Podcasts about transfection

In this episode of Peak Human Labs, Dr. Sanjeev Goel sits down with Dr. Ryan Rossner, a former NFL player turned PhD researcher, now working at MiniCircle—a pioneering company developing the world's first reversible human genetic enhancement platform. Using highly expressive DNA plasmids, MiniCircle's innovative approach allows genes to be added to the body without altering the existing genome.   Dr. Rossner delves into groundbreaking health optimization strategies, particularly gene therapy involving the follistatin gene, which enhances muscle growth, reduces inflammation, and supports rejuvenation. The conversation expands to other gene therapies, the role of sleep and exercise in longevity, and the importance of a holistic approach to health.   They also explore cutting-edge longevity science, including hibernation's impact on metabolism, metabolic flexibility, and seasonal metabolic shifts. Dr. Rossner introduces the concept of metabolic elasticity—how an adaptable metabolism can extend lifespan. He breaks down key longevity mechanisms such as fasting, insulin regulation, and AMPK activation while highlighting promising research areas like ICAR and plasmapheresis. Throughout the discussion, he shares insights from his work at MiniCircle, offering a glimpse into the future of health and longevity interventions.   Ready to dive deeper into the latest breakthroughs in longevity and health optimization? Subscribe now, share this episode with a friend, and stay tuned for more expert insights on Peak Human Labs! Key Takeaways Longevity and health optimization Gene therapy and its mechanisms Follistatin gene therapy for body composition and rejuvenation Accessibility and cost of gene therapy treatments Benefits of increased follistatin levels, including muscle growth and reduced inflammation Lifestyle factors contributing to longevity, such as sleep, exercise, and relationships Importance of self-monitoring and health-tracking tools Role of sleep in overall health and longevity In This Episode: [00:00:00] Introduction to the podcast [00:01:17] Guest introduction – Dr. Ryan Rossner [00:03:00] Mini Circle overview [00:03:37] Follistatin gene benefits [00:04:05] Gene therapy mechanism [00:04:57] Transfection process [00:05:22] DNA integration clarification [00:05:58] Follistatin level variation [00:06:44] Therapy duration [00:07:06] Accessibility of treatment [00:07:42] Future gene therapies [00:08:09] Clotho gene benefits [00:08:47] Patient treatment outcomes [00:09:30] Biological age measurement [00:09:43] Age reduction results [00:10:19] Personal experience in longevity [00:11:46] Pyramid of longevity [00:12:02] Individual variation in treatment response [00:12:18] Importance of sleep [00:13:10] Sleep and detoxification [00:13:44] Orexin receptor antagonists [00:14:20] Impact of aging on sleep [00:16:28] Weekly exercise routine [00:17:07] Recovery practices [00:20:59] Brain health insights [00:21:33] Supplements for brain health [00:21:52] Sauna and brain health [00:22:31] Hypoxia chamber experience [00:23:31] Research on hypoxia and longevity [00:24:47] Comparing oxygen levels [00:25:13] Creatine for brain benefits [00:26:01] Vitamin D and health [00:26:57] Rapamycin in longevity [00:27:48] Episodic use of rapamycin [00:30:17] Metabolic depression and triggers [00:31:64] Metabolic elasticity [00:34:03] Seasonal metabolism [00:35:06] Underappreciated longevity mechanisms [00:35:21] Genetic pathways and longevity [00:37:03] Interest in plasmapheresis [00:38:04] Future work at Mini Circle [00:38:38] Resources for learning   Our Guest Ryan Rossner is a Ph.D. student in Molecular Medicine at the University of Washington, researching flavin-containing monooxygenase proteins in aging. Dr. Matt Kaeberlein, studies mechanisms of longevity and is a trainee in the Biological Mechanisms of Healthy Aging Program. As Director of Longevity Research at MiniCircle, he develops gene therapy platforms for health optimization. An active biohacker, he experiments with tools like altitude generators and ice baths. Rossner shares insights on aging and longevity through social media, contributing to the broader conversation on extending human potential. Resources and Links Peak Human Labs https://www.youtube.com/@peakhumanlabs/videos https://www.peakhuman.ca/ https://www.instagram.com/peakhumanlabs/?hl=en https://open.spotify.com/show/5hx9R37ElxgzCrBccRWoHd?si=8atK0n82QbeL3DWg5-vjvg&nd=1&dlsi=ce0f77aa4f304724   Dr. Sanjeev Goel https://www.linkedin.com/in/sanjeevgoelmd/?originalSubdomain=ca   Dr. Ryan Rossner   https://minicircle.io https://www.instagram.com/rjrphd/  

