renal ablation, and intrinsic renal diseaseA trial that studied low protein and progression of CKD The Effects of Dietary Protein Restriction and Blood-Pressure Control on the Progression of Chronic Renal Disease(and famously provided data for the MDRD eGFR equation A more accurate method to estimate glomerular filtration rate from serum creatinine: a new prediction equation. Modification of Diet in Renal Disease Study GroupWe wondered about dietary recommendations in CKD. of note, this is best done in the DKD guidelines from KDIGO Executive summary of the 2020 KDIGO Diabetes Management in CKD Guideline: evidence-based advances in monitoring and treatment.Joel mentioned this study on red meat and risk of ESKD. Red Meat Intake and Risk of ESRDWe referenced the notion of a plant-based diet. This is an excellent review by Deborah Clegg and Kathleen Hill Gallant. Plant-Based Diets in CKD : Clinical Journal of the American Society of NephrologyHere's the review that Josh mentioned on how the kidney appears to sense pH Molecular mechanisms of acid-base sensing by the kidneyRemarkably, Dr. Dale Dubin put a prize in his ECG book Free Car Prize Hidden in Textbook Read the fine print: Student wins T-birdA review of the role of the kidney in DKA: Diabetic ketoacidosis: Role of the kidney in the acid-base homeostasis re-evaluatedJosh mentioned the effects of infusing large amounts of bicarbonate The effect of prolonged administration of large doses of sodium bicarbonate in man and this study on the respiratory response to a bicarbonate infusion: The Acute Effects In Man Of A Rapid Intravenous Infusion Of Hypertonic Sodium Bicarbonate Solution. Ii. Changes In Respiration And Output Of Carbon DioxideThis is the study of acute respiratory alkalosis in dogs: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC293311/?page=1And this is the study of medical students who went to the High Alpine Research Station on the Jungfraujoch in the Swiss Alps https://www.nejm.org/doi/full/10.1056/nejm199105163242003Self explanatory! A group favorite! It Is Chloride Depletion Alkalosis, Not Contraction AlkalosisEffects of chloride and extracellular fluid volume on bicarbonate reabsorption along the nephron in metabolic alkalosis in the rat. Reassessment of the classical hypothesis of the pathogenesis of metabolic alkalosisA review of pendrin's role in volume homeostasis: The role of pendrin in blood pressure regulation | American Journal of Physiology-Renal PhysiologyInfusion of bicarbonate may lead to a decrease in respiratory stimulation but the shift of bicarbonate to the CSF may lag. Check out this review Neural Control of Breathing and CO2 Homeostasis and this classic paper Spinal-Fluid pH and Neurologic Symptoms in Systemic Acidosis.OutlineOutline: Chapter 11- Regulation of Acid-Base Balance- Introduction - Bicarb plus a proton in equilibrium with CO2 and water - Can be rearranged to HH - Importance of regulating pCO2 and HCO3 outside of this equation - Metabolism of carbs and fats results in the production of 15,000 mmol of CO2 per day - Metabolism of protein and other “substances” generates non-carbonic acids and bases - Mostly from sulfur containing methionine and cysteine - And cationic arginine and lysine - Hydrolysis of dietary phosphate that exists and H2PO4– - Source of base/alkali - Metabolism of an ionic amino acids - Glutamate and asparatate - Organic anions going through gluconeogenesis - Glutamate, Citrate and lactate - Net effect on a normal western diet 50-100 mEq of H+ per day - Homeostatic response to these acid-base loads has three stages: - Chemical buffering - Changes in ventilation - Changes in H+ excretion - Example of H2SO4 from oxidation of sulfur containing AA - Drop in bicarb will stimulate renal acid secretion - Nice table of normal cid-base values, arterial and venous- Great 6 bullet points of acid-base on page 328 - Kidneys must excrete 50-100 of non-carbonic acid daily - This occurs by H secretion, but mechanisms change by area of nephron - Not excreted as free H+ due to minimal urine pH being equivalent to 0.05 mmol/L - No H+ can be excreted until virtually all of th filtered bicarb is reabsorbed - Secreted H+ must bind buffers (phosphate, NH3, cr) - PH is main stimulus for H secretion, though K, aldo and volume can affect this.- Renal Hydrogen excretion - Critical to understand that loss of bicarb is like addition of hydrogen to the body - So all bicarb must be reabsorbed before dietary H load can be secreted - GFR of 125 and bicarb of 24 results in 4300 mEq of bicarb to be reabsorbed daily - Reabsorption of bicarb and secretion of H involve H secretion from tubular cells into the lumen. - Thee initial points need to be emphasized - Secreted H+ ion are generated from dissociation of H2O - Also creates OH ion - Which combine with CO2 to form HCO3 with the help of zinc containing intracellular carbonic anhydrase. - This is how the secretion of H+ which creates an OH ultimately produces HCO3 - Different mechanisms for proximal and distal acidification - NET ACID EXCRETION - Free H+ is negligible - So net H+ is TA + NH4 – HCO3 loss - Unusually equal to net H+ load, 50-100 mEq/day - Can bump up to 300 mEq/day if acid production is increased - Net acid excretion can go negative following a bicarb or citrate load - Proximal Acidification - Na-H antiporter (or exchanger) in luminal membrane - Basolateral membrane has a 3 HCO3 Na cotransporter - This is electrogenic with 3 anions going out and only one cation - The Na-H antiporter also works in the thick ascending limb of LOH - How about this, there is also a H-ATPase just like found in the intercalated cells in the proximal tubule and is responsible for about a third of H secretion - And similarly there is also. HCO3 Cl exchanger (pendrin-like) in the proximal tubule - Footnote says the Na- 3HCO3 cotransporter (which moves sodium against chemical gradient NS uses negative charge inside cell to power it) is important for sensing acid-base changes in the cell. - Distal acidification - Occurs in intercalated cells of of cortical and medullary collecting tubule - Three main characteristics - H secretion via active secretory pumps in the luminal membrane - Both H-ATPase and H-K ATPase - H- K ATPase is an exchange pump, k reabsorption - H-K exchange may be more important in hypokalemia rather than in acid-base balance - Whole paragraph on how a Na-H exchanger couldn't work because the gradient that H has to be pumped up is too big. - H-ATPase work like vasopressin with premise H-ATPase sitting on endocarditis vesicles a=which are then inserted into the membrane. Alkalosis causes them to be recycled out of the membrane. - H secretory cells do not transport Na since they have few luminal Na channels, but are assisted by the lumen negative tubule from eNaC. - Minimizes back diffusion of H+ and promotes bicarb resorption - Bicarbonate leaves the cell through HCO3-Cl exchanger which uses the low intracellular Cl concentration to power this process. - Same molecule is found on RBC where it is called band 3 protein - Figure 11-5 is interesting - Bicarbonate resorption - 90% in the first 1-22 mm of the proximal tubule (how long is the proximal tubule?) - Lots of Na-H exchangers and I handed permeability to HCO3 (permeability where?) - Last 10% happens distally mostly TAL LOH via Na-H exchange - And the last little bit int he outer medullary collecting duct. - Carbonic anhydrase and disequilibrium pH - CA plays central role in HCO3 reabsorption - After H is secreted in the proximal tubule it combines with HCO# to form carbonic acid. CA then dehydrates it to CO2 and H2O. (Step 2) - Constantly moving carbonic acid to CO2 and H2O keeps hydrogen combining with HCO3 since the product is rapidly consumed. - This can be demonstrated by the minimal fall in luminal pH - That is important so there is not a luminal gradient for H to overcome in the Na-H exchanger (this is why we need a H-ATPase later) - CA inhibitors that are limited tot he extracellular compartment can impair HCO3 reabsorption by 80%. - CA is found in S1, S2 but not S3 segment. See consequence in figure 11-6. - The disequilibrium comes from areas where there is no CA, the HH formula falls down because one of the assumptions of that formula is that H2CO3 (carbonic acid) is a transient actor, but without CA it is not and can accumulate, so the pKa is not 6.1. - Bicarbonate secretion - Type B intercalated cells - H-ATPase polarity reversed - HCO3 Cl exchanger faces the apical rather than basolateral membrane- Titratable acidity - Weak acids are filtered at the glom and act as buffers in the urine. - HPO4 has PKA of 6.8 making it ideal - Creatinine (pKa 4.97) and uric acid (pKa 5.75) also contribute - Under normal cinditions TA buffers 10-40 mEa of H per day - Does an example of HPO4(2-):H2PO4 (1-) which exists 4:1 at pH of 7.4 (glomerular filtrate) - So for 50 mEq of Phos 40 is HPO4 and 10 is H2PO4 - When pH drops to 6.8 then the ratio is 1:1 so for 50 - So the 50 mEq is 25 and 25, so this buffered an additional 15 mEq of H while the free H+ concentration increased from 40 to 160 nanomol/L so over 99.99% of secreted H was buffered - When pH drops to 4.8 ratio is 1:100 so almost all 50 mEq of phos is H2PO4 and 39.5 mEq of H are buffered. - Acid loading decreases phosphate reabsorption so more is there to act as TA. - Decreases activity of Na-phosphate cotransporter - DKA provides a novel weak acid/buffer beta-hydroxybutyrate (pKa 4.8) which buffers significant amount of acid (50 mEq/d).- Ammonium Excretion - Ability to excrete H+ as ammonium ions adds an important amount of flexibility to renal acid-base regulation - NH3 and NH4 production and excretion can be varied according to physiologic need. - Starts with NH3 production in tubular cells - NH3, since it is neutral then diffuses into the tubule where it is acidified by the low pH to NH4+ - NH4+ is ionized and cannot cross back into the tubule cells(it is trapped in the tubular fluid) - This is important for it acting as an important buffer eve though the pKa is 9.0 - At pH of 6.0 the ratio of NH3 to NH4 is 1:1000 - As the neutral NH3 is converted to NH4 more NH3 from theintracellular compartment flows into the tubular fluid replacing the lost NH3. Rinse wash repeat. - This is an over simplification and that there are threemajor steps - NH4 is produced in early proximal tubular cells - Luminal NH4 is partially reabsorbed in the TAL and theNH3 is then recycled within the renal medulla - The medullary interstitial NH3 reaches highconcentrations that allow NH3 to diffuse into the tubular lumen in the medullary collecting tubule where it is trapped as NH4 by secreted H+ - NH4 production from Glutamine which converts to NH4 and glutamate - Glutamate is converted to alpha-ketoglutarate - Alpha ketoglutarate is converted to 2 HCO3 ions - HCO3 sent to systemic circulation by Na-3 HCO3 transporter - NH4 then secreted via Na-H exchanger into the lumen - NH4 is then reabsorbed by NaK2Cl transporter in TAL - NH4 substitutes for K - Once reabsorbed the higher intracellular pH causes NH4 to convert to NH3 and the H that is removed is secreted through Na-H exchanger to scavenge the last of the filtered bicarb. - NH3 diffuses out of the tubular cells into the interstitium - NH4 reabsorption in the TAL is suppressed by hyperkalemia and stimulated by chronic metabolic acidosis - NH4 recycling promotes acid clearance - The collecting tubule has a very low NH3 concentration - This promotes diffusion of NH3 into the collecting duct - NH3 that goes there is rapidly converted to NH4 allowing more NH3 to diffuse in. - Response to changes in pH - Increased ammonium excretion with two processes - Increased proximal NH4 production - This is delayed 24 hours to 2-3 days depending on which enzyme you look at - Decreased urine pH increases diffusion of ammonia into the MCD - Occurs with in hours of an acid load - Peak ammonium excretion takes 5-6 days! (Fig 11-10) - Glutamine is picked up from tubular fluid but with acidosis get Na dependent peritublar capillary glutamine scavenging too - Glutamine metabolism is pH dependent with increase with academia and decrease with alkalemia - NH4 excretion can go from 30-40 mEq/day to > 300 with severe metabolic acidosis (38 NaBicarb tabs) - Says each NH4 produces equimolar generation of HCO3 but I thought it was two bicarb for every alpha ketoglutarate?- The importance of urine pH - Though the total amount of hydrogren cleared by urine pH is insignificant, an acidic urine pH is essential for driving the reactions of TA and NH4 forward.- Regulation of renal hydrogen excretion - Net acid excretion vary inverse with extracellular pH - Academia triggers proximal and distal acidification - Proximally this: - Increased Na-H exchange - Increased luminal H-ATPase activity - Increased Na:3HCO3 cotransporter on the basolateral membrane - Increased NH4 production from glutamine - In the collecting tubules - Increased H-ATPase - Reduction of tubular pH promotes diffusion of NH3 which gets converted to NH4…ION TRAPPING - Extracellular pH affects net acid excretion through its affect on intracellular pH - This happens directly with respiratory disorders due to movement of CO2 through the lipid bilayer - In metabolic disorders a low extracellular bicarb with cause bicarb to diffuse out of the cell passively, this lowers intracellular pH - If you manipulate both low pCO2 and low Bicarb to keep pH stable there will be no change in the intracellular pH and there is no change in renal handling of acid. It is intracellular pH dependent - Metabolic acidosis - Ramps up net acid secretion - Starts within 24 hours and peaks after 5-6 days - Increase net secretion comes from NH4 - Phosphate is generally limited by diet - in DKA titratable acid can be ramped up - Metabolic alkalosis - Alkaline extracellular pH - Increased bicarb excretion - Decrease reabsorption - HCO3 secretion (pendrin) in cortical collecting tubule - Occurs in cortical intercalated cells able to insert H-ATPase in basolateral cells (rather than luminal membrane) - Normal subjects are able to secrete 1000 mmol/day of bicarb - Maintenance of metabolic alkalosis requires a defect which forces the renal resorption of bicarb - This can be chloride/volume deficiency - Hypokalemia - Hyperaldosteronism - Respiratory acidosis and alkalosis - PCO2 via its effect on intracellular pH is an important determinant of renal acid handling - Ratios he uses: - 3.5 per 10 for respiratory acidosis - 5 per 10 for respiratory alkalosis - Interesting paragraph contrasting the response to chronic metabolic acidosis vs chronic respiratory acidosis - Less urinary ammonium in respiratory acidosis - Major differences in proximal tubule cell pH - In metabolic acidosis there is decreased bicarb load so less to be reabsorbed proximally - In respiratory acidosis the increased serum bicarb increases the amount of bicarb that must be reabsorbed proximally - The increased activity of Na-H antiporter returns tubular cell pH to normal and prevents it from creating increased urinary ammonium - Mentions that weirdly more mRNA for H-Na antiporter in metabolic acidosis than in respiratory acidosis - Net hydrogen excretion varies with effective circulating volume - Starts with bicarb infusions - Normally Tm at 26 - But if you volume deplete the patient with diuretics first this increases to 35+ - Four factors explain this increased Tm for bicarb with volume deficiency - Reduced GFR - Activation of RAAS - Ang2 stim H-Na antiporter proximally - Ang2 also stimulates Na-3HCO3 cotransporter on basolateral membrane - Aldosterone stimulates H-ATPase in distal nephron - ALdo stimulates Cl HCO3 exchanger on basolateral membrane - Aldo stimulates eNaC producing tubular lumen negative charge to allow H secretion to occur and prevents back diffusion - Hypochloremia - Increases H secretion by both Na-dependent and Na-independent methods - If Na is 140 and Cl is 115, only 115 of Na can be reabsorbed as NaCl, the remainder must be reabsorbed with HCO3 or associated with secretion of K or H to maintained electro neutrality - This is enhanced with hypochloridemia - Concurrent hypokalemia - Changes in K lead to trans cellular shifts that affect inctracellular pH - Hypokalemia causes K out, H in and in the tubular cell the cell acts if there is systemic acidosis and increases H secretion (and bicarbonate resorption) - PTH - Decreases proximal HCO3 resorption - Primary HyperCard as cause of type 2 RTA - Does acidosis stim PTH or does PTH stim net acid excretion

