Podcasts about lacz

Már aláírt egy ügyvédi irodához Laczó Adrienn, aki lemondott bírói tisztségéről Telex     2025-01-07 11:43:22     Belföld Az egyik legnagyobb, gazdasági bűncselekményekkel foglalkozó budapesti ügyvédi irodánál fog dolgozni. Biztosítást kell kötniük az építőiparosoknak 24.hu     2025-01-07 16:27:57     Gazdaság Építőipar A legkisebb vállalkozásokra is vonatkozik a kötelezettség, azokra is, akik csak javítanak és alig van bevételük. Magyar Péter is reagált a hírre, hogy visszavonul Georg Spöttle Media1     2025-01-07 13:56:31     Karrier Magyar Péter Komment Georg Spöttle A politikus az egyik kormányközelbe került hírportál Facebook-oldalán kommentálta a PestiSrácok volt munkatársának visszavonulását. Olyannyira nem igaz, hogy nem jönnek uniós pénzek, hogy már 4 ezer milliárdot elköltött a kormány vg.hu     2025-01-07 14:40:08     Belföld Valamiért a magyar nyilvánosságban az terjedt el, hogy Magyarországra nem érkeznek az uniós források, csakhogy ez a számok alapján biztosan nem igaz. Minden negyedik magyar javítaná emésztése működésén, de nem tudja, hogyan Helló Sajtó!     2025-01-07 09:53:24     Belföld Az új év kezdete sokak számára az egészségükre való tudatosabb odafigyelés időszaka. Egy friss hazai kutatás szerint azonban a magyarok jelentős része tanácstalan, ha emésztési jóllétük javításáról van szó, annak ellenére, hogy minden negyedik ember szeretne ezen változtatni. Száguldozó BMW-sekkel indult 2025 Bács-kiskun vármegyében Autónavigátor     2025-01-07 15:11:11     Autó-motor Rendőrség BMW Tompa Már január első napján rögzített a rendőrség jelentős sebességtúllépést Bács-Kiskun vármegyében, a konkrét esetben Tompa belterületén. A közzétett fotó alapján Kínai, nálunk gyárat épít, kint feketelistára került. Mi az? Forbes     2025-01-07 10:15:05     Külföld USA Kína CATL A CATL szerint tévesen kerültek fel az amerikai védelmi minisztérium feketelistájára. Mit jelent ez valójában? Zelenszkij nem csak Orbánt, Lukasenkót is bele akarja rángatni a háborúba 444.hu     2025-01-07 14:31:03     Külföld Ukrajna háború Volodimir Zelenszkij Fehéroroszország Belarusz Alekszandr Lukasenko Legalábbis a belarusz elnök szerint, aki úgy véli: a belarusz diktatúra jobb az ukrán demokráciánál. Gyereket nevel? Így kerülheti el, hogy idén kétszer nyilatkozzon Azenpenzem     2025-01-07 10:06:00     Gazdaság Adókedvezmény Családi adókedvezmény A családi adókedvezmény nyáron emelkedik, így akik az adóelőleg-nyilatkozatban az összeget tüntetik fel, azoknak júliustól újra kell kérniük a lehetőséget. Ezt elkerülhetik, ha most januárban a gyerekek számát adják meg. Elon Musk szerint Európa hamarosan eltűnik Magyar Hírlap     2025-01-07 13:36:00     Külföld Elon Musk A The New York Times szerint a milliárdos komolyan aggódik a születési ráta csökkenése miatt. Már most óriási bajt okozott a gyengülő forint Privátbankár     2025-01-07 10:15:03     Gazdaság Az egy évvel korábbiakat 7,9, az előző havit 4,3 százalékkal múlták felül az ipari termelői árak tavaly novemberben. Botka Endre: Az Anderlecht elleni mérkőzés volt a töréspont Demokrata     2025-01-07 11:35:00     Foci A kispesti nevelésű játékos lila-fehérre cserélte a zöld-fehér mezét. Energikus focit ígér és 25 évnyi játék után sokat követel majd a futballistáitól a Fradi új edzője Telex     2025-01-07 14:57:31     Foci Magyar foci FTC Tottenham Hotspur Robbie Keane az ír válogatott és a Tottenham Hotspur csatáraként legenda lett, meg nem alkuvó hozzáállása és képességei miatt a szurkolók kedvencévé vált. Vezetőedzőként még nem sokat bizonyított, de a játékosként szerzett tapasztalatai és morálja sokat adhat a Ferencvárosnak is. Egymást váltják a frontok Kiderül     2025-01-07 12:12:34     Időjárás Orvosmeteo Ahogy áthaladt a keddi hidegfront már láthatáron a következő melegfront szerdára, és hidegfront péntekre virradóan. Bár átmenetileg pár fokkal csökkent a hőmérséklet, a megélénkülő délies szél ismét enyhébb levegőt szállít, míg péntektől vissza nem tér a tél. A további adásainkat keresd a podcast.hirstart.hu oldalunkon.

Már aláírt egy ügyvédi irodához Laczó Adrienn, aki lemondott bírói tisztségéről Telex     2025-01-07 11:43:22     Belföld Az egyik legnagyobb, gazdasági bűncselekményekkel foglalkozó budapesti ügyvédi irodánál fog dolgozni. Biztosítást kell kötniük az építőiparosoknak 24.hu     2025-01-07 16:27:57     Gazdaság Építőipar A legkisebb vállalkozásokra is vonatkozik a kötelezettség, azokra is, akik csak javítanak és alig van bevételük. Magyar Péter is reagált a hírre, hogy visszavonul Georg Spöttle Media1     2025-01-07 13:56:31     Karrier Magyar Péter Komment Georg Spöttle A politikus az egyik kormányközelbe került hírportál Facebook-oldalán kommentálta a PestiSrácok volt munkatársának visszavonulását. Olyannyira nem igaz, hogy nem jönnek uniós pénzek, hogy már 4 ezer milliárdot elköltött a kormány vg.hu     2025-01-07 14:40:08     Belföld Valamiért a magyar nyilvánosságban az terjedt el, hogy Magyarországra nem érkeznek az uniós források, csakhogy ez a számok alapján biztosan nem igaz. Minden negyedik magyar javítaná emésztése működésén, de nem tudja, hogyan Helló Sajtó!     2025-01-07 09:53:24     Belföld Az új év kezdete sokak számára az egészségükre való tudatosabb odafigyelés időszaka. Egy friss hazai kutatás szerint azonban a magyarok jelentős része tanácstalan, ha emésztési jóllétük javításáról van szó, annak ellenére, hogy minden negyedik ember szeretne ezen változtatni. Száguldozó BMW-sekkel indult 2025 Bács-kiskun vármegyében Autónavigátor     2025-01-07 15:11:11     Autó-motor Rendőrség BMW Tompa Már január első napján rögzített a rendőrség jelentős sebességtúllépést Bács-Kiskun vármegyében, a konkrét esetben Tompa belterületén. A közzétett fotó alapján Kínai, nálunk gyárat épít, kint feketelistára került. Mi az? Forbes     2025-01-07 10:15:05     Külföld USA Kína CATL A CATL szerint tévesen kerültek fel az amerikai védelmi minisztérium feketelistájára. Mit jelent ez valójában? Zelenszkij nem csak Orbánt, Lukasenkót is bele akarja rángatni a háborúba 444.hu     2025-01-07 14:31:03     Külföld Ukrajna háború Volodimir Zelenszkij Fehéroroszország Belarusz Alekszandr Lukasenko Legalábbis a belarusz elnök szerint, aki úgy véli: a belarusz diktatúra jobb az ukrán demokráciánál. Gyereket nevel? Így kerülheti el, hogy idén kétszer nyilatkozzon Azenpenzem     2025-01-07 10:06:00     Gazdaság Adókedvezmény Családi adókedvezmény A családi adókedvezmény nyáron emelkedik, így akik az adóelőleg-nyilatkozatban az összeget tüntetik fel, azoknak júliustól újra kell kérniük a lehetőséget. Ezt elkerülhetik, ha most januárban a gyerekek számát adják meg. Elon Musk szerint Európa hamarosan eltűnik Magyar Hírlap     2025-01-07 13:36:00     Külföld Elon Musk A The New York Times szerint a milliárdos komolyan aggódik a születési ráta csökkenése miatt. Már most óriási bajt okozott a gyengülő forint Privátbankár     2025-01-07 10:15:03     Gazdaság Az egy évvel korábbiakat 7,9, az előző havit 4,3 százalékkal múlták felül az ipari termelői árak tavaly novemberben. Botka Endre: Az Anderlecht elleni mérkőzés volt a töréspont Demokrata     2025-01-07 11:35:00     Foci A kispesti nevelésű játékos lila-fehérre cserélte a zöld-fehér mezét. Energikus focit ígér és 25 évnyi játék után sokat követel majd a futballistáitól a Fradi új edzője Telex     2025-01-07 14:57:31     Foci Magyar foci FTC Tottenham Hotspur Robbie Keane az ír válogatott és a Tottenham Hotspur csatáraként legenda lett, meg nem alkuvó hozzáállása és képességei miatt a szurkolók kedvencévé vált. Vezetőedzőként még nem sokat bizonyított, de a játékosként szerzett tapasztalatai és morálja sokat adhat a Ferencvárosnak is. Egymást váltják a frontok Kiderül     2025-01-07 12:12:34     Időjárás Orvosmeteo Ahogy áthaladt a keddi hidegfront már láthatáron a következő melegfront szerdára, és hidegfront péntekre virradóan. Bár átmenetileg pár fokkal csökkent a hőmérséklet, a megélénkülő délies szél ismét enyhébb levegőt szállít, míg péntektől vissza nem tér a tél. A további adásainkat keresd a podcast.hirstart.hu oldalunkon.

Join Andy this week as he sits down with Susan Lacz, the CEO of Ridgewells Catering. With a career spanning over three decades, Susan has been a pivotal force in the catering and events industry, steering Ridgewells Catering through significant transformations and challenges. Listen in as Susan shares her inspiring journey, starting with her early influences, notably her grandmother, whose culinary skills and hospitality left a lasting impression on her. She discusses her initial foray into the food business, from working at a deli in Wyckoff, New Jersey, to bartending in Washington, D.C. Her passion for sales and marketing, paired together with her love for food, set the stage for her ascent in the catering world. In this candid conversation, Susan recounts the challenges and triumphs of taking over Ridgewells Catering in 1997, a company with deep roots dating back to the 1920s. She describes her mission to restore the company's former glory by emphasizing quality, creativity, and fresh culinary approaches. She reflects on key milestones, such as firing her former boss to initiate positive changes and hosting an innovative rebranding event at the Halcyon House. Susan emphasizes the importance of culture in her company, highlighting the sense of ownership and appreciation that permeates the workplace. She and Andy also touch upon the broader shifts in the catering and events industry, from the increasing focus on decor and presentation to the critical role of staying ahead of trends. Susan's insights into navigating the saturated market, her strategic decision to divest from rental equipment, and her recent exclusive venue management agreements offer valuable lessons for industry professionals, and Andy hopes that you enjoy listening to this conversation with her! If so, then please don't keep it to yourself! He would appreciate it if you were to share it with a few good friends and industry colleagues who might also enjoy it! Andy would also appreciate it if you were to leave a top review wherever you listen to the show because that really helps the show out! Lastly, don't forget to follow The Wedding Biz on Instagram and subscribe to the podcast so that you never miss a new episode when it drops!   Have you heard about Stop and Smell the Roses with Preston Bailey on The Wedding Biz Network? Listen as Preston shares the secrets, tools, and technologies behind his extraordinary ability to create a theatrical environment out of any space. Also, don't forget about Sean Low's podcast The Business of Being Creative, where Sean discusses the power of being niched, pricing strategies, metrics of success, and so much more. You can find both shows on The Wedding Biz Network.   SUPPORTING THE WEDDING BIZ Become a patron and support Andy and the show! If you are so inspired, contribute!   Time Stamps [1:01] - Today's guest is Susan Lacz, CEO of Ridgewells Catering. [1:57] - Susan's grandmother, an immigrant from Poland, was her strong role model and inspiration in cooking. [3:16] - Susan explains that she is a salesperson first and enjoyed selling food and beverages, including pastrami sandwiches and drinks. [4:18] - Susan wanted to restore the company's original quality and allure after multiple ownership changes. [5:19] - Hear about some of Susan's milestones, such as firing her boss, focusing on quality, hiring new culinary leaders, and creatively rebranding Ridgewell. [6:54] - Susan prioritized creating a strong, appreciative culture, guided by mentors including her entrepreneurial father. [9:34] - Hear about some of the challenges Susan faced, especially during COVID, and how she credits her team. [11:25] - Susan noted the shift from food to decor in the industry and evolved by focusing on culinary excellence. [13:35] - Susan is excited about managing the iconic Mellon Auditorium, now a preferred caterer there, which opens new opportunities for her team. [14:53] - Prioritizing balance and mentorship, Susan believes in taking care of herself to be a good mother and leader. [16:26] - How does Susan define success? [17:15] - Andy reveals where Susan can be followed online. LINKS AND RESOURCES Videos: Ridgewells Catering - “Erin Go Glam 2024”   Find Susan: Susan's LinkedIn Page Ridgewells Catering - Website Ridgewells Catering - Instagram Page Ridgewells Catering - Facebook Page   Follow The Wedding Biz on Social: The Wedding Biz The Wedding Biz on Instagram: @theweddingbiz The Wedding Biz on Facebook: @theweddingbiz The Wedding Biz Network The Music Makers   Support The Wedding Biz by clicking here.   Title Sponsor: This episode is sponsored by Kushner Entertainment.  