Protect Your Retirement W/ a Gold and/or Silver IRA: https://www.sgtreportgold.com/ or CALL( 877) 646-5347 - Noble Gold is Who I Trust   Dr. Robert Young returns to SGT Report with tangible scientific data that will blow you away. Do you know what's in the new popular diet drug that's sweeping the nation? Do you know what's in the blood of all of us now? Do you know how to get it out? Watch NOW to discover the TRUTH.   Get the NANO-TECH & Heavy Metals out of your blood w/ Master Peace https://masterpeacebyhcs.com/?ref=4094   PH Miracle Products: https://phmiracleproducts.com?aff=63   Please Support Dr. Young's Legal Fund on Give Send Go: https://www.givesendgo.com/G2Z76 https://old.bitchute.com/video/CcQPc3qEUlBU/

Joining me today is Jay Couey PhD, here to discuss the ongoing COVID-19 illusion, the many and varied dangers of the COVID injections, and the very serious risk of blindly following the consensus (scientific or otherwise) instead of considering all possibilities and coming to your own conclusions. We also review the overlap of nanotechnology with this conversation, focusing on the lipid nanoparticles within the injections, and discuss the possibility of there being more to these lipids than we have been told.  !function(r,u,m,b,l,e){r._Rumble=b,r[b]||(r[b]=function(){(r[b]._=r[b]._||[]).push(arguments);if(r[b]._.length==1){l=u.createElement(m),e=u.getElementsByTagName(m)[0],l.async=1,l.src="https://rumble.com/embedJS/u2q643"+(arguments[1].video?'.'+arguments[1].video:'')+"/?url="+encodeURIComponent(location.href)+"&args="+encodeURIComponent(JSON.stringify([].slice.apply(arguments))),e.parentNode.insertBefore(l,e)}})}(window, document, "script", "Rumble");   Rumble("play", {"video":"v4c2pcc","div":"rumble_v4c2pcc"}); Source Links: GigaohmBiological's Videos - Twitch (50) Gigaohm Biological Archive Gigaohm Biological (27)

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.26.550695v1?rss=1 Authors: Hood, E. M., Lipinski, R. A. J., Lipinski, D. M. Abstract: PURPOSE ARPE19 cells are a commonly used cell culture model for the study of retinal pigment epithelial cell biology and pathologies. However, numerous studies have demonstrated that ARPE19 undergo morphologic, transcriptomic and genomic alterations over time and with increasing passage number. Herein, we explore the mechanisms underlying increased resistance to the delivery of exogenous genetic material via transfection in ARPE19 cells using mass spectrometry. METHODS ARPE19 cells (N=5 wells/reagent) were seeded in 6-well plates at passages 24 through 30. At 70% confluency an mCherry reporter construct was delivered via transfection using Lipofectamine 3000, Lipofectamine LTX, Lipofectamine Stem, or PEI (polyethylenimine) reagents. After 72 hours, transfection efficiency was quantified by fluorescence microscopy and flow cytometry. Mass spectrometry and immunofluorescence of ARPE19 cells were performed at passages 24 and 30 to evaluate altered protein synthesis and localization between passage numbers. RESULTS ARPE19 transfection showed a maximum transfection efficiency of 32.4% at P26 using Lipofectamine 3000 reagent. All lipofectamine based reagents demonstrated statistically significant decreases in transfection efficiency between passages 24 and 30. Mass spectrometry analysis revealed 18 differentially expressed proteins, including down-regulation of clathrin light chain B (CLTB) and legumain (LGMN) that was confirmed via immunofluorescence imaging, which indicated altered intracellular localization. CONCLUSIONS ARPE19 cells demonstrate passage number dependent changes in lipofectamine-based transfection efficiency. Mass spectrometry and immunofluorescence indicates the observed decrease in transfection efficiency involves the dysregulation of endocytosis and intracellular endolysosomal trafficking at later passages. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