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.13.536491v1?rss=1 Authors: Stanton, M. M., Hariani, H. N., Sorokin, J., Taylor, P. M., Modan, S., Rash, B. G., Rao, S. B., Enriquez, L., Quang, D., Hsu, P.-K., Paek, J., Owango, D., Castrillo, C., Nicola, J., Ramkumar, P., Lash, A., Flanzer, D., Shah, K., Kato, S., Skibinski, G. Abstract: Human-derived cortical organoids (hCOs) recapitulate cell diversity and 3D structure found in the human brain and offer a promising model for discovery of new gene therapies targeting neurological disorders. Adeno-associated viruses (AAVs) are the most promising vehicles for non-invasive gene delivery to the central nervous system (CNS), but reliable and reproducible in vitro models to assess their clinical potential are lacking. hCOs can take on these issues as they are a physiologically relevant model to assess AAV transduction efficiency, cellular tropism, and biodistribution within the tissue parenchyma, all of which could significantly modulate therapeutic efficacy. Here, we examine a variety of naturally occurring AAV serotypes and measure their ability to transduce neurons and glia in hCOs from multiple donors. We demonstrate cell tropism driven by AAV serotype and hCO donor and quantify fractions of neurons and astrocytes transduced with GFP as well as overall hCO health. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

ReferencesWe considered the effect of a high protein diet and potential metabolic acidosis on kidney function. This review is of interest by Donald Wesson, a champion for addressing this issue and limiting animal protein: Mechanisms of Metabolic Acidosis-Induced Kidney Injury in Chronic Kidney DiseaseHostetter explored the effect of a high protein diet in the remnant kidney model with 1 ¾ nephrectomy. Rats with reduced dietary acid load (by bicarbonate supplementation) had less tubular damage. Chronic effects of dietary protein in the rat with intact and reduced renal massWesson explored treatment of metabolic acidosis in humans with stage 3 CKD in this study. Treatment of metabolic acidosis in patients with stage 3 chronic kidney disease with fruits and vegetables or oral bicarbonate reduces urine angiotensinogen and preserves glomerular filtration rateIn addition to the effect of metabolic acidosis from a diet high in animal protein, this diet also leads to hyperfiltration. This was demonstrated in normal subjects; ingesting a protein diet had a significantly higher creatinine clearance than a comparable group of normal subjects ingesting a vegetarian diet. Renal functional reserve in humans: Effect of protein intake on glomerular filtration rate.This finding has been implicated in Brenner's theory regarding hyperfiltration: The hyperfiltration theory: a paradigm shift in nephrologyOne of multiple publications from Dr. Nimrat Goraya whom Joel mentioned in the voice over: Dietary Protein as Kidney Protection: Quality or Quantity?We wondered about the time course in buffering a high protein meal (and its subsequent acid load on ventilation) and Amy found this report:Effect of Protein Intake on Ventilatory Drive | Anesthesiology | American Society of Anesthesiologists Roger mentioned that the need for acetate to balance the acid from amino acids in parenteral nutrition was identified in pediatrics perhaps because infants may have reduced ability to generate acid. Randomised controlled trial of acetate in preterm neonates receiving parenteral nutrition - PMCHe also recommended an excellent review on the complications of parenteral nutrition by Knochel https://www.kidney-international.org/action/showPdf?pii=S0085-2538%2815%2933384-6 which explained that when the infused amino acids disproportionately include cationic amino acids, metabolism led to H+ production. This is typically mitigated by preparing a solution that is balanced by acetate. Amy mentioned this study that explored the effect of protein intake on ventilation: Effect of Protein Intake on Ventilatory Drive | Anesthesiology | American Society of AnesthesiologistsAnna and Amy reminisced about a Skeleton Key Group Case from the renal fellow network Skeleton Key Group: Electrolyte Case #7JC wondered about isolated defects in the proximal tubule and an example is found here: Mutations in SLC4A4 cause permanent isolated proximal renal tubular acidosis with ocular abnormalitiesAnna's Voiceover re: Gastric neobladder → metabolic alkalosis and yes, dysuria. The physiology of gastrocystoplasty: once a stomach, always a stomach but not as common as you might think Gastrocystoplasty: long-term complications in 22 patientsSjögren's syndrome has been associated with acquired distal RTA and in some cases, an absence of the H+ ATPase, presumably from autoantibodies to this transporter. Here's a case report: Absence of H(+)-ATPase in cortical collecting tubules of a patient with Sjogren's syndrome and distal renal tubular acidosisCan't get enough disequilibrium pH? Check this out- Spontaneous luminal disequilibrium pH in S3 proximal tubules. Role in ammonia and bicarbonate transport.Acetazolamide secretion was studied in this report Concentration-dependent tubular secretion of acetazolamide and its inhibition by salicylic acid in the isolated perfused rat kidney. | Drug Metabolism & DispositionIn this excellent review, David Goldfarb tackles the challenging case of a A Woman with Recurrent Calcium Phosphate Kidney Stones (spoiler alert, many of these patients have incomplete distal RTA and this problem is hard to treat). Molecular mechanisms of renal ammonia transport excellent review from David Winer and Lee Hamm. OutlineOutline: Chapter 11- Regulation of Acid-Base Balance- Introduction - Bicarb plus a proton in equilibrium with CO2 and water - Can be rearranged to HH - Importance of regulating pCO2 and HCO3 outside of this equation - Metabolism of carbs and fats results in the production of 15,000 mmol of CO2 per day - Metabolism of protein and other “substances” generates non-carbonic acids and bases - Mostly from sulfur containing methionine and cysteine - And cationic arginine and lysine - Hydrolysis of dietary phosphate that exists and H2PO4– - Source of base/alkali - Metabolism of an ionic amino acids - Glutamate and asparatate - Organic anions going through gluconeogenesis - Glutamate, Citrate and lactate - Net effect on a normal western diet 50-100 mEq of H+ per day - Homeostatic response to these acid-base loads has three stages: - Chemical buffering - Changes in ventilation - Changes in H+ excretion - Example of H2SO4 from oxidation of sulfur containing AA - Drop in bicarb will stimulate renal acid secretion - Nice table of normal cid-base values, arterial and venous- Great 6 bullet points of acid-base on page 328 - Kidneys must excrete 50-100 of non-carbonic acid daily - This occurs by H secretion, but mechanisms change by area of nephron - Not excreted as free H+ due to minimal urine pH being equivalent to 0.05 mmol/L - No H+ can be excreted until virtually all of th filtered bicarb is reabsorbed - Secreted H+ must bind buffers (phosphate, NH3, cr) - PH is main stimulus for H secretion, though K, aldo and volume can affect this.- Renal Hydrogen excretion - Critical to understand that loss of bicarb is like addition of hydrogen to the body - So all bicarb must be reabsorbed before dietary H load can be secreted - GFR of 125 and bicarb of 24 results in 4300 mEq of bicarb to be reabsorbed daily - Reabsorption of bicarb and secretion of H involve H secretion from tubular cells into the lumen. - Thee initial points need to be emphasized - Secreted H+ ion are generated from dissociation of H2O - Also creates OH ion - Which combine with CO2 to form HCO3 with the help of zinc containing intracellular carbonic anhydrase. - This is how the secretion of H+ which creates an OH ultimately produces HCO3 - Different mechanisms for proximal and distal acidification - NET ACID EXCRETION - Free H+ is negligible - So net H+ is TA + NH4 – HCO3 loss - Unusually equal to net H+ load, 50-100 mEq/day - Can bump up to 300 mEq/day if acid production is increased - Net acid excretion can go negative following a bicarb or citrate load - Proximal Acidification - Na-H antiporter (or exchanger) in luminal membrane - Basolateral membrane has a 3 HCO3 Na cotransporter - This is electrogenic with 3 anions going out and only one cation - The Na-H antiporter also works in the thick ascending limb of LOH - How about this, there is also a H-ATPase just like found in the intercalated cells in the proximal tubule and is responsible for about a third of H secretion - And similarly there is also. HCO3 Cl exchanger (pendrin-like) in the proximal tubule - Footnote says the Na- 3HCO3 cotransporter (which moves sodium against chemical gradient NS uses negative charge inside cell to power it) is important for sensing acid-base changes in the cell. - Distal acidification - Occurs in intercalated cells of of cortical and medullary collecting tubule - Three main characteristics - H secretion via active secretory pumps in the luminal membrane - Both H-ATPase and H-K ATPase - H- K ATPase is an exchange pump, k reabsorption - H-K exchange may be more important in hypokalemia rather than in acid-base balance - Whole paragraph on how a Na-H exchanger couldn't work because the gradient that H has to be pumped up is too big. - H-ATPase work like vasopressin with premise H-ATPase sitting on endocarditis vesicles a=which are then inserted into the membrane. Alkalosis causes them to be recycled out of the membrane. - H secretory cells do not transport Na since they have few luminal Na channels, but are assisted by the lumen negative tubule from eNaC. - Minimizes back diffusion of H+ and promotes bicarb resorption - Bicarbonate leaves the cell through HCO3-Cl exchanger which uses the low intracellular Cl concentration to power this process. - Same molecule is found on RBC where it is called band 3 protein - Figure 11-5 is interesting - Bicarbonate resorption - 90% in the first 1-22 mm of the proximal tubule (how long is the proximal tubule?) - Lots of Na-H exchangers and I handed permeability to HCO3 (permeability where?) - Last 10% happens distally mostly TAL LOH via Na-H exchange - And the last little bit int he outer medullary collecting duct. - Carbonic anhydrase and disequilibrium pH - CA plays central role in HCO3 reabsorption - After H is secreted in the proximal tubule it combines with HCO# to form carbonic acid. CA then dehydrates it to CO2 and H2O. (Step 2) - Constantly moving carbonic acid to CO2 and H2O keeps hydrogen combining with HCO3 since the product is rapidly consumed. - This can be demonstrated by the minimal fall in luminal pH - That is important so there is not a luminal gradient for H to overcome in the Na-H exchanger (this is why we need a H-ATPase later) - CA inhibitors that are limited tot he extracellular compartment can impair HCO3 reabsorption by 80%. - CA is found in S1, S2 but not S3 segment. See consequence in figure 11-6. - The disequilibrium comes from areas where there is no CA, the HH formula falls down because one of the assumptions of that formula is that H2CO3 (carbonic acid) is a transient actor, but without CA it is not and can accumulate, so the pKa is not 6.1. - Bicarbonate secretion - Type B intercalated cells - H-ATPase polarity reversed - HCO3 Cl exchanger faces the apical rather than basolateral membrane- Titratable acidity - Weak acids are filtered at the glom and act as buffers in the urine. - HPO4 has PKA of 6.8 making it ideal - Creatinine (pKa 4.97) and uric acid (pKa 5.75) also contribute - Under normal cinditions TA buffers 10-40 mEa of H per day - Does an example of HPO4(2-):H2PO4 (1-) which exists 4:1 at pH of 7.4 (glomerular filtrate) - So for 50 mEq of Phos 40 is HPO4 and 10 is H2PO4 - When pH drops to 6.8 then the ratio is 1:1 so for 50 - So the 50 mEq is 25 and 25, so this buffered an additional 15 mEq of H while the free H+ concentration increased from 40 to 160 nanomol/L so over 99.99% of secreted H was buffered - When pH drops to 4.8 ratio is 1:100 so almost all 50 mEq of phos is H2PO4 and 39.5 mEq of H are buffered. - Acid loading decreases phosphate reabsorption so more is there to act as TA. - Decreases activity of Na-phosphate cotransporter - DKA provides a novel weak acid/buffer beta-hydroxybutyrate (pKa 4.8) which buffers significant amount of acid (50 mEq/d).- Ammonium Excretion - Ability to excrete H+ as ammonium ions adds an important amount of flexibility to renal acid-base regulation - NH3 and NH4 production and excretion can be varied according to physiologic need. - Starts with NH3 production in tubular cells - NH3, since it is neutral then diffuses into the tubule where it is acidified by the low pH to NH4+ - NH4+ is ionized and cannot cross back into the tubule cells(it is trapped in the tubular fluid) - This is important for it acting as an important buffer eve though the pKa is 9.0 - At pH of 6.0 the ratio of NH3 to NH4 is 1:1000 - As the neutral NH3 is converted to NH4 more NH3 from theintracellular compartment flows into the tubular fluid replacing the lost NH3. Rinse wash repeat. - This is an over simplification and that there are threemajor steps - NH4 is produced in early proximal tubular cells - Luminal NH4 is partially reabsorbed in the TAL and theNH3 is then recycled within the renal medulla - The medullary interstitial NH3 reaches highconcentrations that allow NH3 to diffuse into the tubular lumen in the medullary collecting tubule where it is trapped as NH4 by secreted H+ - NH4 production from Glutamine which converts to NH4 and glutamate - Glutamate is converted to alpha-ketoglutarate - Alpha ketoglutarate is converted to 2 HCO3 ions - HCO3 sent to systemic circulation by Na-3 HCO3 transporter - NH4 then secreted via Na-H exchanger into the lumen - NH4 is then reabsorbed by NaK2Cl transporter in TAL - NH4 substitutes for K - Once reabsorbed the higher intracellular pH causes NH4 to convert to NH3 and the H that is removed is secreted through Na-H exchanger to scavenge the last of the filtered bicarb. - NH3 diffuses out of the tubular cells into the interstitium - NH4 reabsorption in the TAL is suppressed by hyperkalemia and stimulated by chronic metabolic acidosis - NH4 recycling promotes acid clearance - The collecting tubule has a very low NH3 concentration - This promotes diffusion of NH3 into the collecting duct - NH3 that goes there is rapidly converted to NH4 allowing more NH3 to diffuse in. - Response to changes in pH - Increased ammonium excretion with two processes - Increased proximal NH4 production - This is delayed 24 hours to 2-3 days depending on which enzyme you look at - Decreased urine pH increases diffusion of ammonia into the MCD - Occurs with in hours of an acid load - Peak ammonium excretion takes 5-6 days! (Fig 11-10) - Glutamine is picked up from tubular fluid but with acidosis get Na dependent peritublar capillary glutamine scavenging too - Glutamine metabolism is pH dependent with increase with academia and decrease with alkalemia - NH4 excretion can go from 30-40 mEq/day to > 300 with severe metabolic acidosis (38 NaBicarb tabs) - Says each NH4 produces equimolar generation of HCO3 but I thought it was two bicarb for every alpha ketoglutarate?- The importance of urine pH - Though the total amount of hydrogren cleared by urine pH is insignificant, an acidic urine pH is essential for driving the reactions of TA and NH4 forward.- Regulation of renal hydrogen excretion - Net acid excretion vary inverse with extracellular pH - Academia triggers proximal and distal acidification - Proximally this: - Increased Na-H exchange - Increased luminal H-ATPase activity - Increased Na:3HCO3 cotransporter on the basolateral membrane - Increased NH4 production from glutamine - In the collecting tubules - Increased H-ATPase - Reduction of tubular pH promotes diffusion of NH3 which gets converted to NH4…ION TRAPPING - Extracellular pH affects net acid excretion through its affect on intracellular pH - This happens directly with respiratory disorders due to movement of CO2 through the lipid bilayer - In metabolic disorders a low extracellular bicarb with cause bicarb to diffuse out of the cell passively, this lowers intracellular pH - If you manipulate both low pCO2 and low Bicarb to keep pH stable there will be no change in the intracellular pH and there is no change in renal handling of acid. It is intracellular pH dependent - Metabolic acidosis - Ramps up net acid secretion - Starts within 24 hours and peaks after 5-6 days - Increase net secretion comes from NH4 - Phosphate is generally limited by diet - in DKA titratable acid can be ramped up - Metabolic alkalosis - Alkaline extracellular pH - Increased bicarb excretion - Decrease reabsorption - HCO3 secretion (pendrin) in cortical collecting tubule - Occurs in cortical intercalated cells able to insert H-ATPase in basolateral cells (rather than luminal membrane) - Normal subjects are able to secrete 1000 mmol/day of bicarb - Maintenance of metabolic alkalosis requires a defect which forces the renal resorption of bicarb - This can be chloride/volume deficiency - Hypokalemia - Hyperaldosteronism - Respiratory acidosis and alkalosis - PCO2 via its effect on intracellular pH is an important determinant of renal acid handling - Ratios he uses: - 3.5 per 10 for respiratory acidosis - 5 per 10 for respiratory alkalosis - Interesting paragraph contrasting the response to chronic metabolic acidosis vs chronic respiratory acidosis - Less urinary ammonium in respiratory acidosis - Major differences in proximal tubule cell pH - In metabolic acidosis there is decreased bicarb load so less to be reabsorbed proximally - In respiratory acidosis the increased serum bicarb increases the amount of bicarb that must be reabsorbed proximally - The increased activity of Na-H antiporter returns tubular cell pH to normal and prevents it from creating increased urinary ammonium - Mentions that weirdly more mRNA for H-Na antiporter in metabolic acidosis than in respiratory acidosis - Net hydrogen excretion varies with effective circulating volume - Starts with bicarb infusions - Normally Tm at 26 - But if you volume deplete the patient with diuretics first this increases to 35+ - Four factors explain this increased Tm for bicarb with volume deficiency - Reduced GFR - Activation of RAAS - Ang2 stim H-Na antiporter proximally - Ang2 also stimulates Na-3HCO3 cotransporter on basolateral membrane - Aldosterone stimulates H-ATPase in distal nephron - ALdo stimulates Cl HCO3 exchanger on basolateral membrane - Aldo stimulates eNaC producing tubular lumen negative charge to allow H secretion to occur and prevents back diffusion - Hypochloremia - Increases H secretion by both Na-dependent and Na-independent methods - If Na is 140 and Cl is 115, only 115 of Na can be reabsorbed as NaCl, the remainder must be reabsorbed with HCO3 or associated with secretion of K or H to maintained electro neutrality - This is enhanced with hypochloridemia - Concurrent hypokalemia - Changes in K lead to trans cellular shifts that affect inctracellular pH - Hypokalemia causes K out, H in and in the tubular cell the cell acts if there is systemic acidosis and increases H secretion (and bicarbonate resorption) - PTH - Decreases proximal HCO3 resorption - Primary HyperCard as cause of type 2 RTA - Does acidosis stim PTH or does PTH stim net acid excretion