Hello, my name is Feren Laczo, I am an editor at the Review of Democracy, and I am also the co-head of the Ideas section. And it is my pleasure today to share with you a brief list of some of the most impressive publications we have covered this year. Ideas editors and podcasters have been invited to a continuous feast in 2023: the year has offered an unusual number of original publications of the highest caliber. Natasha Wheatley's The Life and Death of States: Central Europe and the Transformation of Modern Sovereignty, the Vienna-based discussion of which we were proud to co-organize; Sam Moyn's Liberalism Against Itself. Cold War Intellectuals and the Making of Our Times that substantially critiques the dominant form Western liberalism has taken; Danielle Allen's exciting proposal of a power-sharing liberalism; George Steinmetz's major monograph on the colonial origins of modern social thought in France; or Adam Shatz's collection of essays on the radical imagination have all been evident highlights. Here comes an all too selective list of five recommendations from RevDem Ideas of books that deserve to be more widely read and discussed.

Gillian Coughlan is a postdoc whose work focuses on the role of spatial navigation in dementia. In this conversation, we talk about how Gillian went from Ireland to doing a PhD in the UK, different ways for diagnosing Alzheimer's disease, what beta-amyloid and tau are, what spatial navigation has to do with dementia, and whether early menopause can affect women's spatial navigation performance and risk of getting dementia.BJKS Podcast is a podcast about neuroscience, psychology, and anything vaguely related, hosted by Benjamin James Kuper-Smith.Support the show: https://geni.us/bjks-patreonTimestamps00:00: Playing the piano07:13: How Gillian ended up doing her PhD with Michael Hornberger in Norwich14:02: How to find a good mentor16:48: Sea Hero Quest22:28: Diagnosing Alzheimer's disease32:37: The role of Beta-Amyloid and tau in dementia34:41: Spatial navigation, the entorhinal cortex, and dementia44:14: Does menopause affect spatial navigation and risk of dementia?50:31: Book or paper more people should read52:37: Something Gillain wishes she'd learnt sooner55:31: Advice for PhD students and postdocsPodcast linksWebsite: https://geni.us/bjks-podTwitter: https://geni.us/bjks-pod-twtGillian's linksGoogle Scholar: https://geni.us/coughlan-scholarTwitter: https://geni.us/coughlan-twtBen's linksWebsite: https://geni.us/bjks-webGoogle Scholar: https://geni.us/bjks-scholarTwitter: https://geni.us/bjks-twtReferences and linksEpisodes with Michael Hornberger and Hugo Spiershttps://geni.us/bjks-hornbergerhttps://geni.us/bjks-spiers  Coughlan, DeSouza, Zhukovsky, Hornberger, Grady & Buckley (2023). Spatial cognition is associated with levels of phosphorylated-tau and β-amyloid in clinically normal older adults. Neurobiology of Aging.Coughlan, ... Buckley (2023). Association of age at menopause and hormone therapy use with tau and β-amyloid positron emission tomography. JAMA Neurology.Coughlan, Coutrot, Khondoker, Minihane, Spiers & Hornberger (2019). Toward personalized cognitive diagnostics of at-genetic-risk Alzheimer's disease. PNAS.Coughlan, Laczó, Hort, Minihane & Hornberger (2018). Spatial navigation deficits—overlooked cognitive marker for preclinical Alzheimer disease?. Nature Reviews Neurology.Eger (2017). The Choice.Pertesi, Coughlan, Puthusseryppady, Morris & Hornberger (2019). Menopause, cognition and dementia–A review. Post reproductive health.

Overlapping crises, enforced political passivity and a new political normal: all things that gradually dismantle a democracy. Long standing Eurozine contributor, historian Ferenc Laczó joins editor-in-chief Réka Kinga Papp to discuss how a democracy can be alive and dead at the same time. Laczó took part in the discussion about how democracies die in the Eurozine focal point ‘The writing on the wall' with his article 'How democracies transform, fast and slow' : https://www.eurozine.com/how-democracies-transform-fast-and-slow/ A longer version of this conversation with bonus material is available to Eurozine's patrons. Support our work with as little as €5/month or more to access extra content and help sustain this publication that connects you with over 100 publications from across Europe. www.patreon.com/Eurozine Please subscribe to the podcast and leave a review so more people can find us. You can also subscribe to our weekly newsletter, so you'll always know what's worth thinking about: www.eurozine.com/newsletter/

United States Navy SEAL Veteran Kevin Lacz joins Sid to tell his Memorial Day story and what being a Navy SEAL veteran means to him. Lacz spoke on his experience alongside Chris Kyle as an American sniper and what it really takes to put everything on the line for our country. Learn more about your ad choices. Visit megaphone.fm/adchoices

V dnešní epizodě k nám hovořil pan prof. MUDr. Jan Laczó, Ph.D, který působí jako lékař v Kognitivním centru ve Fakultní nemocnici Motol. Zaobírá se diagnostikou a léčbou onemocnění, které způsobují postižení vyšších funkcí mozku, především Alzheimerovou nemocí. Témata byly: Je častější Alzheimerova choroba pro ženy nebo muže? A proč? Jaké jsou první příznaky, kterých si můžou blízcí všimnout? Jaké jsou projevy Alzheimerovy nemoci? Jaká je prevence?

In this episode of Partnering Leadership, Susan Lacz, principal, and chief executive officer of Ridgewells Catering, shared her leadership journey with Mahan Tavakoli. Susan Lacz spoke about how she has managed the obstacles in her life and has built a successful business that needs to continue changing and adapting through disruption. Susan Lacz also shared why she is so committed to giving back to the community through Ridgwell Catering's support and giving her time and energy to many institutions and organizations, including Marymount University and Junior Achievement of Greater Washington.Some Highlights:-How Susan Lacz's Polish culture influenced her love of entertaining-Susan Lacz on overcoming a significant setback early in life-Rebranding strategies contributing to Ridgewells Catering's continued growth and expansion-Growth opportunities through Young President's Organization.  -How Susan Lacz led her team through the most brutal stretch of the pandemic.  -Susan Lacz embracing an antifragile mindset in leadership and life-Why mentorship is essential to becoming more impactful leadersAlso mentioned in this episode:-Linda Rabbit, founder and Chairman of Rand Construction Corporation (Listen to Linda's episode on Partnering Leadership here)-Marymount University (Listen to Marymount's President Dr. Irma Bacerra on Partnering Leadership here)Connect with Susan Lacz :Ridgewells CateringSusan Lacz on LinkedinConnect with Mahan Tavakoli:MahanTavakoli.comMore information and resources available at the Partnering Leadership Podcast website: PartneringLeadership.com

In this conversation with Bence Bari and Orsolya Sudár, editors Ferenc Laczó and Bálint Varga and contributor Dóra Vargha discuss the new volume Magyarország globális története, 1869-2022 (A Global History of Hungary, 1869-2022). The conversation focuses on some of the innovative questions posed by trying to reconceptualize the history of a Central and Eastern European country in a global frame; how the subjects of the volume's one hundred chapters have been selected; the relation of this new book to other narratives of Hungarian history; and the more political stakes of releasing such a publication today. László Bence Bari is a doctoral candidate at the History Department of the Central European University. Ferenc Laczó is an assistant professor with tenure (universitair docent 1) in history at Maastricht University. He is also an editor at the Review of Democracy. Orsolya Sudár is a doctoral candidate at the History Department of the Central European University. Bálint Varga is a research fellow at the Center for Humanities in Budapest. Dóra Vargha is professor of history and medical humanities at the University of Exeter and Humboldt University, and is a visiting scholar at the Max Planck Institute for the History of Science, Berlin.

A választás utáni levegővételnek vége: miniszterek és államtitkárok kinevezve, kezdődik az új ciklus. Az ötödik Orbán-kormány első komolyabb intézkedése a honvédelmi és a rezsialap létrehozása, a kabinet különböző extraprofitot felhalmozó szektorok megadóztatásával igyekszik saját szavaival élve megvédeni a családokat. Mit jelent ez valóságban? Elég a gazdaság rendberázásához az a 800 milliárdos összeg, amit az alapoktól remélnek? Nem rakódhatnak rá a terhek a fogyasztókra? Elemzőinkkel Pogátsa Zoltánnal és Zsiday Viktorral beszéljük át a kormányinfón elhangzottak összefüggéseit. De nem feledkezünk meg a kultúráról sem: Szabó László, a Magyar Teátrumi Társaság titkára meghirdette a kultúrharc újabb felvonását. A Kommentár folyóiratban közlendő programadó írásáról kérdezzük stúdiónkban. Vendégünk lesz még Tóth Viktor, nemrégiben felmondott pedagógus, és bemutatjuk a Magyarország globális története című kiadványt is, a szerkesztőkkel beszélgetünk.Vendégeink:- Szabó László, titkár, Magyar Teátrumi Társaság- Laczó Ferenc, történész, Maastrichti Egyetem- Varga Bálint, adjunktus, Grazi Egyetem- Tóth Viktor, pedagógus- Zsiday Viktor, közgazdász- Pogátsa Zoltán, közgazdász★ Support this podcast on Patreon ★