0:00 Into 10:30 Insane News 46:59 Economics News 53:27 Vaccines For more updates, visit: http://www.brighteon.com/channel/hrreport NaturalNews videos would not be possible without you, as always we remain passionately dedicated to our mission of educating people all over the world on the subject of natural healing remedies and personal liberty (food freedom, medical freedom, the freedom of speech, etc.). Together, we're helping create a better world, with more honest food labeling, reduced chemical contamination, the avoidance of toxic heavy metals and vastly increased scientific transparency. ▶️ Every dollar you spend at the Health Ranger Store goes toward helping us achieve important science and content goals for humanity: https://www.healthrangerstore.com/ ▶️ Sign Up For Our Newsletter: https://www.naturalnews.com/Readerregistration.html ▶️ Brighteon: https://www.brighteon.com/channels/hrreport ▶️ Download our app: https://www.naturalnews.com/App ▶️ Join Our Social Network: https://brighteon.social/@HealthRanger ▶️ Check In Stock Products at: https://PrepWithMike.com

This MCAT podcast covers viruses. I cover virus structure, viral life cycle, transduction vs transfection, classification, viral mutation, and subviral particles. [01:17] Definition of a Virus [02:14] Virus Structure [11:15] Viral Life Cycle [18:02] Bacteriophage Life Cycles [21:41] Retrovirus Life Cycle [25:36] Difference Between Virus Transduction and Transfection [28:17] Virus Classification [33:33] Virus Mutation [41:52] Subviral Particles For comments and concerns, email me: MCATpodcast@medschoolcoach.com Thanks for listening!

The TWiV team explains how infectious horsepox virus - likely the ancestor of smallpox vaccines - was recovered from chemically synthesized DNA fragments. Hosts: Vincent Racaniello, Dickson Despommier, Alan Dove, Rich Condit, and Kathy Spindler Become a patron of TWiV! Links for this episode ASV 2018: asv.org, asv2018.umd.edu Potential Zika virus related birth defects, USA (MMWR) Bordering on an outbreak (Tex Obs) Horsepox virus from synthesized DNA (PLoS One) Horsepox based smallpox vaccine (NEJM) Has horsepox become extinct? (Vet Rec) Evolutionary path of smallpox vaccines (Lancet Inf Dis) Equination (Vaccine) Image credit Letters read on TWiV 478 Weekly Science Picks Kathy - UM EEB photo contest Rich - Dino-Lite digital microscope and digital microscope camera Dickson - Sandford Photography Alan - A list of female virologists Vincent - Virology Lectures 2018 Listener Picks Andrew - Interferome Neva - The Untreatable by Gavin Francis Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

David Gaughran is an Irish writer who spends most of his time travelling the world, collecting stories. He is the author of the Latin American historical adventures Mercenary and A Storm Hits Valparaiso, the short stories If You Go Into The Woods and Transfection, and the popular writers' guides Let's Get Digital and Let's Get Visible. He runs the publishing blog Let's Get Digital and the Latin American history site South Americana, and his work has been featured in the Huffington Post, The Sunday Times, and the Irish Times. I caught up with David at this year's Festival of Writing in York where he was giving several presentations about self-publishing.

David Gaughran is an Irish writer who spends most of his time travelling the world, collecting stories. He is the author of the Latin American historical adventures Mercenary and A Storm Hits Valparaiso, the short stories If You Go Into The Woods and Transfection, and the popular writers’ guides Let’s Get Digital and Let’s Get Visible. He runs the publishing blog Let’s Get Digital and the Latin American history site South Americana, and his work has been featured in the Huffington Post, The Sunday Times, and the Irish Times. I caught up with David at this year's Festival of Writing in York where he was giving several presentations about self-publishing. --- Send in a voice message: https://anchor.fm/self-publishing-journeys/message