If you do a Google image search for Maria Mitchell, one of the top results will be a portrait of her elegantly dressed, seated, and looking though a telescope. This image romanticizes and perpetuates the story of Maria as a lone astronomer on a distant island discovering a comet and being the first woman awarded the gold medal from the King of Denmark. But like most success stories, it's not nearly that simple.Credits:This has been a production of the Nantucket Atheneum. Written, edited and narrated by Janet Forest Special thanks to the Atheneum's Reference Library Associate Jim Borzilleri and Historian and Deputy Director of the Maria Mitchell Association Jascin Leonardo Finger for their research and insights.Voice of Alexander Dallas Bache performed by Andrew Cromartie.Resources and additional information:•Click HERE for an image of the Bonds' American Method Device. Source: Collection of Historical Scientific Instruments, Department of the History of Science, Harvard University• More on the fraying of the relationship between the Bonds and the US Coast Survey (USCS)The 1851 Exhibition marked a turning point in Bache's relationship with the Bonds. Bache had previously considered the Bonds as overly compliant towards Airy and the Liverpool Observatory during their longitude work for the USCS, and now he quietly began looking for an alternative to the Harvard College Observatory (HCO).  The situation worsened when George Bond got into a scientific fight with several of Bache's allies around 1854; a few had Harvard and USCS/Nautical Almanac connections, and almost all were part of the "Lazzaroni" group headed by Prof. Agassiz at Harvard.  When the Lazzaroni pushed the creation of the National Academy of Sciences in early 1863, George Bond was conspicuously excluded.That said, most of Bache's attempts to cut out the Bonds ended in failure. HCO and Bond & Sons kept innovating and improving their products, and the HCO continued to be a significant "default" latitude point for North America.   It was only ten years later, with the start of the Civil War, that Bache finally succeeded. Matthew Maury, the head of the US Naval Observatory joined the Confederacy, and a Bache ally took over the USNO. The Nautical Almanac moved to Washington, and coastal survey work ended, leaving the HCO isolated and underfunded. Most of the HCO's assistants enlisted for the Union, and George Bond, now widowed and working almost alone, gave up coffee and newspapers to help cut costs. The HCO was still doing important work and was critical to the success of Bond & Sons, but the center of gravity had shifted to the latter, now run by Richard Bond and John Clinch. The Nantucket Atheneum is located at 1 India Street in Nantucket, MA.You can visit us online at www.nantucketatheneum.org 

PDFchem: A new fast method to determine ISM properties and infer environmental parameters using probability distributions by Thomas G. Bisbas et al. on Friday 25 November Determining the atomic and molecular content of the interstellar medium (ISM) as a function of environmental parameters is of fundamental importance to understand the star-formation process across the epochs. Although there exist various three-dimensional hydro-chemical codes modelling the ISM at different scales and redshifts, they are computationally expensive and inefficient for studies over a large parameter space. Building on our earlier approach, we present PDFchem, a novel algorithm that models the cold ISM at moderate and large scales using functions connecting the quantities of the local ($A_{rm V,eff}$) and the observed ($A_{rm V,obs}$) visual extinctions, and the local number density, $n_{rm H}$, with probability density functions (PDF) of $A_{rm V,obs}$ on cloud scales typically tens-to-hundreds of pc as an input. For any given $A_{rm V,obs}$-PDF, sampled with thousands of clouds, the algorithm instantly computes the average abundances of the most important species (HI, H$_2$, CII, CI, CO, OH, OH$^+$, H$_2$O$^+$, CH, HCO$^+$) and performs radiative transfer calculations to estimate the average emission of the most commonly observed lines ([CII]~$158mu$m, both [CI] fine-structure lines and the first five rotational transitions of $^{12}$CO). We examine two $A_{rm V,obs}$-PDFs corresponding to a non star-forming and a star-forming ISM region, under a variety of environmental parameters combinations. These cover FUV intensities in the range of $chi/chi_0=10^{-1}-10^3$, cosmic-ray ionization rates in the range of $zeta_{rm CR}=10^{-17}-10^{-13},{rm s}^{-1}$ and metallicities in the range of $Z=0.1-2,{rm Z}_{odot}$. PDFchem is fast, easy to use, reproduces the PDR quantities of the time-consuming hydrodynamical models and can be used directly with observed data to understand the evolution of the cold ISM chemistry. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.12974v1

PDFchem: A new fast method to determine ISM properties and infer environmental parameters using probability distributions by Thomas G. Bisbas et al. on Thursday 24 November Determining the atomic and molecular content of the interstellar medium (ISM) as a function of environmental parameters is of fundamental importance to understand the star-formation process across the epochs. Although there exist various three-dimensional hydro-chemical codes modelling the ISM at different scales and redshifts, they are computationally expensive and inefficient for studies over a large parameter space. Building on our earlier approach, we present PDFchem, a novel algorithm that models the cold ISM at moderate and large scales using functions connecting the quantities of the local ($A_{rm V,eff}$) and the observed ($A_{rm V,obs}$) visual extinctions, and the local number density, $n_{rm H}$, with probability density functions (PDF) of $A_{rm V,obs}$ on cloud scales typically tens-to-hundreds of pc as an input. For any given $A_{rm V,obs}$-PDF, sampled with thousands of clouds, the algorithm instantly computes the average abundances of the most important species (HI, H$_2$, CII, CI, CO, OH, OH$^+$, H$_2$O$^+$, CH, HCO$^+$) and performs radiative transfer calculations to estimate the average emission of the most commonly observed lines ([CII]~$158mu$m, both [CI] fine-structure lines and the first five rotational transitions of $^{12}$CO). We examine two $A_{rm V,obs}$-PDFs corresponding to a non star-forming and a star-forming ISM region, under a variety of environmental parameters combinations. These cover FUV intensities in the range of $chi/chi_0=10^{-1}-10^3$, cosmic-ray ionization rates in the range of $zeta_{rm CR}=10^{-17}-10^{-13},{rm s}^{-1}$ and metallicities in the range of $Z=0.1-2,{rm Z}_{odot}$. PDFchem is fast, easy to use, reproduces the PDR quantities of the time-consuming hydrodynamical models and can be used directly with observed data to understand the evolution of the cold ISM chemistry. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.12974v1

Mapping NGC 7027 in New Light: CO ^+ and HCO ^+ Emission Reveal Its Photon- and X-ray-Dominated Regions by Jesse Bublitz et al. on Wednesday 23 November The young and well-studied planetary nebula NGC 7027 harbors significant molecular gas that is irradiated by luminous, point-like UV (central star) and diffuse (shocked nebular) X-ray emission. This nebula represents an excellent subject to investigate the molecular chemistry and physical conditions within photon- and X-ray-dominated regions (PDRs and XDRs). As yet, the exact formation routes of CO$^+$ and HCO$^+$ in PN environments remain uncertain. Here, we present $sim$2$"$ resolution maps of NGC 7027 in the irradiation tracers CO$^+$ and HCO$^+$, obtained with the IRAM NOEMA interferometer, along with SMA CO and HST 2.12~$mu$m H$_2$ data for context. The CO$^+$ map constitutes the first interferometric map of this molecular ion in any PN. Comparison of CO$^+$ and HCO$^+$ maps reveal strikingly different emission morphologies, as well as a systematic spatial displacement between the two molecules; the regions of brightest HCO$^+$, found along the central waist of the nebula, are radially offset by $sim$1$"$ ($sim$900 au) outside the corresponding CO$^+$ emission peaks. The CO$^+$ emission furthermore precisely traces the inner boundaries of the nebula's PDR (as delineated by near-IR H$_2$ emission), suggesting that central star UV emission drives CO$^+$ formation. The displacement of HCO$^+$ radially outward with respect to CO$^+$ is indicative that dust-penetrating soft X-rays are responsible for enhancing the HCO$^+$ abundance in the surrounding molecular envelope, forming an XDR. These interferometric CO$^+$ and HCO$^+$ observations of NGC 7027 thus clearly establish the spatial distinction between the PDR and XDR formed (respectively) by intense UV and X-ray irradiation of molecular gas. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.13680v2

Mapping NGC 7027 in New Light: CO ^+ and HCO ^+ Emission Reveal Its Photon- and X-ray-Dominated Regions by Jesse Bublitz et al. on Tuesday 22 November The young and well-studied planetary nebula NGC 7027 harbors significant molecular gas that is irradiated by luminous, point-like UV (central star) and diffuse (shocked nebular) X-ray emission. This nebula represents an excellent subject to investigate the molecular chemistry and physical conditions within photon- and X-ray-dominated regions (PDRs and XDRs). As yet, the exact formation routes of CO$^+$ and HCO$^+$ in PN environments remain uncertain. Here, we present $sim$2$"$ resolution maps of NGC 7027 in the irradiation tracers CO$^+$ and HCO$^+$, obtained with the IRAM NOEMA interferometer, along with SMA CO and HST 2.12~$mu$m H$_2$ data for context. The CO$^+$ map constitutes the first interferometric map of this molecular ion in any PN. Comparison of CO$^+$ and HCO$^+$ maps reveal strikingly different emission morphologies, as well as a systematic spatial displacement between the two molecules; the regions of brightest HCO$^+$, found along the central waist of the nebula, are radially offset by $sim$1$"$ ($sim$900 au) outside the corresponding CO$^+$ emission peaks. The CO$^+$ emission furthermore precisely traces the inner boundaries of the nebula's PDR (as delineated by near-IR H$_2$ emission), suggesting that central star UV emission drives CO$^+$ formation. The displacement of HCO$^+$ radially outward with respect to CO$^+$ is indicative that dust-penetrating soft X-rays are responsible for enhancing the HCO$^+$ abundance in the surrounding molecular envelope, forming an XDR. These interferometric CO$^+$ and HCO$^+$ observations of NGC 7027 thus clearly establish the spatial distinction between the PDR and XDR formed (respectively) by intense UV and X-ray irradiation of molecular gas. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.13680v2

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.04.515240v1?rss=1 Authors: Dwivedi, I., Zhou, D., Caldwell, A. B., Subramaniam, S., Haddad, G. G. Abstract: Opioid use disorder (OUD) among pregnant women has become an epidemic in the United States. Pharmacological interventions for OUD involve methadone, a synthetic opioid analgesic that attenuates withdrawal symptoms and behaviors linked with maternal drug addiction. However, methadone's ability to readily accumulate in neural tissue, and cause long-term neurocognitive sequelae, has led to concerns regarding its effect on prenatal brain development. We took advantage of human cortical organoid (hCO) technology to probe how this drug impacts the earliest mechanisms giving rise to the cerebral cortex. To this end, we conducted bulk mRNA sequencing of 2-month-old hCOs derived from two cell lines that were chronically treated with a clinically relevant dose of 1M methadone for 50 days. Differential expression and gene ontology analyses revealed a robust transcriptional response to methadone associated with functional components of the synapse, the underlying extracellular matrix (ECM), and cilia. Further unsupervised co-expression network and predictive protein-protein interaction analyses demonstrated that these changes occurred in concert, centered around a regulatory axis consisting of growth factors, developmental signaling pathways, and matricellular proteins. Our results demonstrate that exposure to methadone during early cortico-genesis fundamentally alters transcriptional programs associated with synapse formation, and that these changes arise by modulating extra-synaptic molecular mechanisms in the ECM and cilia. These findings provide novel insight into methadone's putative effect on cognitive and behavioral development and a basis for improving interventions for maternal opioid addiction. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