This week, please join author Vasan Ramachandran and Associate Editor Mercedes Carnethon as they discuss the article "Temporal Trends in the Remaining Lifetime Risk of Cardiovascular Disease Among Middle-Aged Adults Across 6 Decades: The Framingham Study." Dr. Carolyn Lam: Welcome to Circulation on the Run, your weekly podcast, summary and backstage pass to the journal and its editors. We're your co-hosts. I'm Dr. Carolyn Lam, Associate Editor from the National Heart Center in Duke National University of Singapore. Dr. Greg Hundley: And I'm Dr. Greg Hundley, Associate Editor, Director of the Pauley Heart Center at VCU Health in Richmond, Virginia. Dr. Carolyn Lam: Greg, I'm so excited about today's feature paper. You see, I trained at the Framingham Heart Study and today's feature paper talks about the temporal trends in the remaining lifetime risk of cardiovascular disease among middle aged adults across six decades in the Framingham Heart Study. Truly a landmark study and a discussion nobody wants to miss. But first, let's talk about the other papers in today's issue, and I understand that you've got one ready. Dr. Greg Hundley: You bet Carolyn. I'll get started first. Thank you. So my first paper comes from Dr. Daniel Mark from Duke University and it refers to the ISCHEMIA trial. Dr. Carolyn Lam: Ooh, could you please first remind us what is the ISCHEMIA trial and are you presenting a substudy, is that correct? Dr. Greg Hundley: Right, Carolyn. So the International Study of Comparative Health Effectiveness with Medical and Invasive Approaches, or ISCHEMIA, compared an initial invasive strategy with an initial conservative strategy in 5,179 participants with chronic coronary disease and moderate or severe ischemia. And this sub study of the ischemia research program included a comprehensive quality of life analysis. Dr. Carolyn Lam: So very interesting. What did they find Greg? Dr. Greg Hundley: Right, Carolyn. So this study included 1,819 participants. 907 in the invasive, 912 in the conservative. And collected a battery of disease specific and generic quality of life instruments by structured interviews at baseline. And then at three, 12, 24 and 36 months post randomization, and then finally at study closeout. Now Carolyn, these assessments included an angina related quality of life assessment from the 19 item Seattle Angina Questionnaire, a generic health status assessment, an assessment of depressive symptoms, and for North American patients, cardiac functional status from the Duke Activity Status Index, or DASI. In this study, Carolyn, in terms of results, the median age was 67 years and about 20% were women and about 16% were nonwhite. So Carolyn, getting to the results. The estimated mean difference for the SAQ 19 summary score favored invasive therapy. And remember the SAQ 19 was the Seattle Angina Questionnaire. Dr. Greg Hundley: Next, no differences were observed in patients with rare or absent baseline angina. Next, among patients with more frequent angina baseline, those randomized to invasive had a mean point higher score on the SAQ 19 summary score than the conservative approach, with consistent effects across all of the SAQ subscales including physical limitations, angina frequency and quality of life health perceptions. For the DASI, and remember DASI refers to the Duke Activity Status Index, no difference was estimated overall by treatment. But in patients with baseline marked angina, DASI scores were higher for the interventional arm. Whereas patients with rare or absent baseline angina showed really no treatment related differences. Dr. Carolyn Lam: Oh, okay. So a lot of results. What's the take-home message, Greg? Dr. Greg Hundley: Right, Carolyn. Glad you asked. So in the ISCHEMIA comprehensive quality of life substudy, patients with more frequent baseline angina reported greater improvements in the symptom physical functioning and psychological wellbeing dimensions of quality of life when treated with an invasive strategy. Whereas patients who had rare or absent angina baseline reported no consistent treatment related quality of life differences. Dr. Carolyn Lam: Wow. Thank you, Greg. Very interesting indeed. Now from angina to now cholesterol. Now, cholesterol guidelines typically prioritize primary prevention statin therapy based on 10 year risk of cardiovascular disease. Now the advent of generic pricing may in fact justify expansion of statin eligibility. Moreover, 10 year risk may not be the optimal approach for statin prioritization. So these issues were looked at in this next paper by authors led by Dr. Kohli Lynch from Northwestern University and colleagues who estimated the cost effectiveness of expanding preventive statin eligibility, and evaluated novel approaches to prioritization from a Scottish health sector perspective. A computer simulation model predicted long term health and cost outcomes in Scottish adults, age 40 years or more. Dr. Greg Hundley: So Carolyn, what did they find? Dr. Carolyn Lam: The advent of generic pricing has rendered preventive statin therapy cost effective for many adults. Absolute risk reduction guided statin therapy, which is based on 10 year cardiovascular disease risk and non HDL cholesterol levels, is cost effective and would improve population health. Whereas age stratified risk thresholds were more expensive and less effective than alternative approaches to statin prioritization. So guidelines committees may need to expand statin eligibility and consider new ways to allocate statins based on absolute risk reduction rather than 10 year risk thresholds. Dr. Greg Hundley: Very nice Carolyn. Always important, new information regarding statin therapy. Well Carolyn, my next paper comes to us from the world of preclinical science. And Carolyn, as you know, the regenerative capacity of the heart after myocardial infarction is limited. And these authors led by Dr. Tamer Mohamed from University of Louisville previously showed that ectopic introduction of Cdk1, CyclinB1 and Cdk4, CyclinD1 complexes and we'll refer to those now as 4F, promoted cardiomyocyte proliferation in 15 to 20% of infected cardiomyocytes in vitro and in vivo and improved cardiac function after MI in mice. So Carolyn, in this study using temporal single cell RNA sequencing, the investigative team aimed to identify the necessary reprogramming stages during the forced cardiomyocyte proliferation with 4F on a single cell basis. And also using rat and pig models of ischemic heart failure, they aim to start the first preclinical testing to introduce 4F gene therapy as a candidate for the treatment of ischemia induced heart failure. Dr. Carolyn Lam: Oh, wow Greg. So what did they find? Dr. Greg Hundley: Several things, Carolyn. First, temporal bulk and single cell RNA sequencing and further biochemical validations of mature HIPS cardiomyocytes treated with either LAcZ or 4F adenoviruses revealed full cell cycle reprogramming in 15% of the cardiomyocyte population at 48 hours post-infection with 4F. Which was mainly associated with sarcomere disassembly and metabolic reprogramming. Second Carolyn, transient overexpression of 4F specifically in cardiomyocytes was achieved using a polycistronic non-integrating lentivirus encoding the 4F with each driven by a TNNT2 promoter entitled TNNT2-4F polycistronic-NIL. Now this TNNT2-4F polycistronic-NIL or control virus was injected intra myocardial one week after MI in rats, so 10 per group, and pigs, six to seven per group. Dr. Greg Hundley: And four weeks post-injection the TNNT2-4F polycistronic-NIL treated animals showed significant improvement in left ventricular injection fraction and scar size compared with the control virus treated animals. And four months after treatment, the rats that received TNNT2-4F polycistronic-NIL still showed a sustained improvement in cardiac function without obvious development of cardiac arrhythmias or systemic tumorigenesis. And so Carolyn this study advances concepts related to myocellular regeneration by providing mechanistic insights into the process of forced cardiomyocyte proliferation and advances the clinical feasibility of this approach by minimizing the oncogenic potential of the cell factors, thanks to the use of a novel transient and cardiomyocyte specific viral construct. Dr. Carolyn Lam: Wow. What a rich study. Thanks so much, Greg. Dr. Greg Hundley: Well, Carolyn, how about if we see what else and what other articles are in this issue. And maybe I'll go first. So there's a research letter from Dr. Wu entitled Modeling Effects of Immunosuppressive Drugs on Human Hearts Using IPSC Derived Cardiac Organoids and Single Cell RNA Sequencing. Carolyn, there's an EKG challenge from Dr. Yarmohammadi, entitled “Fast and Furious, A Case of Group Beating in Cardiomyopathy.” And then finally from Dr. Tulloch, a really nice Perspective entitled “The Social Robots are Coming, Preparing For a New Wave of Virtual Care in Cardiovascular Medicine. Dr. Carolyn Lam: Oh, how interesting. Well, also in the mail back is an exchange of letters of among Drs. Lakkireddy, Dhruva, Natale, and Price regarding Amplatzer Amulet Left Atrial Appendage Occluder versus Watchman Device for stroke prophylaxis, a randomized control trial. All right. Thank you so much, Greg. Shall we go on to our feature discussion now? Dr. Greg Hundley: You bey. Welcome listeners to our feature discussion today. And we're so fortunate we have with us today, Dr. Vasan Ramachandran from Boston University and our own Associate Editor, Dr. Mercedes Carnethon from Northwestern University in Chicago. Welcome to you both. And Vasan, let's start with you. Could you describe for us some of the background information pertaining to your study and what was the hypothesis that you wanted to address? Dr. Vasan Ramachandran: Thank you, Greg, first of all for having me. So we know two facts. One is that heart disease and stroke disease death rates and incidents are declining over the last six decades in the United States. Juxtapose against that is also the observation that there is rising incidence of obesity and overweight, and also a rising burden of diabetes. There are a lot of advances in our ability to treat high blood pressure, high cholesterol, as well as high blood sugar. So we wanted to ask the question, given the historic trends in control awareness of risk factors and their control, interrupted by this escalating burden of obesity, overweight, and diabetes, what is the lived experiences of people over time in terms of the risk of developing heart disease or stroke using a metric we call as the remaining lifetime risk of developing heart disease or stroke. Dr. Greg Hundley: The hypothesis you wanted to address? Dr. Vasan Ramachandran: The hypothesis we wanted to address was that perhaps the decline in the incidence of heart disease and stroke may have decreased over time given the escalating burden of overweight, obesity and diabetes. Dr. Greg Hundley: Very nice. And can you describe for us your study population and your study design? Dr. Vasan Ramachandran: Thank you, Greg. So the Framingham Heart Study is one of the oldest running epidemiological studies in the world. We have multiple cohorts. The study began in 1948 with the original cohort, the offspring cohort enrolled in 1971, third generation cohort in 2002, and two minoritized cohorts in the 1990s and 2002. So we have an observation period of different cohorts over a six decade period. So we asked the question, if you were a participant in the Framingham study between 1960 and 1979 and then 1980 to 1999, and then 2000 to 2018, what was your lifetime risk of experiencing a heart disease or stroke in the three different time periods? Is it going down, is it steady or is it going up? Dr. Greg Hundley: Very nice. And so, Vasan, describe your study results. Dr. Vasan Ramachandran: Look, what we found was if you look at the first, the 20 year period from 1960 to 1979, and compare that with the latest, which is 2000 to 2018, in the initial time period, the lifetime risk of developing heart disease or stroke in a man was pretty high. It was about one in two. And that for a woman was about one in three. So when you come to the latest epoch, what we find that the risk of one in two men had dropped to about one in three men in the latest decade. For women, the risk declined from what was one in three in the earlier epoch to one in four. So approximately there was about a 36% reduction in the lifetime probability of developing heart disease or stroke across the six decade period of observation. Dr. Greg Hundley: Very nice. And so help us a little bit, put the context of your results into what that might mean for us today as we are managing patients with atherosclerotic disease. Dr. Vasan Ramachandran: Yes, Greg. What it means is that the permeation of the advances in science in terms of the screening of risk factors, awareness of risk factors, medications to lower these risk factors effectively, the clinical trials that have given us these new medications, they may have translated into a reduction in risk over time. That the lived experience of people in the later decades is better in terms of having a lower risk of heart disease or stroke as the consequence of multiple advances that have happened in heart disease and stroke. Dr. Greg Hundley: Well, thank you so much Vasan. Well listeners, now we're going to turn to our Associate Editor, Mercy Carnethon. And Mercy, you have many papers come across your desk. What attracted you to this particular paper and how do you put these results really in the context of other science pertaining to risk associated with populations that may have atherosclerotic cardiovascular disease? Dr. Mercedes Carnethon: Thanks so much for that question, Greg. And again, Vasan, I really thank you and your team for bringing forth such outstanding research. You know, as cardiovascular disease epidemiologists, we were all raised and taught that what we know about risk factors for cardiovascular disease are based on the Framingham cohort. And so I was really excited to see this very comprehensive piece of work that characterized what the Framingham study has identified and also leverages the unique characteristics of a study that started in 1948. Dr. Mercedes Carnethon: So, you know, we're almost 75 years in and actually has the ability that cross sectional studies don't have to look over longer periods of time at risk. And you know, when we think about papers that excite us, that we really want to feature in circulation, they are papers that teach us something new. And I will say there were aspects of this work that confirmed what I had heard but had not seen using empirical data. Namely that the remaining lifetime risks for developing cardiovascular disease were going down over time, and they were going down secondary to better management and recognition of the risk factors that the Framingham cohort study had really been instrumental in identifying in the first place. Dr. Mercedes Carnethon: There were surprising elements of the paper. The surprising elements being that I think as you brought up earlier, we were concerned that risk factors that were on the rise, such as obesity, were threatening these increases in life expectancy. And it was really nice to see that the findings held, even in the face of rising risk factors. And just to summarize, what I really like about this piece when we situate it within circulation, where we are addressing clinical treatment factors, where we're also featuring clinical trials and even other epidemiologic studies, is that your work identifies for us the overall context in which the clinicians who read the journal are thinking about managing patients and where we're going. It highlights our successes, but it also really brings up what we need to do next. And I look forward to hearing from you about where you think this may be headed. Dr. Greg Hundley: Well, Mercy, you're teeing us up for that next question. Vasan, what do you think is the next study or studies that need to be performed in this space? Dr. Vasan Ramachandran: Thank you, Greg and Mercy, for your kind comments. Like I shared, this is a success story for a predominantly white population in the Northeast. We are very much aware about the heterogeneity and the geographic variation in heart disease burden in our country. So one of the success stories interpretation might be this represents the upper bound. What can happen to a population that is compliant with screening of risk factors, awareness of risk factors, treatment and healthcare access. I think the next set of studies should broaden the study population to bring in additional populations that are more diverse, that are also followed up over a period of time to assess and put the current observations in the appropriate context. Do we see similar findings longitudinally in other cohorts with non-white participants? Is it different, is their lived experience different? If so, why? And that could inform us how we can reach the success story and replicate it across the entirety of our country. Dr. Greg Hundley: And Mercy, do you have anything to add? Dr. Mercedes Carnethon: I do. You know, I really like that focus on broadening to whom these results are applicable. We've undergone a lot of shifts within our country and also around the world. You know, circulation, we have a worldwide readership. I would love to see this sort of work replicated across different countries to the extent that we have the data to do so, recognizing that limitation. But I'd love to see work focus on comparing how these things change in low income countries, middle income and high income countries, so that we can really think about resource allocation and find strategies to try to replicate the successes that we are seeing based on the data from the Framingham heart and offspring studies. Dr. Greg Hundley: Excellent. Well listeners, we really appreciate the opportunity to get together today with Dr. Vasan Ramachandran from Boston University and our own Associate Editor, Dr. Mercedes Carnethon from Northwestern University in Chicago. And really appreciate them for bringing us these epidemiologic data from the Framingham cohort, indicating that over the past decades, mean life expectancy increased and the remaining lifetime risk of atherosclerotic cardiovascular disease decreased across individuals in the cohort, even after accounting for increasing incidences of other cardiovascular risk factors like obesity and smoking. Well on behalf of Carolyn and myself, we want to wish you a great week and we will catch you next week on the run. Dr. Greg Hundley This program is copyright of the American Heart Association, 2022. The opinions expressed by speakers in this podcast are their own and not necessarily those of the editors or of the American Heart Association. For more, please visit ahajournals.org.

Többet és mást is láthatunk a magyar történelemből, ha nem nemzeti fókuszból, hanem a globális trendek helyi hatásai felől közelítjük meg. Laczó Ferenc és Varga Bálint az e heti G7 Podcast vendégei. The post Mit látunk a történelemből, ha a magyar cseppen keresztül nézzük a világ tengerét? first appeared on G7 - Gazdasági sztorik érthetően.