The prognosis for cats with fibrosarcoma is still poor as the treatment options existing to date do not lead to satisfying results. After sole surgical removal of the tumor, up to 70 % of the cats develop local recurrences. Even with adjuvant radiation and/or chemotherapy, the recurrence rate can just be reduced to at most 40 %. In the present work an alternative treatment method in terms of neoadjuvant immunostimulatory gene therapy should be established. Via plasmids, three feline cytokine genes were transferred into the tumor. The plasmids coded for feIL-2, feIFN-γ and feGM-CSF. Using magnetofection, gene transfer should be optimized. The aim of this clinical dose escalation study was to define a well tolerated dose, which can be tested for its efficacy in a subsequent phase-II trial. Therefore the cats were examined at defined time points for treatment-related toxicity up to 180 days after surgery. Two more check-ups on day 270 and 360 after surgery were performed to elongate the observation period for local recurrences. Prerequisite for a cat’s admission to the study was the localization of the fibrosarcoma (primary tumor or recurrence) on the trunk. The tumor had to be excised in one setting without limb amputation. Affected cats were neither allowed to be pretreated with chemo-, radiation- or gene therapy nor with corticosteroids within the past six weeks. Further exclusion criteria were hints for metastases or other severe illnesses which reduce life expectancy to less than one year. Due to the potential teratogenic effect of the expressed cytokines, pregnancy had to be ruled out. Only cats with histopathologically confirmed fibrosarcoma continued the study after surgery. As this was a scientific study with a drug not registered yet, written informed consent from the owners was a prerequisite for the participation of each cat. Four treatment groups with defined dose escalation were prospectively fixed. The dose of the feline cytokines was 15, 50, 150 and 450 µg per plasmid in group I, II, III and IV. The initial dose (3 x 15 µg) was oriented to the total dose for small oral melanomas in dogs (400 µg) established by DOW et al. and is 1/10 of it. Plasmids as non-viral vectors were chosen for several reasons. Their handling is not liable to such strict regulations as the potentially more dangerous viral vectors. Their production is simple and affordable. They induce fewer side effects than viral vectors. Therefore plasmids are more suitable for application in veterinary clinical practice. Equal amounts of the plasmids were brought into 0.9 % saline. The positively charged magnetic nanoparticles were brought into solution with aqua for injections and were mixed with the plasmid formulation in a 1:1 ratio. This formulation had a total volume of 500 µl and was injected twice intratumorally in weekly intervals. Transfection was enhanced and targeted to the tumor area by the application of a neodymium-iron-boron magnet for the duration of 60 minutes. One week after the second application, wide en-bloc resection of the tumor was performed. Four cats were assigned to each treatment group. As questionable toxicity occured in group IV, four more cats were added to this group. A control group also consisting of four cats received surgery without neoadjuvant therapy. For ethical reasons, the application of empty plasmids was avoided so that in this group surgery could be performed without delay. Medical care of all the cats was carried out by the same team of internists, anesthetists and surgeons. Clinical and laboratory parameters were evaluated according to the VCOG-CTCAE system. All adverse events were registered, classified with severity grades and correlated to treatment. Plasma samples of all cats up to 14 days after surgery were examined for the existence of feGM-CSF and feIFN-γ with commercially available ELISA kits. Statistical analyses were performed comparing the treatment groups themselves as well as treatment groups and the control group regarding body weight, white blood cells and differential blood counts. Only one cat out of group IV showed adverse events during the neoadjuvant treatment period, which were classified as grade 3 and which were probably correlated to treatment (correlation grade 4). For this reason four more cats were treated with the highest dose. None of these cats showed side effects that could be correlate to treatment. The occurrence of early recurrences in four cats of group IV was outstanding, but of course, the expressiveness of this statement is low regarding the small group sizes. However it is known that biological drugs, especially IL-2, often do not have a linear dose-response profile. Therefore the optimal effective dose lies within a strictly defined area and is probably lower than the maximal tolerated dose. The highest applied dose which is 450 µg per plasmid was defined as a well tolerated dose. It can be safely tested for its efficacy in a subsequent phase-II trial. Because of the reflections mentioned above it would undoubtedly be convenient to test the third dose (150 µg per plasmid) in parallel for its effectiveness.