The evolution of HCO ^ + in molecular clouds using a novel chemical post-processing algorithm by M. Panessa et al. on Thursday 13 October Modeling the internal chemistry of molecular clouds is critical to accurately simulating their evolution. To reduce computational expense, 3D simulations generally restrict their chemical modeling to species with strong heating and cooling effects. We address this by post-processing tracer particles in the SILCC-Zoom molecular cloud simulations. Using a chemical network of 39 species and 299 reactions (including freeze-out of CO and H$_2$O), and a novel iterative algorithm to reconstruct a filled density grid from sparse tracer particle data, we produce time-dependent density distributions for various species. We focus upon the evolution of HCO$^+$, which is a critical formation reactant of CO but is not typically modeled on-the-fly. We analyse the evolution of the tracer particles to assess the regime in which HCO$^+$ production preferentially takes place. We find that the HCO$^+$ content of the cold molecular gas forms in situ around $n_textrm{HCO$^+$}simeq10^3$-$10^4$ cm$^{-3}$, over a time-scale of approximately 1 Myr, rather than being distributed to this density regime via turbulent mixing from deeper in the cloud. We further show that the dominant HCO$^+$ formation pathway is dependent on the visual extinction, with the reaction H$_3^+$ + CO contributing 90% of the total HCO$^+$ production flux above $A_textrm{V,3D}=3$. Using our novel grid reconstruction algorithm, we produce the very first maps of the HCO$^+$ column density, $N$(HCO$^+$), and show that it reaches values as high as $10^{15}$ cm$^{-2}$. We find that 50% of the HCO$^+$ mass is located in an $A_textrm{V}$-range of $sim$10-30, and in a density range of $10^{3.5}$-$10^{4.5}$ cm$^{-3}$. Finally, we compare our $N$(HCO$^+$) maps to recent observations of W49A and find good agreement. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2210.06251v1

The evolution of HCO ^ + in molecular clouds using a novel chemical post-processing algorithm by M. Panessa et al. on Thursday 13 October Modeling the internal chemistry of molecular clouds is critical to accurately simulating their evolution. To reduce computational expense, 3D simulations generally restrict their chemical modeling to species with strong heating and cooling effects. We address this by post-processing tracer particles in the SILCC-Zoom molecular cloud simulations. Using a chemical network of 39 species and 299 reactions (including freeze-out of CO and H$_2$O), and a novel iterative algorithm to reconstruct a filled density grid from sparse tracer particle data, we produce time-dependent density distributions for various species. We focus upon the evolution of HCO$^+$, which is a critical formation reactant of CO but is not typically modeled on-the-fly. We analyse the evolution of the tracer particles to assess the regime in which HCO$^+$ production preferentially takes place. We find that the HCO$^+$ content of the cold molecular gas forms in situ around $n_textrm{HCO$^+$}simeq10^3$-$10^4$ cm$^{-3}$, over a time-scale of approximately 1 Myr, rather than being distributed to this density regime via turbulent mixing from deeper in the cloud. We further show that the dominant HCO$^+$ formation pathway is dependent on the visual extinction, with the reaction H$_3^+$ + CO contributing 90% of the total HCO$^+$ production flux above $A_textrm{V,3D}=3$. Using our novel grid reconstruction algorithm, we produce the very first maps of the HCO$^+$ column density, $N$(HCO$^+$), and show that it reaches values as high as $10^{15}$ cm$^{-2}$. We find that 50% of the HCO$^+$ mass is located in an $A_textrm{V}$-range of $sim$10-30, and in a density range of $10^{3.5}$-$10^{4.5}$ cm$^{-3}$. Finally, we compare our $N$(HCO$^+$) maps to recent observations of W49A and find good agreement. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2210.06251v1

John Foster, 4-time CHRO & Founder of Gamut Labs, joined us on The Modern People Leader. We talked about the difference between a traditional and a human-centered organization, the 5 dimensions of a HCO, and why HCOs get more creative output from their people. Timestamps: (11:23) The difference between a traditional organization and a Human-Centered Organization (16:28) Why a little bit of hierarchical thinking can ruin a people process (25:25) The definition of a Human-Centered Organization (28:25) The 5 dimensions of a Human-Centered Organization (41:07) The best way to get creative output is by being human-centered (44:01) Why the traditional organization is a crack in the sidewalk of humanity (46:33) How John defines a Modern People Leader Subscribe to the MPL Weekly Digest: https://forms.gle/qdt6YaWULfoEHb6n8 Follow us on LinkedIn: https://www.linkedin.com/company/the-modern-people-leader

Understanding if molecular ratios can be used as diagnostics of AGN and starburst activity: The case of NGC 1068 by J. Butterworth et al. on Thursday 22 September Molecular line ratios, such as HCN(1-0)/HCO$^+$(1-0) and HCN(4-3)/CS(7-6), are routinely used to identify active galactic nuclei (AGN) activity in galaxies. Such ratios are, however, hard to interpret as they are highly dependent on the physics and energetics of the gas, and hence can seldom be used as a unique, unambiguous diagnostic. We used the composite galaxy NGC 1068 as a `laboratory' to investigate whether molecular line ratios between HCN, HCO$^+$, and CS are useful tracers of AGN-dominated gas and determine the origin of the differences in such ratios across different types of gas. Such a determination will enable a more rigorous use of such ratios. First, we empirically examined the aforementioned ratios at different angular resolutions to quantify correlations. We then used local thermodynamic equilibrium (LTE) and non-LTE analyses coupled with Markov chain Monte Carlo (MCMC) sampling in order to determine the origin of the underlying differences in ratios. We propose that at high spatial resolution (< 50 pc) the HCN(4-3)/CS(2-1) is a reliable tracer of AGN activity. We also find that the variations in ratios are not a consequence of different densities or temperature but of different fractional abundances, yielding to the important result that it is essential to consider the chemical processes at play when drawing conclusions from radiative transfer calculations. From analyses at varying spatial scales, we find that previously proposed molecular line ratios, as well as a new one, have varying levels of consistency. We also determine from an investigation of radiative transfer modelling of our data that it is essential to consider the chemistry of the species when reaching conclusions from radiative transfer calculations. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.05928v2

Understanding if molecular ratios can be used as diagnostics of AGN and starburst activity: The case of NGC 1068 by J. Butterworth et al. on Thursday 22 September Molecular line ratios, such as HCN(1-0)/HCO$^+$(1-0) and HCN(4-3)/CS(7-6), are routinely used to identify active galactic nuclei (AGN) activity in galaxies. Such ratios are, however, hard to interpret as they are highly dependent on the physics and energetics of the gas, and hence can seldom be used as a unique, unambiguous diagnostic. We used the composite galaxy NGC 1068 as a `laboratory' to investigate whether molecular line ratios between HCN, HCO$^+$, and CS are useful tracers of AGN-dominated gas and determine the origin of the differences in such ratios across different types of gas. Such a determination will enable a more rigorous use of such ratios. First, we empirically examined the aforementioned ratios at different angular resolutions to quantify correlations. We then used local thermodynamic equilibrium (LTE) and non-LTE analyses coupled with Markov chain Monte Carlo (MCMC) sampling in order to determine the origin of the underlying differences in ratios. We propose that at high spatial resolution (< 50 pc) the HCN(4-3)/CS(2-1) is a reliable tracer of AGN activity. We also find that the variations in ratios are not a consequence of different densities or temperature but of different fractional abundances, yielding to the important result that it is essential to consider the chemical processes at play when drawing conclusions from radiative transfer calculations. From analyses at varying spatial scales, we find that previously proposed molecular line ratios, as well as a new one, have varying levels of consistency. We also determine from an investigation of radiative transfer modelling of our data that it is essential to consider the chemistry of the species when reaching conclusions from radiative transfer calculations. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.05928v2

Can Molecular Ratios be used as Diagnostics of AGN and Starburst activity? The Case of NGC 1068 by J. Butterworth et al. on Tuesday 13 September Molecular line ratios, such as HCN(1-0)/HCO$^+$(1-0) and HCN(4-3)/CS(7-6) are routinely used to identify AGN activity in galaxies. Such ratios are however hard to interpret as they are highly dependent on the physics and energetics of the gas and hence can seldom be used a as a unique unambiguous diagnostic. We use the composite galaxy NGC 1068 as a "laboratory", to investigate whether molecular line ratios between HCN, HCO$^+$ and CS are useful tracers of AGN-dominated gas and determine the origin of the differences in such ratios across different types of gas. Such determination will allow a more rigorous use of such ratios. We first empirically examine the aforementioned ratios at different angular resolutions to quantify correlations. We then use LTE and non-LTE analyses coupled with Markov Chain Monte Carlo (MCMC) sampling in order to determine the origin of the underlying differences in ratios. We propose that at high spatial resolution (< 50 pc) the HCN(4-3)/CS(2-1) is a reliable tracer of AGN activity. Finally we find that the variations in ratios are not a consequence of different densities or temperature but of different fractional abundances yielding to the important result that it is essential to consider the what chemical processes are at play when drawing conclusions from radiative transfer calculations. Upon analysis at varying spatial scales previous proposed as well as a new molecular line ratio have been shown to have varying levels of consistency. We have also determined from investigation of radiative transfer modelling of our data that it is essential to consider the chemistry of the species when reaching conclusions from radiative transfer calculations. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.05928v1

Tracing the contraction of the pre-stellar core L1544 with HC ^ 17 O ^+ J = 1-0 emission by J. Ferrer Asensio et al. on Tuesday 06 September Spectral line profiles of several molecules observed towards the pre-stellar core L1544 appear double-peaked. For abundant molecular species this line morphology has been linked to self-absorption. However, the physical process behind the double-peaked morphology for less abundant species is still under debate. In order to understand the cause behind the double-peaked spectra of optically thin transitions and their link to the physical structure of pre-stellar cores, we present high-sensitivity and high-spectral resolution HC$^{17}$O$^+$ $J =$1-0 observations towards the dust peak in L1544. We observed the HC$^{17}$O$^+$ (1-0) spectrum with the Institut de Radioastronomie Millim'etrique (IRAM) 30m telescope. By using new state-of-the-art collisional rate coefficients, a physical model for the core and the fractional abundance profile of HC$^{17}$O$^+$, the hyperfine structure of this molecular ion is modelled for the first time with the radiative transfer code LOC applied to the predicted chemical structure of a contracting pre-stellar core. We applied the same analysis to the chemically related C$^{17}$O molecule. The observed HC$^{17}$O$^+$(1-0) and C$^{17}$O(1-0) lines have been successfully reproduced with a non-local thermal equilibrium (LTE) radiative transfer model applied to chemical model predictions for a contracting pre-stellar core. An upscaled velocity profile (by 30%) is needed to reproduce the HC$^{17}$O$^+$(1-0) observations. The double peaks observed in the HC$^{17}$O$^+$(1-0) hyperfine components are due to the contraction motions at densities close to the critical density of the transition ($sim$10$^{5}$ cm$^{-3}$) and to the fact that the HCO$^{+}$ fractional abundance decreases toward the centre. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2209.02489v1

*** WELCOME TO OUR 100th Episode of Homeopathy Hangout Podcast! Please support our sponsors on their social links below *** Today's super special guest is Miranda Castro - a British homeopath who has been practicing classical homeopathy since 1982. She is author of the best-selling "The Complete Homeopathy Handbook" (over 500,000 copies sold), the much-loved Homeopathy for Mother and Baby and A Homeopathic Guide to Stress.  Miranda believes passionately that homeopathy is 'medicine for the people' and her writing reflects that: her numerous articles and her books have made classical homeopathy spectacularly accessible to the lay person (home prescriber?)—giving people the tools to accurately select healing remedies for themselves and their families.  She has the ability to communicate homeopathic topics in an engaging and accessible manner. Her infectious enthusiasm comes through in all her articles, lectures, webinars … and podcasts. She moved to the US in 1994 and currently lives in Florida. She can be contacted at mirandacastro.com A big thank you to the sponsors for our 100th episode Celebration Party in Perth today. Please follow/like them and support them if you can. We are so grateful to them for making today's party super special! - A Woman's touch Photography: instagram.com/awomanstouchphotography/or facebook.com/awomanstouchphotography - Bite into Bliss: facebook.com/biteintothebliss or instagram.com/bitein2bliss/ - Classes Cakes and More: facebook.com/classescakesandmore or instagram.com/classescakesandmore/ - Chaga Health: instagram.com/chagahealth_au or Facebook.com/ChagaHealthAustralia - Clean Minds - no socials, but Anna can be reached on 042 69 72 389 and produces healing stories, cleaning services and placenta prints - Dinner Twist: facebook.com/dinnertwist or instagram.com/dinnertwist - Cakealicious: facebook.com/CakealiciousWA or instagram.com/cakealicious58 - De Light Me Cocktail Cookies: instagram.com/de.light_me - Em Energetic: instagram.com/em.energetic/ or facebook.com/em.energetic - IconFit: facebook.com/iconfitaustralia or instagram.com/iconfitaustralia/ - Just Add Health: facebook.com/JustAddHealthStoneville or instagram.com/justaddhealth - Heather & Co Floral Artisans: instagram.com/handco.perth/ or facebook.com/HCo-104044151916915 - Happy Patch Herbal and Healing: facebook.com/happypatchherbals or instagram.com/happy_patch_herbals/ - Little Miss Organic: instagram.com/little_miss_organic or facebook.com/LittleMissOrganic - Little Miss Enjo: instagram.com/little_miss_enjo or facebook.com/LittleMissENJO - Metta Chiropractic: instagram.com/mettachiropractichealth/ or facebook.com/MettaChiro/ - Macrocos Mish Intuitive Healer: facebook.com/MacrocosMish or instagram.com/macrocosmish - Mother Me Postpartum Meal delivery: instagram.com/_motherme/ or facebook.com/mothermeperth/ - Nourished by Broth: facebook.com/Nourishedbybroth/ - Powder Candle: Instagram.com/powdercandleaustralia or facebook.com/PowderCandleAustralia - The Milk Bar Candle Company:facebook.com/The-Milk-Bar-Candle-Co-102558728992862 or instagram.com/themilkbarcandleco - The Conscious Caregiver: instagram.com/consciousmothersgroup

“The hidden harmony is better than the obvious.” Heraclitus   Earlier this year RESNET was approved as a Home Certification Organization (HCO) for the California ENERGY STAR Homes Program by the Environmental Protection Agency (EPA).   What are the roots of this partnership with CalCERTS?   Who will benefit from this harmony?   What benefits will evolve for the parties involved? We are joined today by David Choo, Director, Compliance at CalCERTS and Laurel Elam, Director of Quality Assurance and Standards at RESNET to get the details behind the fact that RESNET HERS index ratings are soon to be recognized in California.   We will learn why RESNET chose to do a pilot in California, what this means to CalCERTS and how it works.   Amongst several other topics of discussion, David will give us insights into the demand for this program and Laurel will discuss what an HCO is as well as how it relates to RESNET.   This episode will be of special interest to national production builders that build in California.   You can learn a more about the background of this topic in RESTalk Episode 79 entitled California Dreamin'   For more information about the California ENERGY STAR program requirements please email support@calcerts.com.   Also see this article on the RESNET site: https://www.resnet.us/articles/resnet-hers-index-ratings-soon-to-be-recognized-in-california/     RESTalk: To the RESNET community, we hear you and want to engage. Learn more at www.RESNET.us Or for more info on this topic contact RESNET at INFO@RESNET.US

MRG address issues involving access to care, chronic disease management, patient adherence & compliance to doctors orders, and frees up clinicians time through leveraging technology so they can better monitor and manage larger amounts of patients more effectively and efficiently.Smart Care 360 is a complete and comprehensive care management platform that allows physicians and caregivers to manage the full continuum of care. By continuous monitoring of a patient's vitals, nutrition, medication, and daily living activities clinicians see a holistic picture of the patient. Using their AI and threshold capabilities they provide a more proactive approach for care decisions and can alert caregivers to any change in condition. The platform is a complete solution that includes auto messaging, reminders to patients, 2 way communication, video conferencing with vitals displayed in real time -- all recorded and integrated into any EMR. With MRG Care 360, there is a single view of everything on a patient, with alerts, thresholds, and continuous access for patients and physicians to each other. The MRG real-time RPM Management Solution increases staffing efficiency, reduces doctor and nurse burnout and provides more patient info with instant access to the patients and staff. The platform was developed after 2+ years of R&D, and being involved in 4 very in depth Texas based studies on major clinical / operational needs in specific specialties & chronic diseases throughout rural, urban, and metro geographic regions. The data was compiled, and prioritization of our capabilities was built on a code-less drag and drop UX/UI with decision tree logic that allows simple patient planning, customization and flexibility based on the HCO's needs which also promotes efficient and scalable deployment solutions for large variable patient groups based on the specific specialty, disease, or vertical market needs.