Susan Lacz, principal and Chief Executive Officer of Ridgewells Catering,  joins me for today's episode of Notable Leaders' Radio! She's going to share powerful stories of what drives her and how that drive has built a successful and meaningful career.    She's a highly recognized leader and sought after speaker in the catering and special events industry.    When you look at Susan's impressive career,  journey and achievements, it would be easy to describe her as driven. However, Susan doesn't  agree with that. She attributes her passion for what she does and making a difference as her greatest motivator.   Tune in to hear her amazing story!   Guest Bio:   Susan Lacz, a recognized leader and sought-after speaker in the catering and special events industry, is Principal and Chief Executive Officer of Ridgewells Catering.  As an influential leader in the business community, she has been honored with the Washington Business Journal's Women Who Mean Business award and named to their Power 100 list. She has been a finalist for The Ernst & Young Entrepreneur of the Year Greater Washington Award and a recipient of the Brava! Award from SmartCEO Magazine.  Recognized for her charitable work, she was bestowed by Miriam's Kitchen with the Bowls of Compassion Award, the Greater Washington Board of Trade presented Susan and Ridgewells with its Golden Links Award, and Montgomery County's It Takes a Village Award.  Susan lives in Chevy Chase, Maryland and has two sons, Nathan and Ayden.   www.ridgewells.com   https://www.instagram.com/ridgewellsdc/    https://www.facebook.com/ridgewellsdc/   https://www.linkedin.com/in/susanlacz/  Belinda's Bio:    Belinda Pruyne is a sought-after Leadership Advisor, Coach, Consultant and Keynote speaker. She is a leading authority in guiding global executives, professionals and small business owners to become today's highly respected leaders.   She gained a wealth of expertise in the client services industry as Executive Vice President, Global Director of Creative Management at Grey Advertising, managing 500 people around the globe. With over 20+ years of leadership development experience, she brings industry-wide recognition to the executives and companies she works with. Whether a startup, turnaround, acquisition, or global corporation, executives and companies continue to turn to Pruyne for strategic and impactful solutions in a rapidly shifting economy and marketplace.    Website: Belindapruyne.com Email Address: hello@belindapruyne.com LinkedIn: https://www.linkedin.com/in/belindapruyne  Facebook: https://www.facebook.com/NotableLeadersNetwork.BelindaPruyne/  Twitter: https://twitter.com/belindapruyne?lang=en  Instagram: https://www.instagram.com/belindapruyne/ 

Michael Hornberger is a professor of applied dementia research at the University of East Anglia who developed Sea Hero Quest, a mobile game for studying spatial navigation that was downloaded more than 4 million times. In this conversation, we talk about Sea Hero Quest, how Michael (together with Hugo Spiers) developed it, the first findings, and dementia in general.BJKS Podcast is a podcast about neuroscience, psychology, and anything vaguely related, hosted by Benjamin James Kuper-Smith. New conversations every other Friday. You can find the podcast on all podcasting platforms (e.g., Spotify, Apple/Google Podcasts, etc.).Timestamps00:04: How Michael started doing research on dementia02:32: What is Sea Hero Quest?05:18: How Sea Hero Quest got started and developed31:45: Start discussing some results from Sea Hero Quest38:16: How does Sea Hero Quest relate to Michael's work on dementia?43:16: How far are we from using (something like) Sea Hero Quest as an early biomarker for Alzheimer's?45:36: What is dementia and how can we prevent it? Discussing Michael's new book about Alzheimer's 'Tangled Up'Podcast linksWebsite: https://geni.us/bjks-podTwitter: https://geni.us/bjks-pod-twtMichael's linksWebsite: https://geni.us/hornberger-webGoogle Scholar:  https://geni.us/hornberger-scholarTwitter:  https://geni.us/hornberger-twtBook: https://geni.us/hornberger-bookBen's linksWebsite: https://geni.us/bjks-webGoogle Scholar: https://geni.us/bjks-scholarTwitter:  https://geni.us/bjks-twtReferencesAlzheimer (1906). Uber einen eigenartigen, schweren Erkrankungsprozess der Hirnrinde. Neurol. Cbl..Alzheimer, Förstl, & Levy (1991). On certain peculiar diseases of old age. History of psychiatry.Anguera, Boccanfuso, Rintoul, Al-Hashimi, Faraji, Janowich, ... & Gazzaley (2013). Video game training enhances cognitive control in older adults. Nature.See also: https://en.wikipedia.org/wiki/NeuroRacerCoughlan, Coutrot, Khondoker, Minihane, Spiers & Hornberger (2019). Toward personalized cognitive diagnostics of at-genetic-risk Alzheimer's disease. Proceedings of the National Academy of Sciences.Coughlan, Laczó, Hort, Minihane, & Hornberger (2018). Spatial navigation deficits—overlooked cognitive marker for preclinical Alzheimer disease?. Nature Reviews Neurology.Coughlan, Puthusseryppady, Lowry, Gillings, Spiers, Minihane, & Hornberger (2020). Test-retest reliability of spatial navigation in adults at-risk of Alzheimer's disease. Plos one.Coutrot, Schmidt, Coutrot, Pittman, Hong, Wiener, ... Hornberger, & Spiers (2019). Virtual navigation tested on a mobile app is predictive of real-world wayfinding navigation performance. PloS one.Coutrot, Silva, Manley, de Cothi, Sami, Bohbot, ... Hornberger, & Spiers (2018). Global determinants of navigation ability. Current Biology. Fold.it game: https://fold.it/Hornberger (2021). Tangled up. The science and history of Alzheimer's disease.Kunz, Schröder, Lee, Montag, Lachmann, Sariyska, ... & Axmacher (2015). Reduced grid-cell–like representations in adults at genetic risk for Alzheimer's disease. Science.

In this episode of Partnering Leadership, Susan Lacz, principal, and chief executive officer of Ridgewells Catering, shared her leadership journey with Mahan Tavakoli. Susan Lacz spoke about how she has managed the obstacles in her life and has built a successful business that needs to continue changing and adapting through disruption. Susan Lacz also shared why she is so committed to giving back to the community through Ridgwell Catering's support and giving her time and energy to many institutions and organizations, including Marymount University and Junior Achievement of Greater Washington.Some Highlights:-How Susan Lacz's Polish culture influenced her love of entertaining-Susan Lacz on overcoming a significant setback early in life-Rebranding strategies contributing to Ridgewells Catering's continued growth and expansion-Growth opportunities through Young President's Organization.  -How Susan Lacz led her team through the most brutal stretch of the pandemic.  -Susan Lacz embracing an antifragile mindset in leadership and life-Why mentorship is essential to becoming more impactful leadersAlso mentioned in this episode:-Linda Rabbit, founder and Chairman of Rand Construction Corporation (Listen to Linda's episode on Partnering Leadership here)-Marymount University (Listen to Marymount's President Dr. Irma Bacerra on Partnering Leadership here)Connect with Susan Lacz :Ridgewells CateringSusan Lacz on LinkedinConnect with Mahan Tavakoli:MahanTavakoli.comMore information and resources available at the Partnering Leadership Podcast website: PartneringLeadership.com

Kevin Lacz był snajperem w jednej z najbardziej elitarnych jednostek na świecie. Jako członek SEAL Team Three walczy w bitwie pod Ramadi. Odsłużył dwie tury w Iraku. Widział piekło na ziemi. Mimo to twierdził, że wojna nie odcisnęła na nim trwałego piętna. Lacz opisał swoje wspomnienia w książce pod tytułem: “Mściciel. Ostatni snajper”. Tytuł słusznie sugeruje, że nie jest to raczej lektura dla ludzi o słabych nerwach. Nie mniej jednak, nawet jeśli z reguły nie czujecie się komfortowo w militarnych klimatach, to tej relacji z Iraku warto poświęcić kilka godzin. Lacz, oprócz tego, że pozwala nam zajrzeć za kulisy owianego nimbem tajemnicy szkolenia SEALs i opisuje, nie szczędząc brutalnych szczegółów, poszczególne operacje, w których uczestniczył w Iraku, to stara się również wytłumaczyć ideę, która legła u podstaw wojny z terroryzmem. W związku z premierą drugiego wydania “Mściciela” w Polsce postanowiliśmy poprosić, aby wytłumaczył nam ją osobiście. Nie zabrakło również miejsca na pytania od was - czytelników jego książki.

Was the East-West split never meant to go away, or did an uneven exchange of influences stop the European unification many had so hoped for? Some seem to forever carry the East with them, while others substitute the colloquial ‘end of history' with shallow concepts for political gain. Easterners are tired of the perpetual post-Communist stigma, while Westerners suggest the promised land wasn't quite so promising in the first place. Curators Ferenc Laczó and Luka Lisjak Gabrijelčič discuss the new Eurozine anthology. You can order the book here: https://tinyurl.com/yymveh7c Subscribe to Eurozine's newsletter: https://www.eurozine.com/newsletter/

Today I’m talking with Katie Lacz. She works for Women’s Ordination Conference, the United States branch of a worldwide movement to urge the Roman Catholic Church to reform and welcome women into ordained ministry. Find more details at www.molleendupreedominguez.com

Today I’m talking with Katie Lacz. She works for Women’s Ordination Conference, the United States branch of a worldwide movement to urge the Roman Catholic Church to reform and welcome women into ordained ministry. Find more details at www.molleendupreedominguez.com

Silvia Laczóová pochádza z Komárna. Vyštudovala psychológiu na Univerzite Komenského v Bratislave. Pôsobila aj v USA, kde sa dostala ku koučingu, ktorému sa venuje doteraz. Je držiteľkou certifikátov: Sport Neuro Leaders a medzinárodnej federácie koučov. Svojim klientom pomáha rozdrviť bloky v mindsete, znovuzískať motiváciu a vyhnúť sa vyhoreniu. KLUB INVESTOROV https://patron.podbean.com/KLUBINVESTOROValebo na www.investiciaslovensko.sk

Audiobook. The Last Punisher: A SEAL Team THREE Sniper's True Account of the Battle of Ramadiby Kevin Lacz, Ethan E. Rocke, Lindsey LaczThis ACU Show consists of short audiobook selection from this book. Purchase this book from your favorite bookstore or Audible. Recommended by ACU.  Overview of the BookThe Last Punisher: A SEAL Team THREE Sniper's True Account of the Battle of Ramadi by Kevin Lacz, Ethan E. Rocke, Lindsey LaczDon’t miss the New York Times bestselling on-the-ground memoir from a Navy SEAL who was part of SEAL Team THREE with American Sniper Chris Kyle. Experience his deployment, from his gripping first mission to his first kill to his eventual successful return to the United States to play himself in the Oscar-nominated film directed by Clint Eastwood and starring Bradley Cooper.The Last Punisher is a bold, no-holds-barred first-person account of the Iraq War. With wry humor and moving testimony, Kevin Lacz tells the story of his tour in Iraq with SEAL Team THREE, the warrior elite of the Navy. This legendary unit, known as “The Punishers,” included Chris Kyle (American Sniper), Mike Monsoor, Ryan Job, and Marc Lee. These brave men were instrumental in securing the key locations in the pivotal 2006 Battle of Ramadi, told with stunning detail in these pages.Minute by minute, Lacz relays the edge-of-your-seat details of his team’s missions in Ramadi, offering a firsthand glimpse into the heated combat, extreme conditions, and harrowing experiences they faced every day. Through it all, Lacz and his teammates formed unbreakable bonds and never lost sight of the cause: protecting America with their fight.The Last Punisher brings the reader into the life and mind of a SEAL, demonstrating the tough realities of war. At the same time, Lacz shares how these experiences made him a better man and how proud he is of his contributions to one of this country’s most difficult military campaigns. Lacz is now an in-demand public speaker testifying to the ability of a veteran to thrive at home. The book includes an afterword on the making of the hit film American Sniper.The Last Punisher is the story of a SEAL who was never afraid to answer the call.

Denis Duboule Collège de France Évolution des génomes et développement (chaire internationale) Année 2019-2020 Régulation génétique et développement : les séquences enhancers Dans ce deuxième cours, le Pr Denis Duboule décrit les approches originales impliquant des pièges à enhancers, d’abord chez la mouche drosophile en utilisant l’élément P comme vecteur, puis chez la souris en passant par la route des cellules souches embryonnaires (ES). Il montre ensuite comment l’outil de détection principal (le gène lacZ) fut intégré dans ces stratégies expérimentales et comment ces approches ont évolué pour conduire à des stratégies actuelles plus performantes, permettant de découvrir les enhancers présent à l’intérieur d’un paysage de régulation déterminé au préalable. Se pose ensuite la question de pouvoir associer un enhancer particulier à un ou à des gène(s) cible(s) et l’importance de ce problème est illustrée par un cas de figure particulièrement parlant, à savoir la présence d’un enhancer du gène ‘gremlin’ à l’intérieur d’un gène voisin, provoquant ainsi une situation qui resta confuse pendant de nombreuses années avant d’être résolue.