Ion channels are integral membrane proteins present in all cells. They are highly selective and assure a high rate for ions down their electrochemical gradient. In particular, small-conductance calcium-activated potassium channels (SK) are conducting potassium ions and are activated by binding of calcium ions to calmodulin, which is constitutively bound to the carboxy-terminus of each SK channel -subunit. Until now, only three SK channel subunits have been cloned, SK1, SK2 and SK3. Sequence alignment shows that the transmembrane and pore regions are highly conserved, while a high grade of divergence is observed in the amino- and carboxy-termini of the three subunits. In order to determine the expression of the different SK channel subtypes, pharmacological tols such as apamin and d-tubocurarine have been widely used. In this work, I show the characterization of a novel toxin, tamapin, isolated from the scorpion Mesobuths tamulus, which targets SK channels. Our experiments show that this toxin is more potent in blocking SK2 channels than apamin. Furthermore, tamapin only blocked the SK1 and SK3 channels at higher concentrations, with higher efficiency to block SK3 than SK1. Therefore, tamapin should be a good pharmacological tool to determine the molecular composition of native SK channels underlying calcium-activated potassium currents in various tissues. Secondly, I determined the molecular mechanism that prevents the formation of functional SK1 channels cloned from the rat brain (rSK1). Until now, little information was available on the rSK1 channels. rSK1 shows highly sequence identity (84%) with the human homologue, hSK1. hSK1 subunits form functional potassium channels that are blocked by apamin and d-tubocurarine. However, when I expressed rSK1 in HEK-293 cells no potassium currents above background were observed, although immunofluorescence experiments using a specific antibody against the rSK1 protein showed expression of the channel. I generated rSK1 core chimeras in which I exchanged the amino-and/carboxy-terminus with the same region of rSK2 or hSK1. Exchange of amino-and carboxy-terminus or only of the carboxy-terminus resulted in the formation of functional potassium channels. Furthermore, I used these functional chimeras to determine the toxin sensitivity of rSK1 for apamin and d-tubocurarine. Surprisingly, when these blockers wre applied, no sensitivity was observed, although hSK1 and rSK1 show a complete sequence identity in the pore region, which is suggested to contain the binding site for apamin. Finally, I characterized a novel splice variant of the calcium-activated potassium channel subunit rSK2, referred to as rSK2-860. The rSK2-860 cDNA codes for a protein which is 275 amino acids longer at the amino-terminus when compared with originally cloned rSK2 subunit. Transfection of rSK2-860 in different cell lines resulted in a surprising expression pattern of the protein. Th protein formed small clusters around the cell nucleus, but no membrane stain could be observed. This data shows that the additional 275 amino acid-long stretch at the amino-terminus is responsible for retention and clustering of rSK2-860 protein. In order to narrow down the region responsible for this phenotype, I generated truncated proteins. This resulted in the isolation of an 100 amino acid-long region that seems to be responsible for the retention and clustering of rSK2-860 channels. Further truncations and deletions could help us to find the exact signal which is responsible for this characteristic behavior of the rSK2-860 protein.

The regulation of murine cytomegalovirus early (E) gene expression was studied in the cell line B25, which is stably transfected with the immediate-early ie1/ie3 gene complex. Infection of B25 cells in the presence of the protein synthesis inhibitor cycloheximide resulted in the expression of some E genes, whereas for the expression of other E genes prior protein synthesis was still mandatory, thus showing differences in the expression requirements of individual E genes. Transcription unit e1, a member of the E genes induced by immediate-early products of the ie1/ie3 gene complex, was characterized. It is located between map units 0.709 and 0.721 of the genome of murine cytomegalovirus strain Smith. A 2.6-kilobase RNA specified in this region is spliced from three exons of 912, 177, and 1,007 or 1,020 nucleotides, which are separated by introns of 93 and 326 nucleotides. The second AUG located in the first exon 119 nucleotides downstream of the 5' cap site is followed by an open reading frame of 990 nucleotides. The predicted polypeptide of 330 amino acids has a calculated molecular mass of 36.4 kilodaltons. Transfection with e1 revealed three antigenically related proteins of 36, 37, and 38 kilodaltons; these proteins probably represent differently modified forms of the predicted protein. These three proteins are phosphorylated and are associated with intranuclear inclusion bodies. A 33-kilodalton protein also derived from e1 was identified as a product of nonspliced transcripts. Comparison of amino acid sequences revealed homology between the murine cytomegalovirus transcription unit e1 and a human cytomegalovirus E transcription unit.

Sun, 1 Jan 1989 12:00:00 +0100 http://epub.ub.uni-muenchen.de/3071/ http://epub.ub.uni-muenchen.de/3071/1/3071.pdf Thureau, S. R.; Wildner, G.; Kuon, W.; Weiß, Elisabeth; Riethmüller, Gert Thureau, S. R.; Wildner, G.; Kuon, W.; Weiß, Elisabeth und Riethmüller, Gert (1989): Expression and immmunogenicity of HLA-B27 in high-transfection recipient P815:. a new method to induce monoclonal antibodies directed against HLA-B27. In: Tissue Antigens, Vol. 33: pp. 511-519. B