This week we sit down with Gravel Locos founder Fabian Serralta to unpack the road to developing a stand out event.  New for this year, Gravel Locos will be adding an event in Peublo, CO in addition to the original event in Hico, TX. Episode Sponsor: Hammerhead.  Use coupon code 'TheGravelRide' for a free custom color kit and premium water bottle. Gravel Locos Website Support the Podcast Join The Ridership  Automated Transcription, please excuse the typos: Gravel Locos [00:00:00] Craig Dalton: Hello, and welcome to the gravel ride podcast, where we go deep on the sport of gravel cycling through in-depth interviews with product designers, event organizers and athletes. Who are pioneering the sport I'm your host, Craig Dalton, a lifelong cyclist who discovered gravel cycling back in 2016 and made all the mistakes you don't need to make. I approach each episode as a beginner down, unlock all the knowledge you need to become a great gravel cyclist.   [00:00:26] Craig Dalton: Have you ever thought about organizing a gravel event? I certainly have this week's guest Fabion. Serralta took that passion and idea to create an event. And created gravel Locos. The original event in Heico Texas is joined this year by a new event in Pueblo, Colorado. I sat down with Fabi and to learn about his inspiration for gravel Locos, the charitable component of what he does and the general theme of all gravel Locos events. Before we jump into the conversation. I need to thank this week. Sponsor hammerhead. The hammerhead kuru to you as raised the bar for cycling computers. You can get advanced GPS, navigation, and intuitive software right on your handlebars. In a way you never thought possible. I spent this past weekend down in Tucson, Arizona. After speaking with John from the mountain lemon, gravel grinder, a few years back, I've been itching to get out on the course. So I simply downloaded the GPS file from their website, uploaded it to my kuru to and felt super confident going out there in the back country. I've got a ton of stories about some of the snafoos. I had mechanically speaking while I was out there, but from a navigational perspective, it was spot on what I really appreciate about the crew too. And I've talked about the responsive touchscreen display before. Is that in the navigation? Once you've loaded a route in there, you get a little icon, no matter what screen you're on. saying when the next turn is coming up. It's those little touches and details that I think really setting the hammerhead crew to a part beyond that, I really got to dive into the climber feature, which is something unique and special about hammerhead. The climber feature allows you to visualize and prepare for upcoming gradient changes in real time. So, what does that mean? Basically it translates to a nice graph. On the climber screen on your crew to computer that shows you in color-coded fashion. The length to the top of the climb, both in miles, as well as elevation, and then gradient by gradient profile looks that map exactly to what you're experiencing when you're out on an adventure loop that you've never been on before. It was super useful to see, okay, this is going to be a punchy. Mile mile and a half climb. Versus at the end i discovered as it turned a corner that i was in front of a six mile climb but fortunately the gradient was pretty chill. This all translates to knowledge is power. And with the hammerhead crew too, you can get all the information available out of your GPX files. You can customize it to the nth degree. I still have a ways to go and customizing mine, but you can see the power of organizing your data right there on your computer screen. For a limited time, our listeners can get a free custom color kit and exclusive premium water bottle with the purchase of a hammerhead crew to visit hammerhead.io right now and use the promo code, the gravel ride. At checkout to get yours today, that's a free custom color kit. And a premium water bottle with the purchase of a career to. Go to hammerhead.io at all three items to your cart and use the promo code. The gravel ride. With that said let's jump right into my interview with Fabienne from gravel Locos. [00:03:40] CraigDalton.: Fabian , welcome to the show. [00:03:41] FabianSerralta: Hey, Craig. Thank you. [00:03:43] CraigDalton.: It's good to talk to you again. [00:03:44] FabianSerralta: Same here. Thank you. [00:03:46] CraigDalton.: I'm excited to learn all the things about gravel locus and, but would love to learn a little bit about your background first, just kinda what drew you into the sport originally, and then what led you to take on the huge challenge of creating an event? [00:04:01] FabianSerralta: Well, I, I would definitely say I was probably what led me to the sport was purely accidental. I I purchased a, a ranch in Oklahoma in 2012, and I remember the first time driving out there to see this ranch with the R I was following a realtor and wearing this perfectly smooth country road. And I was just thinking, this is great. I'm gonna have my road bike out here. And and as soon as we get to the one road leads to the ranch, it was this. Awful road with rocks and gravel and dirt. And I was like, oh my gosh, this is terrible. I'm not gonna be able to ride my road, bike out here. And then we're on this road. And this lady is just like flying on this road and just dusting, dirt everywhere. And then we're just flying on their road and, and it was like a 15 mile drive from on this. Awful gravel dirt road to the driveway of this ranch that I had just purchased and or I was about to purchase. And then the driveway from this road to the ranch was another mile and a half. And when we get to the ranch property in the house, I said to her. This is terrible. I'm not gonna be able to ride my bike when I'm out here. She's like, well, what do you mean? I said, I'll get a flat tire by the time I get to, to the to the main road. And she's like, well, you can just drive your bike in your car and, and go out there and just park out there. I'm like, where am I gonna park? That's like somebody else's property out there. So this is 2012. And I, so I buy this property and probably from 20 12, 20 13 until about 2015. I didn't get to ride my road bike every time that I went out there. [00:05:41] CraigDalton.: yeah. I was gonna say there wasn't a lot of options [00:05:43] FabianSerralta: Yeah. [00:05:44] CraigDalton.: of gravel bikes at that point in time. [00:05:45] FabianSerralta: Yeah. But then in 2015, I'm visiting my son in Denver. He was gonna school at the university of Denver and I just happened to go to a, a bike store. It was a, I believe it was a specialized bike store and I walk in there and there's. White and red bike Witham, and it looked like a road bike and it had these Nobby tires on it and zip wheels. And I'm like, what in the world is this thing? And the sales guy comes up to me and I'm like, what is this thing? You know? And he's looking at me like, he's like, where are you from? I'm like, well, I'm from Cuba. And he's probably thinking this guy just fell off of open a Palm or something. Cause likes a, and I'm like, what's a, you, I had idea what a. He's like, you don't know what a cross bike is. I was like, I don't know, but I want that bike. He's like, well, you gonna Doy lacrosse. I'm like, no, I bought this ranch and it has these horrible roads that are rocks. And he's like, oh, so you want a gravel bike? I'm like a gravel bike. I said, no, I want that bike. And I need you to ship it to Texas. And the guy is just like, looking me, like I'm nuts. But it was like, It looked like a road bike and it had, you know, what looked like to be mountain bike tires. And I'm like, this is it. So I purchased a bike and it happened to be on my size, a 54 centimeter. They ship it to to Texas to my local bike shop and gets over there. They had 700 by 33 C tires. I start writing it in Oklahoma and I thought was the greatest thing ever. And I was running 90 PSI on the tires, which I thought at the time was perfect. As you know, on my tubular road tires, I was running 120 PSI. And so I thought this thing was just as smooth as can be on these gravel roads. And I signed up for my first gravel event, October 1st, 2016. And I thought, Hey, you know, it was 15 miles, no big deal. I showed up at the time, I think I had 95 P assigned a rear and 90 in the front. And I thought it was gonna be very much like a road rally where you have, you know, rest stops every so many miles. And so I think I had two water bottles and, or maybe a one snack or whatever, and man, it was a nightmare. [00:07:55] CraigDalton.: Was was that in Oklahoma, the event that you signed [00:07:58] FabianSerralta: it was Texas. It was a really hilly area monster, Texas. And you know, I paid my registration fee, which I think it was, I don't know, 85, $90. And I was expecting it to be just like many of the road rallies I had done. And. You know, grass stops with bananas and oranges and cookies and pickle, juices, and Gatorade. And there was nothing. all they had was a water table. with warm water, no food. And by the end of the 50 miles, I thought I was gonna die. And with that tire pressure on those 33 sea tires. I was so beat up. I swear, I'll never do this again. I remember getting to the finish and calling some buddies. I'm like, I am never doing this again. This is horrible. Why would anybody ever do this? You know, [00:08:43] CraigDalton.: Yeah. It's, it's funny. What a difference the evolution of the equipment has made in the enjoyment of the sport. I'm with you. I, I got my first gravel bike a little bit later than the, I think back in 2016, but it was a. 2014 model year bike, 32 C tires. Fortunately it, it did have tubeless on it, but it still, like, I just felt like it wasn't that much better than Ayro bike. I had five or six years earlier, which I had pretty much quit riding because I would either flat or get the crap beat outta me every time I rode it. [00:09:15] FabianSerralta: Yeah, this, this, I mean, it was, it was ay, it was a cycle cross bike. It was specialized crux. I mean, it had great, I mean, it had zip three or threes. I was running tubes. Which was, you know, a big difference from running tubers on my other bikes road bikes. And they had ceramic red, I couldn't complain it was a great bike, but I was just running to run tire pressure, the wrong tire size. And I really didn't have any, any knowledge of, of gravel, but, you know, I did see other people running big, your tires and all that. And it, it was just this learning curve. But that first experience was horrible. And I really, I swear I never did this again. And it took a while and then I, I started learning from others like, Hey, yeah, you can't be out there running tire pressure like that. And you can't, you, you gotta run bigger tires than that. And you gotta run, you know, tube, you know, tubeless. And and you know, I, it probably took me a few, a couple of months before I even wrote again. But as I, I started getting more and more advice from others that were doing it. But it was so early on. But it was that experience that really led me to wanna have eventually at one day have a gravel event. That was a lot like a road rally, but that it was also, you know, it, it had the, it was at the time it was, you know, the dirty cancer event, the DK 200. So I wanted to have a, a DK 200 event with the pro component, but yet. Beginner friendly. So have all the support that you would need for beginners like myself or, or people just really interested in gravel. So having all the bunch of rest stops and having portable bathrooms at rest stops for the women and having sag vehicles all over the place, but yet having a ton of pros. So having, you know, an event that was a DK 200 packed with pros, but yet packed with. All the support and all the things that you are accustomed to experiencing and having at a, you know, family road, rally type event. [00:11:15] CraigDalton.: Interesting. So it sounds like, I mean, if we fast forward a few years from that original event, it sounds like you competed in a handful of events every year to try to, you know, obviously continue your enjoyment of the sport. The, the equipment had continued to evolve and, and you'd had a number of experiences at other events where you're like, I like part of what this event has done, but part of what that event has done. And you thought, well, like what if I did this on my own? [00:11:41] FabianSerralta: Yeah, look so it, the events continue. I con I would go to every gravel event that I could go to, but it just, it was, you know, you pay your 75, 85, a hundred something dollars, but. You're lucky you got a water table and it was, everything was always self support, self support, self supported, and you're paying all this money, but you're not really getting a whole lot in return. And to me, it really excluded a lot of beginners. It excluded people that didn't have a lot of bike skills or, or bike mechanic knowledge because, you know, I always say what, what makes riding gravel so great is that you're out in the middle of nowhere. , but what that's also, what makes it kind of dangerous and also kind of keeps a lot of people out of it because you know, you're out in the middle of nowhere. You don't have cell reception, you don't have convenience stores. Oftentimes you don't have many houses or you don't see a vehicle or cars don't even go come by half the time. So yeah, you're out there in the middle of nowhere, but then you don't help of neighbors and you don't have convenience stores and you don't have others to reach out to in case of an emergency or a mechanical. And I feel that that deters a lot of people from venturing and, and experiencing gravel. And as a result, you know, a lot of people miss out on experiencing gravel. So how do you bring in all those people? And for me, Is the way to do that is by bringing in all the support, the sag vehicles, aid stations, every so many miles. So for example, in gravel Locos, you have, you know, six aid stations you have 20 something sag vehicles for the women. We have portable bathrooms at all the aid stations where there is in HaCo or in Pueblo, Colorado . And that's how we take out that, that fear of, you know, Being out there in the middle of nowhere. So, but yet we still have, you know, 20 something, 30 pros out there participating just like you, we did, you know, at events like dirty cans, 200 or, you know, what was land run, you know? And I referred to them by those names because that's what I was modeling. Then. [00:13:42] CraigDalton.: I'm glad we unearthed that because I think it's important to kind of think about people's orientations as event organizers in terms of how they're gonna set up the overall experience. You know, it's one thing when you've got a, a top level pro who's decided they want to get into the event business. And oftentimes they do design events that are driven from the front. They're really a professional experience that does trickle down to the rest of us. But it's you know, it's pretty refreshing to hear you talk about. Wanting the last person in the event to have the best time possible as well as, as the first. So stepping back for a second, you, you live in Arlington, Texas, you've got property in Oklahoma. You'd experience the gravel community for a number of years. You decided, Hey, there's something missing. I'd love to highlight my perspective of a gravel event. How did you decide on, on, on Texas for the original event and what was that process like? [00:14:36] FabianSerralta: Well, I, you said it in, cause that's where I predominantly ride, you know, so I've a lot gravel. I've done gravel in California. I've done gravel in Vermont, in, in Montana. I've done gravel in Scotland. I, you know, I've, I've done gravel in other parts and, but Texas, where, where I mostly write gravel and I've done gravel all over Texas and HaCo. I remember writing and close to close to HaCo. And one of the folks that I was riding with says, man, if you like this area, You've gotta check out. Heico so I said Heico where's that? So I found it went out there and, [00:15:13] CraigDalton.: And where, where is it relative to, to the Dallas area [00:15:17] FabianSerralta: for me, it's about an hour and 20 minute drive [00:15:20] CraigDalton.: and is that to, to the east or which direction [00:15:23] FabianSerralta: I'm the guy that gets lost with Garmen. So so I'm gonna take a chance here. Say I think it's so out. [00:15:30] CraigDalton.: Okay. [00:15:31] FabianSerralta: So, yeah, I'm horrible. I'm like directionally challenged. So I get asked this all the time. Like the other day I was in Pueblo for meetings and I was meeting with the the the PBR, the folks for the professional bull riding association, which one of our sponsors in there were asking me. So which direction I, I have no idea. Here's the route, you figure it out. And really, I do get lost even with Garman. I'm that guy that I'm following route. And I always end up with more mileage. So I, up going out to HaCo. And I follow this route and it had so much more climbing, even though I am not built like a climber. I love as these challenging routes. I really love taking on routes to have as much climbing as possible. And it, even though it takes me all stinking day and HaCo has that, you know, for Texas as one of these guess that you get so much climbing and I absolutely fell in love with it. And Heico has it's heart packed. And it has a lot of beautiful canopy areas, you know, tree canopy, tree areas. It has lots of water crossings. It has. I mean, it's just a really diverse terrain. And even if it were to rain, it's so hard packed. It's it's got a lot of smooth areas. There's really nothing rough or nasty about it. It, and. Even, even if it rains it's, it's not an area that, that you get much mu much more than a couple inches of mud. So it's not like, like Mid-South where you have, you know, six inches, 12 inches of mud in your foot, you know, is, you know, foot into the mud. For example, last year in may it did run, it did rain. And so yeah, people got my, but you're talking, you know, an inch of mud, two inches of mud. Maybe two and a half. So it's not a situation that you're just bogged down and you, you have to walk, you, you can ride through it. So it it's really a, an area that you can ride it all year long rain or shine. And it was just perfect. And. I said, this is it. We're gonna do it here. So, you know, we had last year, we had three routes this year. We have four routes. Last year we had a 30 mile or a 60 and 150 something. And after our survey of the event, it was a very positive survey folks, which is absolutely thrilled with the event. But what kept coming up was have a 100 mile. I said, all right. So for 2022, we have a hundred mile as well. And it just filled up immediately. So folks really wanted a hundred. Not everybody wants to do 150 something miles, but they want it more than a 60 mile. So the hundred mile, you know, I never even thought of it, you know? So you learn, I've learned a lot, you know, I thought, [00:18:03] CraigDalton.: feel, I feel like I'm in that category where 150 mile, maybe I can muster that up once a year, if I'm lucky, but a hundred keeps me honest. I need to train for that pretty well. But I, I believe in my heart, like I can always uncork a hundred miles if I'm like relatively fit. [00:18:20] FabianSerralta: Yeah. And you know, and the hundred mile it is, it's a legit route. I mean, it's, it's over 5,000 feet of climbing and it's really a beautiful route and it, and it really incorporates all the hard climbs that are in the 150 something mile. And [00:18:33] CraigDalton.: what I, one of the things I always wanna unpack with event organizers, because I think it is a challenge depending on where you are, is okay. So you you've decided on HaCo as a, as a great riding location. But there are also logistics and permits and all kinds of things. You need to go through granted in a rural community. Maybe those are less than a more populous community, but you still need to do that. So what was that process? What was that process like for you? [00:18:59] FabianSerralta: Really easy, you know, what, what I have learned with I, you know, this is for me as a hobby and HaCo has, you know, it's really been incredibly easy. They they've really take care of all that for me. And I went in there with, you know, the understanding that, look, I'm doing this to help the volunteer fire departments. If you're willing to help me, I'm willing to do it. [00:19:17] CraigDalton.: And was that perspective, something that was already in your head. Hey, I wanna have a charitable component to the event I put on. [00:19:24] FabianSerralta: Yes. Yes. And, and if you're willing to work with me and, and take care of these things, I'm willing to do it. If you're not willing to work with me, I'm outta here. I just, you know, it's, it's one of those things that I, I don't have the time to mess with all that stuff. So if the town is willing, then I'm willing, I, if they want to put me through all these hoops and things and, and, and, and barricades and all these. All this red tape, I'm just, I'm outta here because I just don't have time for it. You know, I've got four kids and two other businesses to run. And so I was really upfront and they were really honest about it. And they just, they facilitated everything that I needed. They, they provide all the things that I needed. They provide law enforcement, they provide crowd control and barricades and they provide everything. The same thing with Pueblo, you know, they're. [00:20:09] CraigDalton.: you know, hike in the original gravel locus event. Sorry to interrupt you there for it. It took off through the gravel cycling community as a event option incredibly quickly. And there was a couple things that. Kind of at least caught my eye right off the bat, which were one was correct me if I'm wrong here. But it seemed like the registration was entirely a donation based model, which was unheard of. And two for a first year event, you had all these top pros saying I'm gonna be there. [00:20:40] FabianSerralta: Yes. [00:20:40] CraigDalton.: How did both of those things happen? [00:20:43] FabianSerralta: well, you know, it was, it was out of really, so the event was initially gonna take place in 2020 in November and I canceled it cuz of COVID, you know, so I had the Greenlight from, from HaCo, but I canceled it because of COVID. My basically, you know, I have absolutely zero connections in the bike industry. And what I tried to do was try and get the bike industry board. And the only way that I could do that was reaching out to them via Instagram and Facebook. And that really didn't really work. I couldn't get anybody to, you know, return any messages or anything, despite the amount of money that I spent on bikes. You know, I have the the record of my local bike shop for spending the most money on bicycles every year. just absurd. despite all that I couldn't get a response from anybody. So I said, you know what? I'm not gonna let that discourage me. I'm gonna have this event with, or without the bike industry. And so I said, I'll, I'll fund it. I'll do it myself, cuz I I'm gonna have this event. And this event is gonna have the component of the pros and the component of the beginners. And there is not there. Isn't gonna be a cutoff. So if folks are gonna train for the geo one fifty, a hundred fifty something miles with over 8,000 feet of elevation gain. I'm not gonna yank 'em off the course. They're gonna be out there as long as they want to be. And if they want to give up, they're gonna give up, but it's not gonna be, I'm not gonna take it away from anybody. I'm not gonna be that person. That's gonna say, Hey, you know what? You didn't make the cut off by 10 minutes or an hour or two hours. And you're off the course. Because I'm always that person, who's the last one. And you know, when I was at Ted's event in Vermont last year, I didn't make the cutoff. And when they came up to me and they, Hey, look, you, you didn't make the cutoff. I said, , we're gonna have a fricking fight. . And they said call Ted. And they did. They called Ted and it's like, leave him alone. He's fine. I I'll take care of, I'll wait for him. And Ted did. And he understands, he, he waited out there for me and I didn't make the cutoff by over an hour. And he was out there in the rain, in the cold waiting for me, everybody was gone. The whole thing was shut down. And he followed me for like the last 15 miles, cuz it was pouring rain. It was cold, but you know, he, he let me finish the, the event and to me, that's what being inclusive and, and finishing and, and you know what it's about. So to, Hey said, I'm gonna have this event regardless. So in 20 for 2021, how I was able to. Do what I did is with, like you mentioned earlier, this, this donation thing I said, you know what? Let's just, I've gotta get the attention of, of folks. Cause I don't have the support of the bike industry and I don't have name recognition and I certainly don't have, you know, experience. I've never done this before. So how do we capture attention? We're gonna do this for free. It's gonna be a free event. And, and first thing I said, all right, this event is gonna have 1200 free registrations. And what you're gonna do is it's up to you to make a donation, a direct donation to the volunteer fire department. Most folks are used to paying a hundred, something to hundred dollars or more for an event like of this caliber. Remember you have all these age stations you have. So you're getting 20 something pros. Top level pros from around the world. You're getting over five age stations, fully S stocked, 20 something, sag vehicles, portable bathrooms at all. Age stations. You're getting a draw string swag bag. Really nice. You're getting with zipper. You're getting T really nice. T-shirts you're getting purest water bottles from specialized. You're get all this swag, all this stuff for free. So, whether you give the fire station a dollar or $0, you're gonna get everything for free. So it was an honor system. And really, I think that showed people that it, Hey, this is an event that has Lawrence 10, Dan Ted king, Peter TNA, Allison Terick Jess, Sarah. You know, Colin, Strickland, you know, all these names that I, that are all of 'em are gonna be at at Unbound, all of 'em are, are all these huge events and there's no cutoff. There's all of this support that you don't get at these big events, you know, as far as aid stations and it is entirely up to me to decide how much I'm willing to pay with what I think it's worth well out of those 1200 free registrations. Less than 400 people donated anything. So that was a bit of a shock, but we still had the event [00:25:11] CraigDalton.: Yeah. And, and just to unpack that a little bit, you know, quite disappointing, obviously, that just like sort of the percentage of people that actually donated and to, to put a finer point on it, like, as you describe all the things that one would get for participating, you're talking about a hundred dollars worth of. Effort per rider to give them nutrition, to provide them porta potties, to give them swag, all those things. So it's a, it's a big proposition that I think often gets lost and and you put it in the proper context in that without someone supporting you without a, you know, a nutrition sponsor coming in and dumping. Tens of thousands of dollars worth of product on your tables. Like that's coming outta your pocket as the race organizer. And there's no way around that. So pretty incredible effort to get it off the ground. And sorry to hear that the donations were not as great as you wanted them to be, but with those donations, something great. Did material realize for the fire department. [00:26:12] FabianSerralta: Yeah. And. That that side didn't happen the way that it had. I had hope, but it, we don't really even think about it because we, at the end of the day, there were so many articles written about the event that I never expected in a million years. I honestly, I never thought that GCN would write about it. That basically magazine would write about it. That cycling news, the Velo news, all these publications wrote about it. You know, it was listed as a, as a basically magazine listed as a. Top 20 bucket list event. You, it was mentioned like 17 times in VLO news. GCN mentioned it cycling news had articles about it. These are all things that I never even consider would happen. [00:26:55] CraigDalton.: Yeah. it was absolutely incredible to reach the brand and the event got in that first year. [00:27:00] FabianSerralta: We smoking great deal on the, the fire. We were still able to buy it with the money that we raised. We, you know, we still had great registration numbers we had. And then for 2022, you know, we have over 1500 people registered and we raised enough to build a bigger fire station. So [00:27:20] CraigDalton.: That's [00:27:20] FabianSerralta: while maybe, you know, less than 400 people register out of 1200, it doesn't matter to me because cuz we still accomplished everything that we set out to do. We still got more numbers than I ever thought were gonna happen. And as a result, we, we have interest from other cities that are contacting me that wanna have events. So, you know, I, I was several cities reached out to me. Hey, can, can you do the same at our city? And it is, you know, I've had to turn down cities cuz it's just too much. You know, I've had two, two other Texas cities that I asked me to host events at their cities. And I unfortunately I've had to say no because I just don't have the time [00:27:59] CraigDalton.: Yeah. Yeah. [00:28:00] FabianSerralta: Pueblo. Against my better judgment. I was like, sure. It's you know, the, the Pueblo story. I really, really I, I couldn't say no, it's just, they they've, they've been wanting to do an event there and they even they were bidding to try and get an event and they, they lost the bidding most cities that want to do something like this. They're paying promoters to do the events there. That's one of my rules. I will not charge a city to host any event, cuz it goes completely against what I'm trying to do. So what I'm trying to do is bring money to the city and build and bring equipment and funding to the volunteer fire departments. So why would I be charging them money to host events [00:28:39] CraigDalton.: Yeah, it's super, it's super interesting. When you look at the economics of events, just events in general, how much they cost, but the economic impact to these rural communities, which in your case is very specific to raising money for these volunteer fire departments. But even beyond that, the, you know, the amount of meals purchased the amount of hotel nights, cetera. Like it's it, it has a significant impact. and, I'm super interested to get into your second event in Pueblo and learn how that happened. But one more question, just outta my own curiosity, how did you end up getting those 20 to 30 pros to come to a first year event? [00:29:15] FabianSerralta: I will see the credit to that goes to Ted king because I reached out to several pros. First was him. Via Instagram. Again, I don't have any connections or I don't really know anybody. And he was the only person that responded. And I wrote him this long thing through Instagram, direct messaging through Instagram. And I explained to him, look, this is what I'm trying to do. I'm I'm gonna have this event and it's gonna be free to everybody and gonna be up to them to donate. And this is all that they're gonna get, and this is all I'm gonna give them. And this is what I'm trying to buy a fire truck for the fire department. And I'm trying to get this many pros and he, he rides back. He's like, are you nuts? and he finally calls me. He's like, are you nuts? He's like, I'm so intrigued by this. And. [00:30:04] CraigDalton.: The idea, the idea was so crazy. He had to call and talk to the guy behind [00:30:08] FabianSerralta: That's exactly what he said. He's like, this is absolutely insane in this, but I'm so like intrigued by this and he's like, are you really gonna do this? And I said, oh yeah, I'm really gonna do this. And he's like, you know, this is how stuff gets done. You know, when, when people just take a chain and, and, and do something completely outside of the box. And, and he said, can I have two weeks to think about this? And I said, absolutely just take your time. And and he's, and he did two weeks later, he calls me back and I'm like huge fan of Ted. And, and I remember watching him in the tour de France and all that, and sure enough, two weeks later he calls me back. He's like, all right, I'm in. And. Do you have a website? No. He's like, you need a website and then he's like, what are you gonna do for registration? I don't know. I mean, he just went down this list. He's like, you gotta have registration, you gotta have this. And, and then and then he says to me and your social media post suck [00:31:11] CraigDalton.: suck [00:31:12] FabianSerralta: and he is, starts helping me, you know, he starts Giving me so much guidance and stuff like that. And he is like, and how are you gonna get ahold of all these other riders? Cause I give 'em a list. Like I want to get ahold of, of all these other riders. And he is like, I don't know. He says that, you know, let me help you. So he started really vouching for me and, and contacting them. And. And then he gave me a lot of advice. He says, you know, don't, don't do a don't, don't give money, don't do a purse, don't do this and don't do that. And, and you're gonna find that the folks that believe in what you're trying to do are gonna jump on board. And, and that's how it happened. He just, the folks that came forward are, are folks that care about what I was trying to do. And really wanted to be a part of something totally different. That was more about giving back. To communities and, and, and not so much about, you know, a big corporate event, it was more of a Grasso type thing. They, they were just interested and a lot of 'em have their own events that are grassroots focused, you know, Ted and Jess, Sarah, Sam Boardman, Laura King, and Ted, you know, all of them Lawrence Tanem has his own events in the Netherlands. Peter has his own event. So all of there's a connection amongst all of us. That have to do with our own little small events that are, they're not corporate they're just small community type events. [00:32:32] CraigDalton.: Yeah. Yeah. Amazing, amazing [00:32:34] FabianSerralta: then what would I do in return? You know, I help with the hotels and things like that and meals and stuff like, you know, that's how, what we do, you know, they, they ride for a living that's, that's, that's how they make a living. So you certainly, you have to help out in some way, you know, and you know, when with HCO helps me out with the hotel cost, so I provide them with a room and, and stuff like that. [00:32:54] CraigDalton.: Gotcha. Interesting. So now let's shift gears to Pueblo. I know you had mentioned you had a number of rural communities. See what you had done and reach out to you, but what was it about going to Pueblo in a state that you don't and you don't have property at this point? What was it about the Pueblo opportunity that said, Hey, this is the right next step for the gravel locus event team. [00:33:16] FabianSerralta: Well, I, I found out that they helped, they they've been wanting to have a gravel rent and I found out that. They had been bidding on, on, on having an event hosted there and, and they lost somebody else got the bid and that kind of bugged me. It bugged me, it bugs me that these small towns that are hurting and economically, and they're trying to bring business and they're trying to bring funds to their towns that they're, that there. Trying to get events by paying promoters and paying corporations money or offering to pay money. So that events are being hosted there. And to me, that it just doesn't make sense because if you're doing it for, for, as a business, you're making money. If, if you're hosting an event somewhere, you're gonna make money from registration, you're gonna make money from cells of, of, of, you know, shirts and, and you're making money from. Vendors and, and the bike industry's paying you per and, and, and all this stuff that I, I have, I don't have access to cause I don't have any, you know, I don't have the bike industry behind me and you know, or any of that, but you know, the, the lifetime events and the big corporate events have all of that, you know, you know, all those, you know, shaman and spa and all those companies that are, there are not there for free, you know, let's just be honest, you know, I was born at night, but not last night. So, And then to have these small towns that are struggling financially and have high unemployment paying significant amount of money for, for them to host the event there, to me that that just seemed wrong. And as a result, they, they were outted by another town and they didn't get the event. And I learned about this and I said, all right, we'll do the event there. So I met with the city I met with the mayor. And one of the questions were, well, how much are you gonna charge us to do the event here? I said zero. And I said, if I ever ask you for money to do an event here, kick me in your ass, please. I said, that's not. That goes completely against what I'm trying to do. I said, the reason I'm I'm doing these events is to bring funds to cities that are struggling financially, but also to bring funds to the volunteer fire department, because. Guess who we call when we fall and wipe out and bust our asses out, riding gravel, volunteer, fire departments, guess who's out there. That's, who's out there. You know, when we're out there riding gravel that, and all of us know this it's volunteer fire departments. We're out in the middle of nowhere and it's a volunteer fire department. It out there charge and really taking care of hundreds of, of miles. That they're covering and that's who we rely on. So if we're gonna support a, a department as a gravel community, I mean, I would think that I, you know, logically we're gonna support the volunteer fire departments in those areas that we ride. So it's not that I'm Mr. Nice or anything like that. It's just logical that we're gonna support the very same people that come to help us. And it's the volunteer fire departments in those areas that we ride. So. And I think they, they appreciate that, you know, so to charge them, it doesn't make sense. I don't think it's fair, but they do help. You know, they provide, like we said earlier, they help me with the permitting and they help me with law enforcement and they help me with barricades and things like that. So they, they do help. It's not like they're doing nothing. So it's, it becomes a, a community, a true community involved event. And we, we get that. We throw that word around all the time, community, this community, that, but when you really dig into it, poor community is paying a few hundred thousand dollars. And it's really no longer a community. Now we're talking about a service and a fees and stuff like that. But in, in our case, you know, HaCo provides all this help and volume and stuff like that. The same with Pueblo. That's. [00:37:03] CraigDalton.: think that's, you know, it's, it's in, I think it's refreshing to take that expansive view of community cuz oftentimes the gravel community, those words are thrown around a lot. Generally implying your fellow athletes, the fellow people out there riding with you. But it, it really is in these rural communities. It's the people of the community that are coming out, coming out, whether they're, you know, Manning the registration booth or Manning an aid station, or, you know, opening their doors and giving you a glass of water. If you're stuck out there somewhere like that, that really is the breadth of the community that gravel does touch in these town. [00:37:38] FabianSerralta: Oh, I mean, you better believe it. If they don't help me, I'm not doing it. mean, there's no way I, I would do it. It it's just, you know, I wouldn't do it. And they understand that and, and, and I'm really open about that. It's like, I'm not charging anything, but you gotta help me. You know, when I met with the fire department, it's like, you all gotta help me. There's, you know, I don't have, this is, you know, and I, and I say this, you know, like, My social media, it's one Cuban and an iPhone. You know, I don't have a, a, a, a crew or anything like that. It's, you know what you see on social media? It's Fabian, you know, one Cuban and an iPhone. I don't have employees. I don't have anything. It's just myself. So I, I will take all the help that I can get. And you don't need an entire staff to, to do anything like this, but you do need, you know, help and, and volunteers and, and Pueblo understands that. And HaCo understands that, you know, I'm looking at another state right now and we're looking at a third event and they understand the same thing that, Hey, we need, we want to have event grab a locus type event. We don't have, you know, a few hundred thousand dollars to. Give a promoter to bring the event here, but we do have willing bodies and people that are willing to, you know, help and, and volunteer. And, and that's the model. And, but we also like having 20 something pros and we like having the, that racing component. But yet we also want to have an environment that the Fabians that are gonna come in last know that they can train for this. And it's not gonna be taken away. They're not gonna be turned around and yanked off the course because to me, that's, I can't imagine training for something for a year. And, you know, not being a Ted king or, or, or a Allison Terick or Jess, Sarah, I'm not those folks. You know, I can't imagine training and having my family behind me and all of that support and, and the struggles of having to work other jobs and then being yanked off the course until, Hey, you know, you can't finish you because you didn't make it by 30 minutes or an hour or two hours. I cannot imagine what that feels like. And to me that just, that that's not inclusive, that that's just telling somebody you're not good enough and turn around. I, I, to me, there's no explanation that you can give me that tells me that's rational, reasonable, not even safety. What do you mean safety? There's, there's nothing. If it's a issue then guess what? You know, the amount of money that these events make, then you plant a fricking sag vehicle behind those folks to follow them till the end. What does that cost? I mean, lemme know I'll pay for it. What's the big deal. You know, and that's what we do at gravel locus. Last year, we had a vehicle sag vehicle to follow five folks, cuz it was dark. They didn't have lights. I said, well you freaking follow them. Follow 'em all the way to the end. You're gonna be their light because. Again, why would we yank somebody off the course? Now, if they want to quit, it's on them. If they want to throw in the towel, it's on them. But, and, and then the other thing that we have with our pros and, and, and they're more, and you, we don't even have to ask em and it's like, Hey, you know, make yourself available to the, to the folks, to the regular folks. And I do, there are so many selfies out there with Ted and, and kids and, and Pete. Lawrence. It's just super cool and funny as hell. Allie Terick and Jess, you know, all these young girls and, and folks that got to meet them personally. And they were out there available to all these folks, which, you know, it's, those are memories and things that you just, you know, all their fans get to meet them. [00:41:16] CraigDalton.: A hundred percent. So I guess we, you know, I feel like we've unpacked pretty completely what the gravel Locos, eco ethos is and what the experience is gonna be like on the Pueblo side. What is that terrain like? I've driven through Pueblo, but I've never, I've never put rubber on the its and trails there. What, what do you expect the courses to be like in Pueblo? [00:41:36] FabianSerralta: Oh, my gosh, it's silky smooth, but you know, it's, it's, it's so beautiful. I got to ride 23, 24 miles the other day. So we, we, we rode from the fire station. We did a loop and it was about 24 miles and about 1400 feet of elevation gain. The the main route we call it the, the GL one 50, that's just our, our, our, the, the big route is always gonna be called the gravel locals one 50, but it's really 169 miles. So you get, you get a little extra, but it's, you know, right with GPS says it's over 13,000 miles, but as we know, it's always under, it's always more than. [00:42:14] CraigDalton.: feet of climbing. [00:42:15] FabianSerralta: So I would expect just about 14,000 feet of elevation gain and, you know, it's it's for me coming from Texas, it was a little, a little harder cuz there there's that whole, you know, the altitude, but it is just so smooth and so nice. It's you know, it's hard packed also. It's it's smoother. The gravel there is it. It's just hard to explain. It's just, it's beautiful. [00:42:40] CraigDalton.: So do you imagine this being the type of event that larger groups can stay together and benefit from drafting off one another and things like that? Just given the type of terrain you're on. [00:42:48] FabianSerralta: Yeah. Yeah. It's just, I mean, it, it's hard to explain. I mean, there it's, I was, I was telling them over there, it's like, it's hard and it's definitely challenging, but you're looking at mountains and you're looking at, at all this beautiful terrain, it's really, you know, I was, I was exhausted. Again, I'm not, I'm not anybody who's in the kind of shape and certainly I've, I've gained. I, I was telling them the other day, I was like, you know, there was the COVID 19 pounds. I, I managed to, you know, I went from riding a ton of mileage every year. And when I started gravel Locos, I've gained 50 pounds in, in two years of not riding and running two businesses and gravel Loco. So. I've gotta find a way to get these 50 pounds off. So yeah, going downhill was great over there, but going up was tough, but just a scenery is just so, so interesting. [00:43:37] CraigDalton.: and are the courses punctuated by like a significant climbs? Like in terms of like, oh, you're gonna be climbing for an hour at a time. Like [00:43:44] FabianSerralta: know, gradual they're gradual climbs. There was nothing that like in HCO, you get these punchy climbs over. There's more gradual. So. You know, you can get away with with less big gears in HighCo. I tend to run bigger gears over there. You know, I didn't get in any of my big gears, you know, I, I run a 10 52 in a rear Ram and I, I wasn't using it over there because they're, they're more of the gradual climb. You kind of get into a rhythm and they're kind of, I prefer that to be honest, that you just kind of get into this groove and you get into this rhythm. Whereas high code has these. You know, we have some, some climbs that are 22% 23, and they're pretty punchy. And we have these three climbs that are back to back that we call the three bees. You can decide whatever you want to call those bees, but bitches they're rough. And then and man, those kick up into the 22, you 23, 20 4%. and it's, you know, they're kind of loose rocks and you just kind of gotta lean forward and you gotta get that big granny gear and just work your way up. Whereas in Pueblo, you don't have, I didn't experience anything like that. And, and most of what's out there is these long gradual climbs that you kind of get into a rhythm. So I prefer the, the type of climbing that they have out there in Pueblo, but they're both very, very different, very different type of riding. [00:45:05] CraigDalton.: And would you change your, your tire width from HaCo to Pueblo? Would you do [00:45:10] FabianSerralta: I think that you can. I think that at, at both, you can get away with smaller diameter tires. So I've done HaCo with 35 centimeter tires. I've done HaCo with 47. I think that in Pueblo again, we rode Pueblo last week where it had been snowing. and it was muddy, but even then, you know, I wrote it with 45 centimeters and there was folks out there running it, riding it with 30 eights and they still did just fine, but the, the Pueblo gravel is much smoother than the HaCo. And I, I can see some folks getting away with when it's dry, especially over getting away with, you know, 30 fives, [00:45:50] CraigDalton.: Yeah. [00:45:50] FabianSerralta: somes. I. [00:45:52] CraigDalton.: It's funny. It's so it's so counterintuitive to me to, and S B T gravels the same way I remember getting like my arm twisted to run 30 eights, and I was like, there's, there's no way, like I'm going to Colorado. And at home, I'm all about the 40 sevens here in Marin county, cuz it's so Rocky and but sure enough, like I definit could have ridden that as they call it champagne, gravel in Colorado on 38 with absolutely no issue. So it, it, it's quite fascinating to me and. One of the things I. [00:46:20] FabianSerralta: racers recently that are 40 threes. I haven't put 'em on yet, but they're kind of, you know, they're not slicks, but they're not Nobbies and I can't wait to try something like that. They're 40 threes. And normally in HighCo I run 40 fives or 40 sevens, but I'm looking forward to running those 43 as a whole new tire I've never used for, but I can certainly see a. And in Pueblo getting away with 38, even, even a slick or a semi slick. [00:46:45] CraigDalton.: Yeah. so, so interesting. It's obviously, it's like, it's a never ending debate and discussion about which tires to use. And I remember reading, you know, where the, when we're recording, it's the weekend of Mid-South and you know, there's a lot of people. Twisting their arms and, and twist getting all twisted inside about what tires are gonna run tomorrow or today. Excuse me. On race day at Mid-South. So always fun and appreciate the insight there. So for people looking to find out more information about the events, where can they find information about gravel locus? Why don't you tell us the, the website and the dates of the events this year? [00:47:19] FabianSerralta: So HaCo is May 14th. And Pueblo is October the first you can sign up on our website, which is www dot gravel, locos.bike. Just make sure you click the link for one or the other, or if you wanna do both I've left. He still opened. But we've got a or 1500. I haven't decided where I want to shut it down because HaCo is open to having more Pueblo is still open as well. but just all the information is on there. There's four routes for each. So there's the gravel locals, 30, the 60, the hundred. And the one 50 information about the routes is on both of them. The cause for each of the events is always gonna be a volunteer fire department. Pretty much everything you need to know about it is on the website. You'll find all the different pros that are gonna be there. We've got more pros to add to the website every day you get, you know, we get new pros that wanna come on board. As far as our sponsors there, aren't not many of them, but the, the ones that are on there, you'll find out that these are folks that are very, have been very loyal since the beginning. And if somebody wants to be a sponsor, they're more welcome. Welcome to send us a message or email us, but. We're pretty much self supported and really it's just a community thing. And, and it's really, this whole entire thing has been made possible by, by the towns and, and little businesses within the towns. Very small involvement from the bike industry. It's been a, a community thing to be on. Oh, just kind of how it worked out. [00:48:42] CraigDalton.: Amazing. Thank you so much. Thank you so much for putting so much of your heart out. there and, and making a making events that you wanted to see happen in the world. And I think it's such a sort of beneficial place for the gravel cycling community to have event organizers with that orientation and From this conversation in our earlier conversations. I know how much of yourself, both personally and financially you put on the line to create this event series. So again, for, from, for me, thank you for doing that and exciting to hear that Pueblo is going off this year and exciting to hear that yet another community has come to you and talking about like, how do we have grow Ava three. [00:49:19] FabianSerralta: Yes, we we'll release that soon. I'll let you know. . [00:49:23] CraigDalton.: Awesome. Thanks for your time, Fabian. [00:49:25] FabianSerralta: you. Thank you. So. [00:49:27] Craig Dalton: That's going to do it for this week's edition of the gravel ride podcast. Huge. Thanks to Fabion, not only for joining us on the show, but for everything he's doing around the gravel Locos series, I think he's got his heart in the right place and I've no doubt. These are some of the best gravel events out there to attend. Big. Thanks. Also to our friends at hammerhead for sponsoring the show member, you can get a free custom color kit and premium water bottle with the purchase of the new hammerhead kuru two computer, simply go to hammerhead.io and use the promo code. The gravel ride. If you are interested in supporting the show, ratings and reviews are hugely helpful as is sharing the show with other gravel cyclists. If you're able to support the show financially, simply visit buy me a coffee.com/the gravel ride. Until next time. Here's to finding some dirt onto your wheels  