Denis Duboule Collège de France Évolution des génomes et développement (chaire internationale) Année 2019-2020 Régulation génétique et développement : les séquences enhancers Dans ce deuxième cours, le Pr Denis Duboule décrit les approches originales impliquant des pièges à enhancers, d’abord chez la mouche drosophile en utilisant l’élément P comme vecteur, puis chez la souris en passant par la route des cellules souches embryonnaires (ES). Il montre ensuite comment l’outil de détection principal (le gène lacZ) fut intégré dans ces stratégies expérimentales et comment ces approches ont évolué pour conduire à des stratégies actuelles plus performantes, permettant de découvrir les enhancers présent à l’intérieur d’un paysage de régulation déterminé au préalable. Se pose ensuite la question de pouvoir associer un enhancer particulier à un ou à des gène(s) cible(s) et l’importance de ce problème est illustrée par un cas de figure particulièrement parlant, à savoir la présence d’un enhancer du gène ‘gremlin’ à l’intérieur d’un gène voisin, provoquant ainsi une situation qui resta confuse pendant de nombreuses années avant d’être résolue.

Laczó Ferenc a Maastrichti Egyetem adjunktusa beszél Pető Andreának Munkácsi Ernőről, a Zsidó Tanács prominens figurájáról, akinek most jelent meg How it Happened. Documenting the Tragedy of Hungarian Jewry, (McGill-Queen University Press, 2018) címmel angolul a naplója. Hogyan értékeli utólag írt naplójában saját és a Zsidó Tanács szerepét, mikor kezdődött az üldöztetés, és szerinte mi volt a magyar hatóságok szerepe a deportálásokban?

Former Navy SEAL sniper and published author Kevin Lacz joins Mark Zinno for episode 19 of the Kill The Quit podcast.  The guys chat about the impact that 9/11 had on them and how it lead Kevin to serve our country, his BUD/S experiences, his involvement in Clint Eastwood's movie "American Sniper", the differences between the book and move, his reason for leaving the Navy, writing "The Last Punisher", and more. 

Mark has been in hospitality from a young age, and knew early on it was what he wanted to do for his career. He's been in the business for 18 years, he's been an owner for 8.5 years. This is a great interview from a guy who worked his tail off to learn as much as he could early on, willing to do whatever it took for the short term to help set himself up for long term success. He's inspiring to listen to, he's very transparent and sincere, he offers great advice for anybody considering being an owner, he has his priorities in check, and he has big plans for his group. A great interview! Enjoy...

Episode 51 Kevin & Lindsey Lacz join me on the podcast. Kevin is a former Navy SEAL, Best Selling Author and First Class Father. His wife Lindsey was kind enough to set this up. I hope you enjoy! Thank you for making First Class Fatherhood a #1 RANKED Podcast on Apple Podcasts Charts for Kids & Family! Every Day is Father’s Day First Class Fatherhood makes an excellent Free Father’s Day Gift for any Father Figure! Support - First Class Fatherhood - Please find this Episode as well as all the Best Podcasts from First Class Fatherhood Apple Podcast App - itunes.apple.com/us/podcast/first-class-fatherhood/id1356567877 Podcasts for Android - anchor.fm/alec-lace OverCast - overcast.fm/itunes1356567877 Facebook - www.facebook.com/FirstClassFatherhood/ Twitter - www.twitter.com/aleclace Website - www.FirstClassFatherhood.com Email - emilysurprise2018@gmail.com You may not always fly first class, but you’re always a First Class Father --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app Support this podcast: https://anchor.fm/alec-lace/support

War is hell and the people who fight often have a hard time coming home. Kevin Lacz didn’t. Lacz is a former U.S. Navy SEAL who served alongside Chris Kyle in the battle of Ramadi. He survived, came home, and thrived. He’s written books, spoken to crowds, and even played himself in Clint Eastwood’s American Sniper.This week on War College, Lacz tells us what it’s like to go to war, what it’s like to want to kill people, and how to turn off the warrior when you come home.You can listen to War College on iTunes, Stitcher, Google Play or follow our RSS directly. You can reach us on our new Facebook page: https://www.facebook.com/warcollegepodcast/; and on Twitter: @War_College. See acast.com/privacy for privacy and opt-out information.

Recorded earlier - 9/19/16 In this episode of The Willie Robertson Podcast, Willie and Johnny D are joined by former SEAL Team THREE sniper, breacher and combat medic Kevin Lacz to talk about his new book The Last Punisher. The book explores Lacz's deployment with SEAL Team THREE and his experiences during the Iraq War with the legendary unit known as "The Punishers," including Chris Kyle (American Sniper), Mike Monsoor, Ryan Job, and Marc Lee. These brave men were instrumental in securing the key locations in the pivotal 2006 Battle of Ramadi, the events of which are portrayed in the Oscar-nominated film American Sniper, in which Lacz played himself. Listen to the podcast to hear what led Kevin to join the military, what advice Bradley Cooper gave him while working on American Sniper, and how experiencing failure helps make you successful.

Susan Lacz, CEO of Ridgewells Catering, speaks with Our Town host Andy Ockershausen about her life and career in Our Town. Hers is a lively interview! By the end, you will understand, if you don't already, why she has been, and continues to be, a successful business woman in Our Town. Susan has lived and worked in and around the Washington DC metro for almost 40 years. Originally from North Jersey, “totally a Jersey girl”, she moved to Arlington in 1978 to attend Marymount University which, at the time, was an all girls school. She has nothing but wonderful things to say about her college experience, and now serves on Marymount’s board of trustees. Susan attributes much of her success to her parents’ strong work ethic. Through high school, Susan worked Saturday and Sundays and during the week after school at Wyckoff Deli in Wyckoff, New Jersey. When she came to Our Town to attend college she worked at Tino’s restaurant in Arlington, just for the fun of it. She tells Andy she was hooked on the food industry from day one at Wyckoff Deli, and her passion for entertaining came from her Polish roots, more specifically her Grandmother, and upbringing. After college, Susan pursued a career in sales with Wang computers. She really enjoyed selling and was good at it. What lacked was the same passion for it that she had for food and entertaining. So she went about finding a career that captured it all - food, entertaining, sales, and passion. Susan knew she wanted to work for Ridgewells but lacked the experience they were looking for at the time, so she went to work at Clyde's at Tyson’s Corner, as a catering manager, where she was ultimately fired. Undeterred by that, she went back to Ridgewells and convinced them (after a year and a half!) that she belong there as a member of Ridgewells’ team. After ten years with the company, Susan bought the business with her partners Jose Valado and Thomas P. Keon, currently Ridgewells' COO and President, respectively. Andy fondly refers to Ridgewells as “always a family affair”. Susan has been with Ridgewells for 30 years and has owned and operated it together with her partners for 20. Ridgewells has grown into a 40 million dollar company and is considered, by the Washington Business Journal, the largest caterer in Our Town. Andy and Susan talk about competition from other caterers. Susan says that only serves to inspire them to work harder and continue to strive to be the very best. Giving back to the community is important to Susan. She makes every effort to lead her team out into Our Town community and has set Ridgewells apart from the rest. Susan serves on numerous boards including Marymount, Junior Achievement, the Committee for the Hall of Fame, Don Bosco Cristo Rey, and the Greater Washington Board of Trade. Through Ridgewells' work in the community, Susan has set an example for other caterers to do the same. Susan doesn't miss a beat as she and Andy discuss the role of women in business in Our Town. She has served on boards in the past when she was only one of two or three women sitting around the board table surrounded by "suits". She is thrilled that times have changed and more and more women are visible business leaders in Our Town. She strongly believes that women should support other women. Susan loves seeing more women take top positions in this type of business these days. She surrounds herself with positive females within her own company and gains strength in that support network. Susan is proud to be an honoree of the 2004 Washington Business Journal Women Who Mean Business program. Andy comments on the quality and service that Ridgewells is known for throughout the DC area. The servers are familiar with their customers which instills continuity and quality that people can rely on time and time again. Andy inquires about Ridgewells unique and compelling advertising. Susan explains that a few years ago,

The Total Tutor Neil Haley will interview New York Times Best Selling Author KEVIN LACZ  of American Sniper. Lacz, one of the key members to serve in the legendary unit known as "The Punishers" led by Chris Kyle whom Kevin served with side-by-side on two tours, was instrumental in securing the strategic locations instrumental in the pivotal 2006 Battle of Ramadi depicted in"American Sniper" opposite Bradley Cooper.   

The Total Tutor Neil Haley will interview  "American Sniper" Star KEVIN LACZ. He Will Discuss 9/11 and his book. Lacz, one of the key members to serve in the legendary unit known as "The Punishers" led by Chris Kyle whom Kevin served with side-by-side on two tours, was instrumental in securing the strategic locations instrumental in the pivotal 2006 Battle of Ramadi depicted in"American Sniper" opposite Bradley Cooper.  As you will recall, his NYT bestselling memoir,THE LAST PUNISHER (Simon & Schuster, ISBN #978-1-5011-2724-3) delved into America's initial plan responding to the worst terrorist attack in U.S. history and lays out challenges ahead.  

The Total Tutor Neil Haley will interview Kevin Lacz Author of THE LAST PUNISHER.   Experience his deployment, from his first mission to his first kill to his eventual successful return to the United States. THE LAST PUNISHER brings the reader into the life and mind of a SEAL, demonstrating the tough realities of war. At the same time, Lacz shares how these experiences made him a better man and how proud he is of his contributions to one of this country's most difficult military campaigns.  THE LAST PUNISHER By Kevin Lacz On-sale July 12, 2016 ISBN 9781501127243 / Hardcover / $28.00 ISBN 9781501127250 / E-book / $14.99 http://www.kevinlacz.com/   ABOUT THE AUTHOR: KEVIN LACZ left college and joined the US Navy following the terrorist attacks of September 11, 2001. He served with SEAL Team THREE as a sniper, breacher, and medic and did two combat deployments in support of Operation Iraqi Freedom. He received a Bronze Star with Valor, two Navy and Marine Corps Commendation medals in addition to others for his service. Kevin served as a technical advisor and played himself in Clint Eastwood's Oscar-nominated film American Sniper. He lives in Florida with his wife Lindsey and their two children.  

LAN Associates, a full service, engineering, planning, architecture, and surveying firm, recently celebrated its 50th Anniversary. Founded in Paterson in 1965 by John A. Lacz and now based in Midland Park, NJ, LAN Associates employs nearly 80 people.  Since its creation, LAN Associates has completed over 29,000 projects throughout the region and works on over $100 million dollars of construction projects each year for municipal, educational, hospitality, ecclesiastical, senior living, and private clients. LAN Associates service a variety of federal, state, county, and municipal governments, housing authorities, boards of education, Fortune 500 companies, foreign companies, religious facilities, and private clients. LAN’s main office is located in Midland Park, Bergen County, New Jersey; with a second location in Goshen, Orange County New York and a satellite office Trumbull, Connecticut.Its most recent high profile project was the design of the James Monroe Elementary School in Edison that was destroyed in a 2013 fire.Today on “Live From Trenton” we will learn about trends in building in New Jersey, architectural changes over the years and the challenges posed in dealing with major projects with municipalities, churches, schools and private companies as we speak with LAN Associates President Kenneth Karle. He is in charge of architectural design and architectural engineering and is in charge of master planning, conceptual design, feasibility planning, and project management for all major architectural projects for LAN Associates.

LAN Associates, a full service, engineering, planning, architecture, and surveying firm, recently celebrated its 50th Anniversary. Founded in Paterson in 1965 by John A. Lacz and now based in Midland Park, NJ, LAN Associates employs nearly 80 people.  Since its creation, LAN Associates has completed over 29,000 projects throughout the region and works on over $100 million dollars of construction projects each year for municipal, educational, hospitality, ecclesiastical, senior living, and private clients. LAN Associates service a variety of federal, state, county, and municipal governments, housing authorities, boards of education, Fortune 500 companies, foreign companies, religious facilities, and private clients. LAN’s main office is located in Midland Park, Bergen County, New Jersey; with a second location in Goshen, Orange County New York and a satellite office Trumbull, Connecticut.Its most recent high profile project was the design of the James Monroe Elementary School in Edison that was destroyed in a 2013 fire.Today on “Live From Trenton” we will learn about trends in building in New Jersey, architectural changes over the years and the challenges posed in dealing with major projects with municipalities, churches, schools and private companies as we speak with LAN Associates President Kenneth Karle. He is in charge of architectural design and architectural engineering and is in charge of master planning, conceptual design, feasibility planning, and project management for all major architectural projects for LAN Associates.

