Podcasts about gsk3

BUFFALO, NY - April 11, 2025 – A new research perspective was published in Oncotarget, Volume 16, on April 4, 2025, titled “GSK3β activation is a key driver of resistance to Raf inhibition in BRAF mutant melanoma cells." In this work, first author Diana Crisan and corresponding author Abhijit Basu from the University Hospital Ulm led a team that presents experimental evidence pointing to the protein GSK3β as a key contributor to drug resistance in melanoma. Their findings suggest that GSK3β becomes increasingly active in cancer cells during treatment, helping them survive and adapt despite ongoing therapy with BRAF inhibitors. Melanoma is a type of skin cancer in which nearly half of patients have mutations in the BRAF gene that accelerate tumor growth. While treatments targeting BRAF, known as BRAF inhibitors, initially work well, tumors often find ways to fight back. This research perspective explores how GSK3β, a protein involved in metabolism and cell survival, becomes more active in melanoma cells that develop resistance to BRAF inhibitors. Researchers treated melanoma cells with a common BRAF mutation using Dabrafenib, a widely used BRAF inhibitor. Over time, the cancer cells developed resistance and showed a marked increase in GSK3β levels. This pattern was confirmed across multiple melanoma cell models, suggesting that the finding is consistent and reliable. Importantly, the researchers observed that treating resistant cancer cells with a GSK3β inhibitor significantly reduced their growth. This result suggests that blocking this protein could restore sensitivity to treatment, highlighting GSK3β as a promising therapeutic target and supporting the idea of combining GSK3β inhibitors with existing melanoma therapies. “Remarkably, treatment of BRAFi-resistant melanoma cells with the GSK3 inhibitor LY2090314 for three weeks could overcome resistance and significantly decreased melanoma cell growth, confirming the causal role of GSK3 activation for BRAFi resistance development.” The research perspective adds to ongoing efforts to understand and overcome melanoma drug resistance. It shows that resistance is not driven only by genetic mutations but may also involve adaptive changes in the cell's internal signaling and survival mechanisms. By identifying GSK3β as a potential contributor, the authors offer a new direction for improving the durability of targeted treatments in melanoma. As research continues, GSK3β may be a critical factor in the long-term success of melanoma therapy, particularly for patients who have stopped responding to standard BRAF-targeted drugs. Continue reading: DOI: https://doi.org/10.18632/oncotarget.28711 Correspondence to: Abhijit Basu — abhijit.basu@alumni.uni-ulm.de Video short - https://www.youtube.com/watch?v=G2Tq4_r6xLw Subscribe for free publication alerts from Oncotarget - https://www.oncotarget.com/subscribe/ About Oncotarget Oncotarget (a primarily oncology-focused, peer-reviewed, open access journal) aims to maximize research impact through insightful peer-review; eliminate borders between specialties by linking different fields of oncology, cancer research and biomedical sciences; and foster application of basic and clinical science. Oncotarget is indexed and archived by PubMed/Medline, PubMed Central, Scopus, EMBASE, META (Chan Zuckerberg Initiative) (2018-2022), and Dimensions (Digital Science). To learn more about Oncotarget, please visit https://www.oncotarget.com and connect with us: Facebook - https://www.facebook.com/Oncotarget/ X - https://twitter.com/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/@OncotargetJournal LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/ Spotify - https://open.spotify.com/show/0gRwT6BqYWJzxzmjPJwtVh MEDIA@IMPACTJOURNALS.COM

Thursday, September 26th, 2024Today, the Jack Smith 180 page immunity briefing is due today; every Teamsters union in Florida has endorsed Kamala Harris; a Wisconsin mayor is caught stealing a ballot drop box; Alabama Republicans are working to block Glock switches which are like bump stocks for handguns; Georgia Democrats are suing Brian Kemp to force an ethics investigation into the State Elections Board; the House passes a clean government funding bill and are now on break until after the election; Hurricane Helene will jump to a category 4 before landfall in Florida; a top Muslim organization has endorsed Kamala Harris; and the Supreme Court refuses to stay the execution of Marcellus Williams; and Allison delivers your Good News. For a limited