Jeff was back up in Georgia again this week (who can blame him), but through the wonders of modern technology, he was able to hang out with Capt. Kevin and Capt. Kirk to talk about the INCREDIBLE weather that greeted them this morning.  It was a beautiful day, and provided the perfect backdrop to talk about getting ready for bow season, as well as talking about the upcoming mullet run. Here's your L.V. Hiers gear tip of the week: Jeff says the Boat Gadget 10-in-1 Boat Tool is the ONLY tool he needs on his boat… It literally does everything! Stop by Hagan Coastal Outfitters inside Hagan Ace Hardware of Mandarin, Green Cove Springs and Yulee to get yours!  For the HCO location near you go to: www.hagancoastaloutfitters.com Here's your Ring Power CAT tip of the week: Dove season opens in Georgia this weekend! Get yourself a MOJO dove decoy HERE Here's your Kirbyco Builders Inc. cooking tip of the week: Sweet Potato Soufflé 1 lb steamed sweet potatoes 1 stick of butter 3 eggs 3/4 cup sugar 3 tbs flour(any) 2 tbs vanilla extract 1 teaspoon baking powder Blend thoroughly in blender until it's a thick soup Pour in a greased 1.5 qt casserole bowl Bake at 350 (45 mins) (Top with marshmallows optional but recommended!)