Background: Standardized and well-characterized genetic building blocks are a prerequisite for the convenient and reproducible assembly of novel genetic modules and devices. While numerous standardized parts exist for Escherichia coli, such tools are still missing for the Gram-positive model organism Bacillus subtilis. The goal of this study was to develop and thoroughly evaluate such a genetic toolbox. Results: We developed five BioBrick-compatible integrative B. subtilis vectors by deleting unnecessary parts and removing forbidden restriction sites to allow cloning in BioBrick (RFC10) standard. Three empty backbone vectors with compatible resistance markers and integration sites were generated, allowing the stable chromosomal integration and combination of up to three different devices in one strain. In addition, two integrative reporter vectors, based on the lacZ and luxABCDE cassettes, were BioBrick-adjusted, to enable beta-galactosidase and luciferase reporter assays, respectively. Four constitutive and two inducible promoters were thoroughly characterized by quantitative, time-resolved measurements. Together, these promoters cover a range of more than three orders of magnitude in promoter strength, thereby allowing a fine-tuned adjustment of cellular protein amounts. Finally, the Bacillus BioBrick Box also provides five widely used epitope tags (FLAG, His(10), cMyc, HA, StrepII), which can be translationally fused N- or C-terminally to any protein of choice. Conclusion: Our genetic toolbox contains three compatible empty integration vectors, two reporter vectors and a set of six promoters, two of them inducible. Furthermore, five different epitope tags offer convenient protein handling and detection. All parts adhere to the BioBrick standard and hence enable standardized work with B. subtilis. We believe that our well-documented and carefully evaluated Bacillus BioBrick Box represents a very useful genetic tool kit, not only for the iGEM competition but any other BioBrick-based project in B. subtilis.

Vor Beginn dieser Arbeit war ein A. fumigatus-Protein (Chp: CipC homologes Protein) mit unbekannter Funktion und hoher Homologie zum CipC-Protein aus A. nidulans als prominentes hyphenspezifisches Protein identifiziert worden (Schwienbacher, 2005). Weiterhin gab es zu diesem Zeitpunkt Hinweise, dass ein zu CipC homologes Protein in C. neoformans eine wichtige Rolle während der Virulenz spielt (Steen et al., 2003). In dieser Arbeit sollte die biologische Funktion des pilzspezifischen Proteins genauer untersucht werden. Zu diesem Zweck wurden monoklonale Antikörper gegen Chp, mehrere Reporterstämme sowie eine Deletionsmutante hergestellt. Die erhobenen Daten zeigen, dass Chp als Monomer im Cytosol der Hyphen vorliegt. Dabei zeigte sich eine gleichmäßige Verteilung eines GFP-Fusionsproteins innerhalb der Hyphen; nur die Vakuolen schienen ausgespart. Die Identifikation des Proteins auf der Sporenoberfläche von A. fumigatus (Asif et al., 2006) wurde wiederlegt und die differentielle Expression des Proteins bestätigt. Anders als in A. nidulans (Melin et al., 2002) wirkt das Antibiotikum Concanamycin A auf die Bildung von Chp in A. fumigatus nicht induzierend. Da diese Tatsache sowohl für die Na-mensgebung von CipC, als auch für die Namensgebung von Chp verantwortlich war, sollte das A. fumigatus-Protein umbenannt werden. Die Wahl des Namens fiel auf NrpA (Nitrogen regulated protein A), da die Bildung des Proteins von der N-Quelle abhängig ist. Die N-abhängige Regulation war für ein homologes F. fujikuroi Gen auf RNA-Ebene bereits bekannt (Teichert et al., 2004). In der vorliegenden Arbeit konnte sie in A. fumigatus und erstmals auf Proteinebene bestätigt werden. Desweiteren wurden auch neue N-Quellen untersucht. Dabei zeigte sich, dass NrpA in Anwesenheit der N-Quellen Glutamat, Nitrat oder Harnstoff nicht gebildet wird, wohin-gegen Komplexmedien sowie die N-Quellen Ammonium, Glutamin, Asparaginsäure, Asparagin, Valin und Tryptophan zur Bildung des Proteins führen. In Kombination einer induzierenden und einer unterdrückenden N-Quelle dominiert stets die induzierende. In Reporterstämmen (gfp; lacZ) fand diese negative Regulation der Bildung von NrpA nicht statt. Das Protein wurde sowohl in Anwesenheit einer normalerweise unterdrückenden N-Quelle, als auch in den Sporen gebildet. Weiterhin nimmt die gebildete Menge von NrpA sowohl bei längeren Inkubationszeiten, als auch bei Verwendung höherer Animpfdichten zu. Wird der Pilz zunächst mit einer die NrpA-Bildung unterdrückenden N-Quelle angezogen und dann in Medium mit einer induzierenden N-Quelle umgesetzt, dauert es 6 h bis eine Bildung von NrpA verzeichnet werden kann. Diese Zeitspanne blieb auch in einem inversen Experiment gleich. Als nächstes wurde untersucht, ob NrpA in A. fumigatus unter Stressbedingungen von Bedeutung ist. Dabei konnte gezeigt werden, dass sowohl osmotischer Stress, als auch oxidativer Stress, der durch Menadion verursacht wird, kei-ne Auswirkung auf die gebildete NrpA-Menge hat. Dagegen führt durch H2O2 verursachter Stress zu einem veränderten Laufverhalten von NrpA in SDS-Gelen. Das Protein scheint unter diesen Umständen ein höheres Molekulargewicht zu haben. Mithilfe eines A. fumigatus-Reporterstammes, der ein NrpA-GFP-Fusionsprotein bildet, konnte gezeigt werden, dass H2O2 auch zu einer veränderten Lokalisation von NrpA führt. Das sonst gleichmäßig in den Hyphen verteilte Protein formierte sich in punktförmigen Strukturen. Auch unter Mangelbedingungen spielt NrpA keine wichtige Rolle, denn weder ein C- noch ein N-Mangel verändert die gebildete Menge des Proteins. Dient die normalerweise die NrpA-Bildung induzierende N-Quelle Glutamin als C- und N-Quelle wird NrpA nicht gebildet. Ebenso wie in F. fujikuroi (Teichert et al., 2002) wird die Bildung des NrpA-Proteins durch MSX, einem Inhibitor der Glutaminsynthetase, fast vollständig inhibiert. Anders als in F. fujikuroi (Teichert et al., 2006) verursachte die Inhibierung der TOR-Kinase durch Rapamycin keinen Effekt auf die Bildung von NrpA. Auch durch eine her-gestellte Deletionsmutante konnte die biologische Funktion von NrpA nicht geklärt werden. In zahlreichen vergleichenden Untersuchungen verhielt sich die Mutante ebenso wie der Wildtyp. Der einzige dokumentierte Unterschied zwischen Mutante und Wildtyp ist eine verstärkte Bil-dung der Katalase 1 in der Deletionsmutante. Anders als angenommen spielt NrpA während der Virulenz von A. fumigatus keine Rolle. In einem Virulenzmodell in embryonierten Hühnereiern verhielten sich Deletionsmutante und Wildtyp gleich. Auch in murinen Makrophagen führten die Deletionsmutante und der Wildtyp etwa zu vergleichbaren Mengen an ausgeschüttetem IL-10 und TNFα. Ein potentieller Nutzen von NrpA bei der Diagnose der allergischen bronchoalveolaren Aspergillose (ABPA) konnte ebenso ausgeschlossen werden. Neben NrpA waren im Vergleich der Proteinmuster der verschiedenen A. fumigatus-Morphotypen auch weitere differentiell exprimierte Proteine aufgefallen. Eines davon war eine MnSOD (Aspf6), die bis dahin auch als mitochondriale MnSOD bezeichnet wurde (Rementeria et al., 2007). Da im Genom von A. fumigatus aber eine weitere MnSOD kodiert ist, die über eine putative Mitochondriensignalsequenz verfügt, sollte in einem zweiten Teil der Arbeit die tat-sächliche Lokalisation dieses Proteins gezeigt werden. Dafür wurden zunächst monoklonale Antikörper gegen das Protein hergestellt. Eine mitochondriale Lokalisation der MnSOD mit puta-tiver Signalsequenz konnte gezeigt werden. Dabei wurden sowohl Western-Blots als auch ein A. fumigatus-GFP-Reporterstamm verwendet. Weiterhin konnte das Protein genauer charakteri-siert werden. Im Gegensatz zu Aspf6, das nur in den Hyphen des Pilzes zu finden ist, wurde die mitochondriale MnSOD sowohl in den Sporen, als auch in den Hyphen nachgewiesen. Die gebil-dete Menge des Proteins verändert sich im Zeitverlauf nicht. Lediglich zu sehr späten Wachs-tumszeitpunkten in der späten stationären Phase war ein leichter Anstieg zu beobachten. Die gebildete Menge des Proteins hing auch nicht von der Animpfdichte der Kultur ab. Anders als Aspf6, das bekannterweise ein Homotetramer bildet (Flückiger et al., 2002), scheint die mitochondriale MnSOD als Dimer vorzuliegen. Auch in Antwort auf oxidativen Stress verhielten sich die beiden MnSODs unterschiedlich. Menadion, das innerhalb der Zelle die Bildung von Su-peroxidanionen bewirkt, führte zu einem leichten Anstieg der Proteinmenge der mitochondrialen MnSOD, während Aspf6 unverändert blieb. Als Folge von oxidativem Stress, der durch H2O2 verursacht wird, zeigte Aspf6 eine leichte Verringerung, während die mitochondriale MnSOD schnell abgebaut wird. In einem dritten Teil dieser Arbeit wurde die Rolle der Atmung während der Auskeimung von A. fumigatus genauer untersucht. Dabei konnte gezeigt werden, dass die Proteinbiosynthese für den Auskeimungsprozess unbedingt notwendig ist. Desweiteren wurde mit Hilfe unterschiedli-cher Methoden eine sehr frühe Aktivierung der Atmungskette während des Auskeimungspro-zesses nachgewiesen. Ebenso konnte gezeigt werden, dass die Anwesenheit von Sauerstoff für das Wachstum von A. fumigatus unbedingt erforderlich ist. Im anaeroben Milieu konnten die Konidien weder anschwellen noch auskeimen. Auch bereits vorhandene Hyphen konnten unter Abwesenheit von Sauerstoff nicht weiterwachsen. Weitere Untersuchungen zeigten, dass A. fu-migatus anders als A. nidulans (Takasaki et al., 2004) nicht über die Fähigkeit verfügt, im Anae-roben durch eine Fermentation von Ammonium zu überleben. In dieser Arbeit wurde ebenso wie in anderen aktuellen Arbeiten (Williger et al., 2008) die Fähigkeit von A. fumigatus, in hypoxischen Umgebungen zu wachsen, nachgewiesen.

Fibroblast growth factor-23 (FGF-23), a molecule that is mutated in patients with autosomal dominant hypophosphatemic rickets (ADHR), appears to be involved in the regulation of phosphate homeostasis. Although increased levels of circulating FGF-23 were detected in patients with different phosphate-wasting disorders, it is not yet clear whether FGF-23 is directly responsible for the abnormal regulation of mineral ion homeostasis and consequently bone development. To explore these questions further, we generated a mouse model in which the entire Fgf-23 gene was replaced with the lacZ gene. Fgf-23 null (Fgf-23-/-) mice showed signs of growth retardation, developed severe hyperphosphatemia with elevated serum 1,25(OH)2D3 levels, and died by 13 weeks of age. Hyperphosphatemia in Fgf-23-/- mice was accompanied by skeletal abnormalities, as demonstrated by histological, molecular, and morphometric analyses. Fgf-23-/- mice had decreased bone mineral density of the limbs due to an accumulation of unmineralized osteoid leading to marked limb deformities. Moreover, Fgf-23-/- mice showed excessive mineralization in soft tissues. To further expand our understanding regarding the role of Fgf-23 in phosphate homeostasis and skeletal mineralization, we crossed Fgf-23-/- animals with Hyp mice, the murine equivalent of XLH. Interestingly, Hyp males lacking both Fgf-23 alleles were indistinguishable from Fgf-23-/- mice, both in terms of serum phosphate levels and skeletal changes, suggesting that Fgf-23 is downstream of the phosphate regulating gene with homologies to endopeptidases on the X chromosome (Phex). To explore further the role of the vitamin D axis for FGF-23 signaling, we mated Fgf-23 deficient mice and vitamin D receptor (VDR) mutant mice with a non-functioning VDR. To prevent secondary hyperparathyroidism in VDR and compound mutant mice, all mice were kept on a rescue diet enriched with calcium, phosphorus, and lactose. In analogy to previous findings, Fgf-23-/- animals showed hypercalcemia, hyperphosphatemia, growth retardation, ectopic calcifications, severe osteoidosis, skin atrophy, and renal dysfunction. In addition, here we describe that Fgf-23-/- mice are hypoglycemic, and have profoundly increased peripheral insulin sensitivity and improved subcutaneous glucose tolerance, but normal renal expression of the aging suppressor gene Klotho. Although VDR and double mutants on the rescue diet still had moderately elevated parathyroid hormone serum levels and lower bone mineral 76 density compared to wild-type mice, double mutant mice had normal body weight, were normocalcemic and normophosphatemic, and ectopic calcifications as well as renal dysfunction were absent. Ablation of vitamin D signaling in compound mutants also normalized subcutaneous glucose tolerance tests and insulin secretory response. In conclusion, our results indicate that the alterations in mineral and carbohydrate metabolism present in Fgf-23-/- mice require an intact vitamin D signaling pathway.