In the wake of the recent IPO, we interview three early members of the Airbnb China team on how Airbnb found and stuck to its core advantages against an array of local competitors. And how it managed to emerge as a rare Western tech startup success story in China. Guest lineup: Robert Hao, Co-Founder and COO of Hype, Ltd., Director of Business Operations and Strategy, HCO, Head of Supply at Airbnb 2012-2017 Bryan Huang, Founder at Annabai Technology, IT Manager at Airbnb 2015-2018 Ziyang David Fan, Head of Digital Trade at World Economic Forum, Senior Legal Counsel at Airbnb 2015-2017

In the wake of the recent IPO, we interview three early members of the Airbnb China team on how Airbnb found and stuck to its core advantages against an array of local competitors. And how it managed to emerge as a rare Western tech startup success story in China.Guest lineup: Robert Hao, Co-Founder and COO of Hype, Ltd.; Director of Business Operations and Strategy, HCO, Head of Supply at Airbnb 2012-2017 Bryan Huang, Founder at Annabai Technology; IT Manager at Airbnb 2015-2018 Ziyang David Fan, Head of Digital Trade at World Economic Forum; Senior Legal Counsel at Airbnb 2015-2017

¿Cómo tengo que alimentarme durante un 10k, una media maratón o una maratón? ¿Cómo debo hidratarme y cada cuanto tiempo tengo que tomarme un gel? ¿Es posible mejorar la capacidad de absorción de HCO de nuestro cuerpo¿ ¿Afecta el nivel de hidratación a la absorción de los HCO en carrera? ¿Qué podemos hacer con nuestra nutrición para afrontar mejor "El Muro"? Hoy respondemos a estas preguntas y muchas otras más con la ayuda de Carlos Aznar, triatleta, entrenador y nutricionista. 👉🏽 Puedes encontrar a Carlos en rrss como @aznargallego 🏃🏾‍♀️ Si necesitas cualquiera de los nutrientes y suplementos mencionados en el episodio, puedes encontrarlos en www.feedmiles.com 🎁 10% de descuento con el código RUBENESPINOSAPT para compras superiores a 30€

¿Cómo tengo que alimentarme durante un 10k, una media maratón o una maratón? ¿Cómo debo hidratarme y cada cuanto tiempo tengo que tomarme un gel? ¿Es posible mejorar la capacidad de absorción de HCO de nuestro cuerpo¿ ¿Afecta el nivel de hidratación a la absorción de los HCO en carrera? ¿Qué podemos hacer con nuestra nutrición para afrontar mejor "El Muro"? Hoy respondemos a estas preguntas y muchas otras más con la ayuda de Carlos Aznar, triatleta, entrenador y nutricionista. 👉🏽 Puedes encontrar a Carlos en rrss como @aznargallego 🏃🏾‍♀️ Si necesitas cualquiera de los nutrientes y suplementos mencionados en el episodio, puedes encontrarlos en www.feedmiles.com 🎁 10% de descuento con el código RUBENESPINOSAPT para compras superiores a 30€

We're back after a brief hiatus and get in to the COVID-19 issues: masks, Victoria's lockdown, the hydroxychloroquine debate, and Big Brother spying on us to make sure we don't get too close. We also discuss the explosion in the port of Beirut and American mob stories. Simon then introduces us to Scott, who he became friends with through a common love of the music of Bruce Springsteen. Scott has written an inspiring book, "Head Strong: How a Broken Neck Strengthened My Spirit", about his amazing life story so far. Watch Scott's story here: https://www.youtube.com/watch?v=-Ng6Q_Oxad4 and get his book here: https://www.scottwfedor.com. Follow us on Facebook at www.facebook.com/ChroniclesOnTheFly, on Instagram at www.instagram.com/chroniclesontheflypod, and on Twitter at www.twitter.com/ChroniclesOTF

“Stop met talenten in de vuilbak te gooien omwille van afkomst.”Gerenommeerd comedian Nigel Williams vertelt over zijn moeilijke jeugdjaren in Engeland en hoe hij er zich letterlijk en figuurlijk uit heeft kunnen vechten.

Si hay algo que todo el mundo busca es accelerar su metabolismo. Y aunque técnicamente no se puede, si que hay algunos trucos para favorecer el aumento de la masa magra o muscular que es la que marca el metabolismo basal. Además hay otros factores que también pueden ayudar a ese incremento:1. Entrenamientos HIIT. Este tipo de entrenamiento generará tal estrés en tu cuerpo que hasta 72h seguirás estando activo.2. Levantar pesas. La musculatura es un órgano y por ello, activarla con una tensión muscular suficiente, provocará que tu cuerpo produzca unas hormonas anabólicas que acelerarán tu metabolismo hasta 39h después del entrenamiento.3. Asegurar la ración proteica. Al digerir los alimentos necesitamos energía para poder realizar el proceso que los descompone. Las proteínas, al contener nitrógeno son mas difíciles de descomponer y quemaremos un 25% de las kcal que nos aportan para poder hacerlo. Un número considerable teniendo en cuenta la energía que necesitamos para el resto de macros. (2% para grasas y 5% para HCO)4. Beber más agua. Estar correctamente hidratado ayudará a que tus órganos regulen adecuadamente su funcionamiento y esto producirá menores probabilidades de que tu metabolismo vaya más lento.5. Minimizar el estrés. Nuestro cuerpo sigue viviendo en el paleolítico y el estrés significa "no encuentro comida". Por lo tanto, tenderá a ralentizar el metabolismo para sobrevivir. 30' de meditación al día reducirán tus niveles de cortisol y te ayudará a aumentar tu metabolismo.

Si hay algo que todo el mundo busca es accelerar su metabolismo. Y aunque técnicamente no se puede, si que hay algunos trucos para favorecer el aumento de la masa magra o muscular que es la que marca el metabolismo basal. Además hay otros factores que también pueden ayudar a ese incremento:1. Entrenamientos HIIT. Este tipo de entrenamiento generará tal estrés en tu cuerpo que hasta 72h seguirás estando activo.2. Levantar pesas. La musculatura es un órgano y por ello, activarla con una tensión muscular suficiente, provocará que tu cuerpo produzca unas hormonas anabólicas que acelerarán tu metabolismo hasta 39h después del entrenamiento.3. Asegurar la ración proteica. Al digerir los alimentos necesitamos energía para poder realizar el proceso que los descompone. Las proteínas, al contener nitrógeno son mas difíciles de descomponer y quemaremos un 25% de las kcal que nos aportan para poder hacerlo. Un número considerable teniendo en cuenta la energía que necesitamos para el resto de macros. (2% para grasas y 5% para HCO)4. Beber más agua. Estar correctamente hidratado ayudará a que tus órganos regulen adecuadamente su funcionamiento y esto producirá menores probabilidades de que tu metabolismo vaya más lento.5. Minimizar el estrés. Nuestro cuerpo sigue viviendo en el paleolítico y el estrés significa "no encuentro comida". Por lo tanto, tenderá a ralentizar el metabolismo para sobrevivir. 30' de meditación al día reducirán tus niveles de cortisol y te ayudará a aumentar tu metabolismo.

I read out an interesting HCO executive letter by Hubbard. It mentions some of the attacks the church was getting in the 60s and that the church defeated them. Why would the government have stopped trying to take down Scientology after the failed attempts in the 60s? I suggest that they continued trying and eventually succeeded in the 80s. Andy's Scientology KRs are secret closed door briefings on information about Scientology and the world that you need to know. Hosted by Andy Nolch the space cowboy. Available on most podcast apps & Youtube. Remember get up that bridge!

"Wel lastig dat het voor 4.000 mensen moet gebeuren..." Rapper Kim Nzita Vangu, alias Pasi, vertelt over levenslang leren en hoe het noodlot je toch in de juiste richting kan duwen.

"Sinds hij gepasseerd is, hebben we al onze klinken moeten vervangen." Scenarist en animator Johan Bolsens geeft een unieke kijk in een asielcentrum en vertelt hoe één persoon hem altijd is bijgebleven.

"Ik zie die eend, samen met die steen, naar onder gaan. En ik denk: Shit!" Schrijver, zanger en impro acteur Jules Jordens vertelt over een avontuurlijke jongensachtige reis en hoe je in moeilijke situaties op je echte vrienden kan rekenen.

"Ik wil de leerkracht zijn die ik niet heb gehad."Leerkracht en spoken word artieste Sascha Reunes vertelt over hoe ze vroeger op de middelbare school ongehoord was maar nu op haar beurt de bijna onbereikbare jeugd probeert te bereiken.

"Ooit vroegen twee vrienden aan me wat liefde was... en dit verhaal kwam in me op." Danstherapeute, theatermaakster en danseres Kristien Maus vertelt hoe eerlijk dansen soms levens kan redden.

Suzanne Horstman is the director of Home and Community Options in Winona MN, providing services to developmentally disabled citizens. She’s heading up an organization doing amazing work, and joins Brett to talk about the mission and the passions behind her 27 years of working with HCO.

Suzanne Horstman is the director of Home and Community Options in Winona MN, providing services to developmentally disabled citizens. She's heading up an organization doing amazing work, and joins Brett to talk about the mission and the passions behind her 27 years of working with HCO.

Brendan and Bryan are back to talk Irish culture, purple Guinness, long wet winter women, Conor McGregor/Paulie Malignaggi rumors, guy code, street fights, O.J. Simpson, Trump, freaky girls, HCO's and much more.See omnystudio.com/listener for privacy information.

Brendan and Bryan are back to talk Irish culture, purple Guinness, long wet winter women, Conor McGregor/Paulie Malignaggi rumors, guy code, street fights, O.J. Simpson, Trump, freaky girls, HCO's and much more.

There is a lot of private, personal information stored within health care organizations. How do you manage and ensure you are covered for your data security concerns?

The gang is back to review the final episode of 2015 in a special Deluxe Dance Edition Package.  We talk a so-so mystery, our thoughts on Tory's replacement and we give HCO's on the Rick/Kate discussion that is way over due.

In a special deluxe dance package episode discussion the gang is back to talk the final episode of 2015.  They share their thoughts on Vikram (we hate him), Pearlmutter and our HCO's on the long overdue Kate & Rick discussion.

Hear from RoRo Eustache, Board members, Leadership & short term workers tell about the work and impact of HCO on lives.

Background: Cholangiocytes expose a striking resistance against bile acids: while other cell types, such as hepatocytes, are susceptible to bile acid-induced toxicity and apoptosis already at micromolar concentrations, cholangiocytes are continuously exposed to millimolar concentrations as present in bile. We present a hypothesis suggesting that biliary secretion of HCO(3)(-) in man serves to protect cholangiocytes against bile acid-induced damage by fostering the deprotonation of apolar bile acids to more polar bile salts. Here, we tested if bile acid-induced toxicity is pH-dependent and if anion exchanger 2 (AE2) protects against bile acid-induced damage. Methods: A human cholangiocyte cell line was exposed to chenodeoxycholate (CDC), or its glycine conjugate, from 0.5 mM to 2.0 mM at pH 7.4, 7.1, 6.7 or 6.4, or after knockdown of AE2. Cell viability and apoptosis were determined by WST and caspase-3/-7 assays, respectively. Results: Glycochenodeoxycholate (GCDC) uptake in cholangiocytes is pH-dependent. Furthermore, CDC and GCDC (pK(a) 4-5) induce cholangiocyte toxicity in a pH-dependent manner: 0.5 mM CDC and 1 mM GCDC at pH 7.4 had no effect on cell viability, but at pH 6.4 decreased viability by >80% and increased caspase activity almost 10- and 30-fold, respectively. Acidification alone had no effect. AE2 knockdown led to 3- and 2-fold enhanced apoptosis induced by 0.75 mM CDC or 2 mM GCDC at pH 7.4. Discussion: These data support our hypothesis of a biliary HCO(3)(-) umbrella serving to protect human cholangiocytes against bile acid-induced injury. AE2 is a key contributor to this protective mechanism. The development and progression of cholangiopathies, such as primary biliary cirrhosis, may be a consequence of genetic and acquired functional defects of genes involved in maintaining the biliary HCO(3)(-) umbrella. Copyright (C) 2011 S. Karger AG, Basel

Background/Aims: Ursodeoxycholic acid ( UDCA) decreases biliary secretion of cholesterol and is therefore used for the dissolution of cholesterol gallstones. It remains unclear whether these changes in biliary cholesterol excretion are associated with changes in cholesterol synthesis and bile acid synthesis. We therefore studied the activities of rate-limiting enzymes of cholesterol synthesis and bile acid synthesis, 3-hydroxy-3-methyl-glutarylcoenzyme A reductase and cholesterol 7alpha-hydroxylase, respectively, in normal subjects during UDCA feeding. Methods: UDCA was given to 8 healthy volunteers ( 5 men, 3 women; age 24-44 years) in a single dose of 10-15 mg/kg body weight for 40 days. Before and during ( days 3, 5, 10, 20, 30 and 40) UDCA treatment, urinary excretion of mevalonic acid and serum concentrations of 7alpha-hydroxy-4-cholesten-3-one (7alpha-HCO) were determined as markers of cholesterol and bile acid synthesis, respectively. The Wilcoxon signed rank test and Spearman's rank correlation coefficient were used for statistical analysis. Results: Cholesterol synthesis and serum lipid concentrations remained unchanged during UDCA treatment for 40 days. However, synthesis of bile acids increased during long-term treatment with UDCA as reflected by an increase in 7alpha-HCO serum concentrations from 39.7 +/- 21.3 ng/ml (median 32.8 ng/ml) before treatment to 64.0 +/- 30.4 ng/ml (median 77.5 ng/ml) at days 30-40 of UDCA treatment ( p < 0.05). Conclusions: UDCA treatment does not affect cholesterol synthesis in the liver, but does increase bile acid synthesis after prolonged treatment. This may represent a compensatory change following decreased absorption of endogenous bile acids as observed with UDCA therapy.