The BCR/ABL1 fusion protein is found in virtually all cases chronic myeloid leukemia (CML) and a large proportion of acute lymphoblastic leukemia (ALL). The fact that the BCR/ABL1 fusion protein is crucial for the development of leukemia makes this fusion protein an attractive target for therapy development. We have developed a strategy for the in vivo detection of the BCR/ABL1 fusion protein, in which the presence of the BCR/ABL1 fusion protein is detected intracellularly and if the fusion protein is present an arbitrary action is initiated in the cell (e.g. mark the cells or selectively kill the cells). Our BCR/ABL1 detection strategy is based on protein-protein interactions. Two detection proteins are expressed in the cells: 1) protein A, a GAL4-DNA binding domain/BCR interacting protein fusion protein (GAL4DBD-BAP-1) and 2) protein B, a GAL4-activation domain/ABL interacting protein fusion protein (GAL4AD-CRKL). Only when BCR/ABL1 is present in the cell, do protein A, protein B, and BCR/ABL1 form a trimeric complex which activates the transcription of reporter genes under the control of GAL4-upstream activating sequence (UAS). A proof of principle for the strategy was implemented in the yeast system. We did not use full length BAP-1 or CRKL but only those portions of the proteins that directly interacted with BCR or ABL, respectively. We showed in the yeast two hybrid system, that the C-terminus of BAP-1(amino acids 617-879) binds to full length BCR. The site of interaction of CRKL and ABL was confirmed to be the N-terminal SH3 domain (SH3n) of CRKL as described in the literature. Yeast cells (strain CG1945) transformed with a protein A expressing plasmid (pGBT9-BAP), a protein B expressing plasmid (pGAD424-CRKLSH3n), and a BCR/ABL expressing plasmid (pES1/BCR-ABL) showed expression of the reporter genes HIS3 and LACZ. The expression of the HIS3 reporter gene was assayed by growth of the yeast cells on medium lacking histidine. The expression of the LACZ gene was verified by a beta-galactosidase filter assay. Yeast cells that were transformed with the pES1 plasmid without the BCR/ABL1 coding region did not show activation of the reporter genes. Several other negative controls demonstrated the specificity of the assay. Thus the method was able to clearly distinguish between BCR/ABL expressing cells and cells did not express BCR/ABL1. We then adapted this system for use in mammalian cells. The open-reading frames encoding the proteins A and B were recloned into mammalian expression vectors. The human embryonal kidney cell line HEK293 and the murine myeloid progenitor cell line 32D which had been stably transfected with a BCR/ABL expressing plasmid were tested. The firefly luciferase gene and the yellow fluorescent protein (eYFP) were used to evaluate the whole cell population and single cell, respectively. Unfortunately, the system failed to work in the mammalian cell lines tested. Even though the detection system did not work in mammalian cells, most likely due to the cytoplasmic localization of the BCR/ABL1 fusion protein, it should still be a viable strategy for the detection of leukemia-associated fusion protein, which localize to the nucleus (i.e AML-ETO). This strategy could be adapted for purging the bone marrow of leukemia patients using therapeutically more useful effector genes like suicide genes, which encode pro-drug converting enzymes (e.g. HSV thymidine kinase), or markers that can easily be assayed (e.g. YFP).

In der vorliegenden Arbeit wurden die immunogenen Eigenschaften des Nichtstruktur-proteins 3 (NS3) des Virus der Bovinen Virusdiarrhö (BVDV) erstmals im natürlichen Wirt charakterisiert. Hierzu wurden sechs Kälber dreimal im Abstand von vier Wochen mit BVDV-NS3-rekombinantem modifiziertem Vacciniavirus Ankara (MVA) intramuskulär immunisiert (>108 infektiöse Einheiten). Vier Kälber, die mit LacZ-rekombinantem MVA (>108 infektiöse Einheiten) immunisiert wurden, bildeten die Kontrollgruppe. Fünf Wochen nach der letzten Immunisierung erfolgte eine Belastungsinfektion mit dem BVD-Virusisolat PT810 (106 kulturinfektiöse Dosen50). Für die Untersuchung der BVDV-spezifischen zellulären Immunitätsmechanismen wurden die Lymphozyten aller Tiere mit infektiösem BVDV-PT810 in vitro restimuliert und mit Hilfe verschiedener Testsysteme untersucht. Eine zytofluorometrische Nachweismethode (Fluoreszenzfarbstoff: Carboxyfluorescein Diacetate Succinimidyl Ester [CFSE]) diente der Detektion einer antigenspezifischen Lymphozytenproliferation. Die Aktivität BVDV-spezifischer zytotoxischer T-Lymphozyten (CTL) gegen autologe, BVDV-infizierte Zielzellen wurde mit Durchflusszytometrie gemessen. Mit Hilfe einer Real Time PCR wurde die Interleukin-2 (IL-2) und 4 (IL-4) mRNA-Synthese untersucht. Ein kommerziell erhältlicher ELISA (Bovigam) diente dem Nachweis der γ-Interferon-Synthese. Über einen Zeitraum von 19 Tagen nach der intranasalen BVDV-Testinfektion wurden die Kälber täglich klinisch untersucht, die Gesamtleukozytenzahl im peripheren Blut bestimmt, quantitative BVDV-Isolierungen aus den Leukozyten durchgeführt und BVDV-spezifische Antikörper mit Serumneutralisationstesten gemessen. Bereits vor der Belastungsinfektion ließ sich bei allen Kälbern, die mit BVDV-NS3-rekombinatem MVA immunisiert wurden, eine antigenspezifische Lymphozytentransformation nachweisen. Ebenso konnte bei vier Tieren der Versuchsgruppe (N = 6) nach der dritten Immunisierung eine deutliche BVDV-spezifische Zytotoxizität gegenüber autologen Hodenzellen detektiert werden. Nach der BVDV-Belastungsinfektion war die spezifische Zytotoxizität bei fünf der sechs BVDV-NS3-MVA-Impftiere höher als bei den Tieren der LacZ-Kontrolle. Im Gegensatz zur Kontrollgruppe ließ sich bei der Versuchsgruppe nach der dritten Immunisierung ein deutlicher Anstieg der IL-2 und IL-4 mRNA-Syntheserate von in vitro mit BVDV-restimulierten Lymphozyten nachweisen. Bei fünf der sechs Kälber war zu diesem Zeitpunkt auch γ-Interferon detektierbar. Der Nachweis von BVDV-neutralisierenden Antikörpern war vor der Testinfektion bei keinem Kalb möglich. Nach der Testinfektion konnte ein spezifischer Priming-Effekt für alle immunologischen Parameter, einschließlich der Bildung BVDV-neutralisierender Antikörper, festgestellt werden. Erwartungsgemäß wurden bei beiden Tiergruppen keine Krankheitssymptome beobachtet. Das Ausmaß der Leukopenie infolge der Testinfektion war gleich. Dagegen war der semiquantitativ bestimmte BVDV-Titer in Blutproben der Kontrollgruppe an den Tagen 3-13 nach der Infektion etwa doppelt so hoch (1,9-fach) wie bei der Versuchsgruppe (p = 0,016; signifikant). Im Mittel war bei der Versuchgruppe an 6,7 Tagen und bei der Kontrollgruppe an 8,0 Tagen BVDV reisolierbar (p = 0,121; nicht signifikant). Neben den beachtlichen immunologischen Stimulationseffekten wurde die Reduktion der Viruslast als Hinweis für eine protektive Wirkung des BVDV-NS3 gewertet.

The transcriptional activator CadC in Escherichia coli, a member of the ToxR-like proteins, activates transcription of the cadBA operon encoding the lysine decarboxylase CadA and the lysine-cadaverine antiporter CadB. cadBA is induced under conditions of acidic external pH and exogenous lysine; anoxic conditions raise the expression level up to 10 times. To characterize the binding mechanism of CadC, procedures for the purification of this membrane-integrated protein and its reconstitution into proteoliposomes were established. The binding sites of CadC upstream of the cadBA promoter region were determined by in vitro DNaseI protection analysis. Two regions were protected during DNaseI digestion, one from - 144 to - 112 bp, designated Cad1, and another one from - 89 to - 59 bp, designated Cad2. Binding of purified CadC to Cad1 and Cad2 was further characterized by DNA-binding assays, indicating that CadC was able to bind to both DNA fragments. Genetic analysis with promoter-lacZ fusions confirmed that both sites, Cad1 and Cad2, are essential for activation of cadBA transcription. Moreover, these experiments revealed that binding of H-NS upstream of the CadC-binding sites is necessary for repression of cadBA expression at neutral pH and under aerobic conditions. Based on these results, a model for transcriptional regulation of the cadBA operon is proposed, according to which H-NS is involved in the formation of a repression complex under non-inducing conditions. This complex is dissolved by binding of CadC to Cad1 under inducing conditions. Upon binding of CadC to Cad2 cadBA expression is activated. Copyright (C) 2005 S. Karger AG, Basel.

The hyperpolarization-activated, cyclic-nucleotide-gated cation channel (HCN) familiy comprises four members. The channels play a role in the formation of rhythmic activity of heart and brain. In contrast to the other three members of the HCN channel family, there are only a few studies of tissue distribution and electrophysiological properties of HCN3 so far. Expression has been reported at low levels, but throughout the brain, in some other tissues such as retina, olfactory epithelium and recently in the heart. In order to study the physiological relevance of HCN3 expression, we generated HCN3-deficient mouse lines by gene targeting and homologous recombination: a complete Knockout, a complete Knockout expressing the reporter gene lacZ instead of HCN3 and a conditional Knockout using the Cre-loxP system to study spatial and temporal functions of this pacemaker channel. The functional characterization included behavioural studies and heart physiology.

Im Rahmen der vorliegenden Doktorarbeit wurde zum ersten Mal die Keimbahn eines Käfers erfolgreich genetisch transformiert. Der von uns zu diesem Zweck in Zusammenarbeit mit Ernst Wimmer entwickelte Transformationsmarker 3xP3-EGFP hat inzwischen sein Potential als spezies-unabhängiges Markergen auch in weiteren Invertebraten-Spezies unter Beweis gestellt und damit das Spektrum transformierbarer Taxa beträchtlich erweitert. In Tribolium konnten Transformationsereignisse mit 3xP3-EGFP für drei verschiedene Transposons - Hermes, Minos und piggyBac - erzielt werden. Die Effizienz betrug dabei 1,4% (Hermes), 11,4% (Minos) bzw. 56% (piggyBac) der fertilen G0 und gehört damit zu den höchsten Werten, die in der Literatur für Insekten berichtet wurden. Bei Minos konnte die Effizienz durch die Verwendung von Transposase mRNA statt einem DNA Helper-Plasmid weiter auf 32,4% gesteigert werden. Für piggyBac und Minos wurde ferner in Zusammenarbeit mit anderen Labors gezeigt, daß es sich bei den meisten Transposoninsertionen um unabhängige Einzelintegrationen handelt, die auf verschiedene Chromosomen verteilt sind und stabil weitervererbt werden. Die Größe zusätzlich transferierter Fremd-DNA kann dabei bei piggyBac mindestens bis zu 9,5 kb betragen. Schließlich konnte noch ein piggyBac Element durch Helperinjektion mit einer Rate von 28,1% remobilisiert werden. Zusammen mit der Anfälligkeit für enhancer trap Effekte können daher mit diesem System alle relevanten Transposon-basierenden Techniken zur funktionellen Genomanalyse angewandt werden. Als erste praktische Anwendung wurden D. melanogaster Sequenzen für anteriore und posteriore mRNA-Lokalisierung (bicoid-3’UTR und oskar-3’UTR), sowie ein bicoid-abhängiger Minimalpromotor in Tribolium eingeführt. Allerdings konnten durch diese Ansätze keine Komponenten oder Mechanismen eines ggf. konservierten maternalen Systems nachgewiesen werden. Ein Konstrukt mit 5,2 kb der upstream Sequenzen von Tc’hunchback mit lacZ als Reportergen war hingegen in der Lage, das endogene hunchback-Muster größtenteils nachzubilden. Das frühere Ergebnis von Christian Wolff mit Tc’hunchback in Drosophila, wonach dieses Fragment alle wesentlichen regulatorischen Elemente enthält, konnte daher in transgenen Käfern bestätigt werden. Zusätzlich zu dem als sehr riskant eingestuften Transformations-Projekt wurde parallel ein weiteres Projekt durchgeführt, die Analyse der homöotischen Mutanten wurm und überlänge. In beiden Mutanten ist vor allem die Identität der posterioren Segmente ab A9 verändert. In wurm sind die Segmente A9-A11 nach A8 transformiert und die telsonalen Anhänge Urogomphi und Pygopodien fehlen. In überlänge ist nur A9 wie A8 ausgebildet und demzufolge nicht mit dem Telson fusioniert. Es fehlen nur die Urogomphi. überlänge bildet zusätzlich ein ektopisches Stigma im ersten thorakalen Segment. Es wurde gezeigt, daß es sich bei den betroffenen Genen um zwei verschiedene Loci handelt, die beide nicht im homöotischen Komplex liegen. Obwohl der Phänotyp von wurm weitgehend der RNAi-Phänokopie von Abdominal-B entspricht, konnte also keiner dieser beiden Loci einem bekannten Hox-Gen zugeordnet werden. Als mögliches Kandidatengen für diese Loci wurde daher das Tribolium-Homolog des regionsspezifischen homöotischen Gens spalt kloniert. Die Expression von spalt entspricht weitgehend der von Dm’spalt, mit einer anterioren und einer posterioren Domäne, einer dorsalen Expression an den seitlichen Rändern des Keimstreifs, sowie einem komplexen Muster im Nervensystem. Mit Hilfe der kürzlich entwickelten Technik der parentalen RNAi wurde die Funktion dieses Gens untersucht. In sal–– Phänokopien finden sich, wie in Drosophila, anteriore und posteriore Veränderungen von Segmentidentitäten. So wird das abdominale Segment A9 in Richtung anteriore abdominale Segmente transformiert. Dadurch tritt ein zusätzliches Stigma auf und die Pygopodien gehen verloren, das Segment fusioniert aber weiterhin mit dem Telson. Im Gegensatz zu Drosophila wird aber anterior nicht das Labium verändert, sondern die Identität der Maxille wird partiell in Richtung Mandibel transformiert: statt dem Enditen der Maxille wird ein mandibel-ähnlicher Zahn gebildet. Damit kommt offenbar auch spalt nicht als Locus in Frage, der in wurm oder überlänge seine Funktion verloren hat. Möglicherweise spielen diese beiden Loci eine Rolle als den HOX-Genen übergeordnete regulatorische Gene, oder als Co-Faktor von Abd-B. Damit sind wurm und überlänge als interessante (und aus Drosophila nicht bekannte) Spieler im homöotischen System der Insekten identifiziert, was weitere Untersuchungen als sehr lohnend erscheinen läßt. Vor allem aber hat dieses Teilprojekt die Evolution des spalt-Gens erhellt, das in weniger abgeleiteten Insekten offenbar eine essentielle Rolle bei der Spezifizierung von Mandibel versus Maxille spielt. Diese Funktion ist in Drosophila vermutlich im Zuge der Reduktion der Mandibel verloren gegangen.

Nach der Aufklärung der Basenabfolge des Genoms von Saccharomyces cerevisiae ist die Funktion der 30.000-40.000 Gene und insbesondere das Zusammenspiel der Regulation der einzelnen Gene ein zentrales Thema der Molekularbiologie. Die DNA eukaryonter Zellen liegt durch Bindungen an Histon-Proteine im Zellkern als Chromatin vor. Die Chromatinstruktur dient nicht nur der Komprimierung der DNA auf engstem Raume, sondern hat auch starke Auswirkungen auf die Funktion der DNA. So müssen Gene bei ihrer Aktivierung durch Veränderung ihrer Chromatinstruktur, die bis zur Ablösung der Histone führen kann, den für die Transkription benötigten Enzymen und Faktoren erst zugänglich gemacht werden. Das PHO5-Gen der Hefe Saccharomyces cerevisiae stellt ein sehr gut untersuchtes Modell dar, bei dem Veränderungen der Chromatinstruktur genau untersucht und mit dem funktionellen Zustand des Gens korreliert worden sind. PHO5 kodiert für eine saure Phosphatase, die bei Verbrauch der Phosphatreserven der Zelle in den periplasmatischen Raum sezerniert wird, um aus dort eventuell vorhandenen organischen Phosphatverbindungen Phosphat zu gewinnen. Ist im Medium genügend Phosphat vorhanden, ist PHO5 reprimiert. In diesem Zustand ist die Chromatinstruktur des PHO5-Promotors durch vier dicht aufeinander folgende Nukleosomen gekennzeichnet, wodurch der Promotor Enzymen und regulatorischen Proteinen allgemein schlecht zugänglich ist. Nur zwischen dem zweiten und dem dritten Nukleosom ist die dichte Anordnung der Nukleosomen durch einen etwa 70 bp langen gut zugänglichen Bereich unterbrochen. In dieser sogenannten hypersensitiven Region bindet bei Phosphatmangel der aktivierende Transkriptionsfaktor Pho4 gemeinsam mit dem Faktor Pho2 an ein UAS-Element und induziert die PHO5-Expression. Dabei lösen sich die vier Nukleosomen vom DNA-Strang ab. Sin4 ist ein Transkriptionsfaktor der Hefe Saccharomyces cerevisiae, der auf mehrere Promotoren zumeist reprimierenden Einfluss ausübt. Ausgangspunkt der hier vorliegenden Arbeit war der Befund, dass in Abwesenheit von Sin4 die Gegenwart der prokaryontischen lacZ Sequenz stromaufwärts des PHO5-Promotors zu einer Derepression des PHO5-Gens führt, und zwar in Gegenwart von Phosphat, also unter eigentlich reprimierenden Bedingungen. Dieser Effekt wurde ursprünglich bei der Verwendung der kodierenden Sequenz von lacZ als dem PHO5-Promotor nachgeschalteten Reporter-Gen in sin4-Hefezellen entdeckt. Eine Frage der hier vorliegenden Arbeit galt der Ursache der Derepression von PHO5 durch die lacZ kodierenden DNA-Sequenz. Dazu interessierte uns, ob die Derepression ein spezielles Phänomen der lacZ-Sequenz ist oder ob es sich hierbei eher um eine allgemeine Eigenschaft von DNA-Fragmenten handelt. Außerdem interessierte uns, ob die Herkunft der DNA aus prokaryonten oder eukaryonten Zellen eine Rolle spielen könnte. Dazu wurde jeweils eine große Anzahl zufällig ausgewählter DNA-Fragmente einer Länge zwischen 900bp und 1200bp aus den Genomen der Hefe Saccharomyces cerevisiae und der Bakterien Escherichia coli und Micrococcus lysodeikticus an entsprechender Stelle vor den PHO5-Promotor integriert. Die so konstruierten Plasmide wurden in einen Hefestamm transformiert, in dem das SIN4-Gen zerstört worden war. Insgesamt wurden 400 Klone mit integrierten Hefe-DNA-Fragmenten, 300 Klone mit integrierten M. lysodeikticus-DNA-Fragmenten und 14 Klone mit integrierten E. coli-DNA-Fragmenten untersucht. Die Bestimmung der Phosphatase-Aktivitäten der einzelnen Klone ergab für fast alle Plasmide mit integrierten E. coli- und M. lysodeikticus-DNA-Fragmenten eine erhöhte Aktivität trotz phosphatreichen Mediums. Im Gegensatz dazu zeigten die wenigsten Plasmide mit integrierten Hefe-DNA-Fragmenten eine Erhöhung der PHO5-Expression unter denselben Bedingungen. Von den insgesamt 400 getesteten Plasmiden wiesen nur neun eine gesteigerte PHO5-Expression auf. In allen Fällen, also für alle E. coli-, M. lysodeikticus- und Hefe-DNA-Fragmente, wurde nur in Abwesenheit von Sin4 eine erhöhte Phosphatase-Aktivität gemessen. Bei seiner Anwesenheit wurden in phosphatreichem Medium nie gesteigerte Aktivitäten beobachtet. Diese Ergebnisse zeigen deutlich, dass die hier beobachtete Derepression typischerweise eine Eigenschaft prokaryonter DNA ist. Nur ein Bruchteil der eukaryonten DNA-Fragmente aus dem Hefe-Genom führt zu einer Derepression der Promotoraktivität, während dies nahezu alle prokaryonten DNA-Fragmente aus Escherichia coli- bzw. Micrococcus lysodeikticus tun. Um die neun Hefe-DNA-Fragmente, die zu einer Aktivierung des PHO5-Promotors führten, auf eventuelle Besonderheiten zu untersuchen, wurden ihre DNA-Sequenzen bestimmt und analysiert. Außerdem wurden noch zwei E. coli-DNA-Fragmente sequenziert, die zu keiner gesteigerten PHO5-Expression geführt haben. Diese sehr eindeutigen Ergebnisse werfen Fragen nach dem zugrunde liegenden Mechanismus auf. Eventuelle DNA-Methylierungen oder kryptische Promotoren schieden als Erklärung des Phänomens aus. Unterschiede des G-C-Gehalts der einzelnen DNA-Fragmente könnten besonders für die prokaryonte DNA teilweise eine Erklärung liefern. Die beiden prokaryonten Genome haben mit 51% bzw.72% einen wesentlich höheren G-C-Gehalt als das Hefegenom mit 38%. Besonders die beiden E. coli-DNA-Fragmente, die zu keiner gesteigerten PHO5-Expression führten, besitzen einen wesentlich geringeren G-C-Gehalt als der Durchschnitt des gesamten E. coli-Genoms (44,7% bzw. 38,0% im Vergleich zu 51%). Eukaryonte DNA besitzt in ihrer Sequenz im Gegensatz zu der aus Prokaryonten eine gewisse Periodizität, die sich etwa alle 10,5bp wiederholt und die Ausbildung von Nukleosomen erleichtert. Das Fehlen dieser Periodizität in prokaryonter DNA könnte sich ebenfalls auswirken, z.B. über eine labile Chromatinstruktur, die sich auch auf den benachbarten PHO5-Promotor auswirkt und dadurch eine Dereprimierung von PHO5 in sin4-Zellen auslöst. Die Dereprimierung des PHO5-Promotors durch die wenigen Hefe-DNA-Fragmente trotz reprimierender Bedingungen könnten aufgrund anderer Mechanismen zustande zu kommen. Die neun Hefe-DNA-Fragmente, die zu einer Aktivierung des PHO5-Promotors führten, zeigten auch keinen vom Hefegenom abweichenden G-C-Gehalt. Es ist auffällig, dass alle 9 DNA-Fragmente intergenische Bereiche enthalten. In diesen Bereichen gibt es oft regulatorische Elemente, die häufig in hypersensitiven Regionen gefunden werden. Hypersensitive Regionen sind nicht in Nukleosomen gepackt und könnten dadurch auch die umgebene Chromatinstruktur beeinflussen. Unabhängig von den mechanistischen Überlegungen zeigen diese Untersuchungen, dass die Aktivität eines Promotors von der Umgebung beeinflusst werden kann und dass daher der Einsatz von heterologen Reportergenen mit Vorsicht betrachtet werden muss.

Selective expression of murine cytomegalovirus (MCMV) immediate-early (IE) genes leads to the presentation by the major histocompatibility complex (MHC) class I molecule L a of a peptide derived from MCMV IE protein pp89 (Reddehase, M. J., J. B. Rothbard, and U. H. Koszinowski. 1989. Nature (Lond.). 337:651). Characterization of endogenous antigenic peptides identified the pp89 peptide as the nonapeptide msYPHFMFFNLt76 (del Val, M., H.-J. Schlicht, T. Ruppert, M. J. Reddehase, and U. H. Koszinowski. 1991. Cell. 66:1145). Subsequent expression of MCMV early genes prevents presentation of pp89 (del Val, M., K. Mfinch, M. J. Reddehase, and U. H. Koszinowski. 1989. Cell. 58:305). We report on the mechanism by which MCMV early genes interfere with antigen presentation. Expression of the IE promoter-driven bacterial gene lacZ by recombinant MCMV subjected antigen presentation of B-galactosidase to the same control and excluded antigen specificity. The La-dependent presence of naturally processed antigenic peptides also in nonpresenting cells located the inhibitory function subsequent to the step of antigen processing. The finding that during the E phase of MCMV gene expression the MHC class I heavy chain glycosylation remained in an Endo H-sensitive form suggested a block within the endoplasmic reticulum/c/s-Golgi compartment. The failure to present antigenic peptides was explained by a general retention of nascent assembled trimolecular MHC class I complexes. Accordingly, at later stages of infection a significant decrease of surface MHC class I expression was seen, whereas other membrane glycoproteins remained unaffected. Thus, MCMV E genes endow this virus with an effective immune evasion potential. These results also indicate that the formation of the trimolecular complex of MHC dass I heavy chain, ~2-microglobulin, and the finally trimmed peptide is completed before entering the medial-Golgi compartment.