time, HomeChef is offering you 18 Free Meals, plus Free Shipping on your first box, and Free Dessert for Life. At https://www.HomeChef.com/DAILYBEANS.Join AG At ‘Creatives for Harris' Virtual Rally 9.26.24 9PM ET, 6PM PT Registration LinkHarris Campaign Social Media Toolkit (kamalaharris.com)Give to the Kamala Harris Presidential CampaignKamala Harris — Donate via ActBlue (MSW Media's Donation Link)Come See AG Saturday, September 28th At The Sexy Liberal Save The World Comedy Tour!Phoenix, Arizona - Get Tickets at: https://sexyliberal.comGuests:Generation Data Co-Founders Nancy Stalnaker (Data Diva) and Matt Mawhinneygenerationdata.orgA Special Excel Training For Daily Beans Listeners!Saturday, October 12 · 10am - 1pm PDTgenerationdata.org/daily-beansStoriesWisconsin mayor carts away absentee ballot drop box, says he did nothing wrong (CNN)Democrats sue over Georgia rules they say could block election certifications (CBS News)GOP lawmakers now back Glock switch ban after mass shooting at Birmingham Hush lounge (AL.com)Every Teamsters union in Florida backs Kamala Harris for President (Florida Politics)Top Muslim-voter organization endorses Harris as Middle East conflict escalates (AP News)Missouri executes Marcellus Williams despite questions over evidence, after Supreme Court denies final bid for delay (CBS News) From The Good NewsWorkforce Innovation and Opportunity Act (dot.gov)The Veterans Access, Choice And Accountability Act Of 2014 (house.gov)The Charlatans - Weirdo HD (YouTube)Inhibition of GSK3α,β rescues cognitive phenotypes in a preclinical mouse model of CTNNB1 syndrome (embopress.org)Researchers Identify Possible Treatment for Rare Disorder (tufts.edu)She's Fly focuses on quality outdoor gear made for women, by women (shesfly.com)So What Else (Food Bank And More | DC Area | sowhatelse.org)If any DC area listeners can help, please email info@sowhatelse.orgCheck Your Voter Registration!vote.orgThere is a new “Harris For President” Patreon tier:https://www.patreon.com/muellershewrote/membership Check out other MSW Media podcastshttps://mswmedia.com/shows/Subscribe for free to MuellerSheWrote on Substackhttps://muellershewrote.substack.com Follow AG and Dana on Social MediaDr. Allison Gill https://muellershewrote.substack.comhttps://twitter.com/MuellerSheWrotehttps://www.threads.net/@muellershewrotehttps://www.tiktok.com/@muellershewrotehttps://instagram.com/muellershewroteDana Goldberghttps://twitter.com/DGComedyhttps://www.instagram.com/dgcomedyhttps://www.facebook.com/dgcomedyhttps://danagoldberg.comHave some good news; a confession; or a correction to share?Good News & Confessions - The Daily Beanshttps://www.dailybeanspod.com/confessional/ Listener Survey:http://survey.podtrac.com/start-survey.aspx?pubid=BffJOlI7qQcF&ver=shortFollow the Podcast on Apple:The Daily Beans on Apple PodcastsWant to support the show and get it ad-free and early?Supercasthttps://dailybeans.supercast.com/OrPatreon https://patreon.com/thedailybeansOr subscribe on Apple Podcasts with our affiliate linkThe Daily Beans on Apple Podcasts

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.26.538498v1?rss=1 Authors: Li, E., Benitez, C., Boggess, S. C., Koontz, M., Rose, I. V. L., Draeger, N., Teter, O. M., Samelson, A. J., Ullian, E. M., Kampmann, M. Abstract: The sheer complexity of the brain has complicated our ability to understand its cellular mechanisms in health and disease. Genome-wide association studies have uncovered genetic variants associated with specific neurological phenotypes and diseases. In addition, single-cell transcriptomics have provided molecular descriptions of specific brain cell types and the changes they undergo during disease. Although these approaches provide a giant leap forward towards understanding how genetic variation can lead to functional changes in the brain, they do not establish molecular mechanisms. To address this need, we developed a 3D co-culture system termed iAssembloids (induced multi-lineage assembloids) that enables the rapid generation of homogenous neuron-glia spheroids. We characterize these iAssembloids with immunohistochemistry and single-cell transcriptomics and combine them with large-scale CRISPRi-based screens. In our first application, we ask how glial and neuronal cells interact to control neuronal death and survival. Our CRISPRi-based screens identified that GSK3{beta} inhibits the protective NRF2-mediated oxidative stress response in the presence of reactive oxygen species elicited by high neuronal activity, which was not previously found in 2D monoculture neuron screens. We also apply the platform to investigate the role of APOE-{epsilon}4, a risk variant for Alzheimer's Disease, in its effect on neuronal survival. This platform expands the toolbox for the unbiased identification of mechanisms of cell-cell interactions in brain health and disease. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.13.536716v1?rss=1 Authors: Darmasaputra, G., Chuva de Sousa Lopes, S. M., Clevers, H. C., Galli, M. Abstract: Many plant and animal cells transition from canonical to non-canonical cell cycles during development, resulting in the formation of polyploid cells. Two types of non-canonical cell cycles exist: endoreplication, where cells increase their DNA content without entering M phase, and endomitosis, where cells enter M phase but exit prematurely. Although endoreplication has been extensively studied in plants and insects, much less is known on the regulation of endomitosis, which is the most common mode of polyploidization in mammals. In this study, we use fetal-derived human hepatocyte organoids (Hep-Org), to investigate how human hepatocytes initiate and execute endomitosis. We find that cells in endomitosis M phase have normal mitotic timings, but lose membrane anchorage to the midbody during cytokinesis, resulting in regression of the cytokinetic furrow and formation of binucleate cells. Using immunofluorescence, we find that three cortical anchoring proteins, RacGAP1, anillin, and citron kinase (CIT-K), lose their association with the cell cortex during cytokinetic regression. Moreover, reduction of WNT activity by withdrawal of CHIR99021, a GSK3 inhibitor, from the culturing medium increases the percentage of binucleated cells in Hep-Orgs. This effect is lost in organoids with mutations in the atypical E2F proteins, E2F7 and E2F8, which have been implicated in binucleation of rodent hepatocytes. Together, our results identify how human hepatocytes inhibit cell division in endomitosis, and highlight an evolutionary recurrent mechanism to initiate non-canonical cell cycles in mammals. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.11.536245v1?rss=1 Authors: Takla, T. N., Luo, J., Sudyk, R., Huang, J., Walker, J. C., Vora, N. L., Sexton, J. Z., Parent, J. M., Tidball, A. M. Abstract: Neural tube defects (NTDs) including anencephaly and spina bifida are common major malformations of fetal development resulting from incomplete closure of the neural tube. These conditions lead to either universal death (anencephaly) or life-long severe complications (spina bifida). Despite hundreds of genetic mouse models having neural tube defect phenotypes, the genetics of human NTDs are poorly understood. Furthermore, pharmaceuticals such as antiseizure medications have been found clinically to increase the risk of NTDs when administered during pregnancy. Therefore, a model that recapitulates human neurodevelopment would be of immense benefit to understand the genetics underlying NTDs and identify teratogenic mechanisms. Using our self-organizing single rosette spheroid (SOSRS) brain organoid system, we have developed a high-throughput image analysis pipeline for evaluating SOSRS structure for NTD-like phenotypes. Similar to small molecule inhibition of apical constriction, the antiseizure medication valproic acid (VPA), a known cause of NTDs, increases the apical lumen size and apical cell surface area in a dose-responsive manner. This expansion was mimicked by GSK3-{beta} and HDAC inhibitors; however, RNA sequencing suggests VPA does not inhibit GSK3-{beta} at these concentrations. Knockout of SHROOM3, a well-known NTD-related gene, also caused expansion of the lumen as well as reduced f-actin polarization. The increased lumen sizes were caused by reduced cell apical constriction suggesting that impingement of this process is a shared mechanism for VPA treatment and SHROOM3-KO, two well-known causes of NTDs. Our system allows the rapid identification of NTD-like phenotypes for both compounds and genetic variants and should prove useful for understanding specific NTD mechanisms and predicting drug teratogenicity. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.03.18.533245v1?rss=1 Authors: Sarkar, S., Gharami, K., Paidi, R. K., Srikumar, B. N., Biswas, S. C. Abstract: Astrocytes respond to any pathological insult to brain including Alzheimer's disease (AD) through 'reactive astrogliosis'. Recently, we demonstrated that tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), a neuroprotective cytokine, is released through reactive astrogliosis early in response to amyloid-{beta} (A{beta}). Here, we show that TIMP-1 has superior mechanism-of-actions on neurons in models of AD. It not only confers neuroprotection against A{beta}-induced apoptosis and autophagy via CD63 receptor but also displays synapse-specific effects that underlie cognitive recovery in AD. We detect diminished levels of TIMP-1 in the hippocampi of 5xFAD mice versus wild-type counterparts. Interestingly, exogenous TIMP-1 injection in this transgenic model ameliorates its cognitive functions, likely by restoring long-term potentiation at hippocampal synapses. We observe BDNF induction and GSK3{beta} inhibition, which may be the key TIMP-1-driven underlying mechanisms at synapses. Thus, we show an astrocyte-origin cytokine-driven mechanism for synaptic and cognitive salvation promising an exciting avenue in AD therapeutic research. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.14.528444v1?rss=1 Authors: Orrico-Sanchez, A., Guiard, B., Manta, S., Callebert, J., Launay, J.-M., Louis, F., Paccard, A., Gruszczynski, C., Betancur, C., Vialou, V., Gautron, S. L. Abstract: Selective serotonin reuptake inhibitors (SSRI) are common first-line treatments for major depression. However, a significant number of depressed patients do not respond adequately to these pharmacological treatments. In the present preclinical study, we demonstrate that organic cation transporter 2 (OCT2), an atypical monoamine transporter, contributes to the effects of SSRI by regulating the routing of the essential amino acid tryptophan to the brain. Contrarily to wild-type mice, OCT2-invalidated mice failed to respond to prolonged fluoxetine treatment in a chronic depression model induced by corticosterone exposure recapitulating core symptoms of depression, i.e., anhedonia, social withdrawal, anxiety and memory impairment. Levels of tryptophan and its metabolites serotonin and kynurenine were decreased in the brain of OCT2 mutant mice compared to wild-type mice and reciprocally increased in mutants' plasma, at basal state and after fluoxetine treatment. OCT2 was detected by immunofluorescence in several structures at the blood-cerebrospinal fluid (CSF) or brain-CSF interface. Tryptophan supplementation during fluoxetine treatment increased brain concentrations of tryptophan and serotonin in wild-type and OCT2 mutant mice, yet more efficiently in WT than in mutants. Importantly, tryptophan supplementation improved the sensitivity to fluoxetine treatment of OCT2 mutant mice, impacting chiefly anhedonia and short-term memory. Western blot analysis showed that glycogen synthase kinase-3{beta} (GSK3{beta}) and mammalian/mechanistic target of rapamycin (mTOR) intracellular signaling was impaired in OCT2 mutant mice brain after corticosterone and fluoxetine treatment and, conversely, tryptophan supplementation recruited selectively the mTOR protein complex 2. This study provides the first evidence of the physiological relevance of OCT2-mediated tryptophan transport, and its biological consequences on serotonin homeostasis in the brain and SSRI efficacy. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.09.527935v1?rss=1 Authors: Ong, H. W., Liang, Y., Richardson, W., Lowry, E. R., Wells, C. I., Chen, X., Silvestre, M., Dempster, K., Silvaroli, J. A., Smith, J. L., Wichterle, H., Pabla, N. S., Ultanir, S. K., Bullock, A. N., Drewry, D., Axtman, A. D. Abstract: Despite mediating several essential processes in the brain, including during development, cyclin-dependent kinase-like 5 (CDKL5) remains a poorly characterized human protein kinase. Accordingly, its substrates, functions, and regulatory mechanisms have not been fully described. We realized that availability of a potent and selective small molecule probe targeting CDKL5 could enable illumination of its roles in normal development as well as in diseases where it has become aberrant due to mutation. We prepared analogs of AT-7519, a known inhibitor of several cyclin dependent and cyclin-dependent kinase-like kinases that has been advanced into Phase II clinical trials. We identified analog 2 as a highly potent and cell-active chemical probe for CDKL5/GSK3 (glycogen synthase kinase 3). Evaluation of its kinome-wide selectivity confirmed that analog 2 demonstrates excellent selectivity and only retains GSK3/{beta} affinity. As confirmation that our chemical probe is a high-quality tool to use in directed biological studies, we demonstrated inhibition of downstream CDKL5 and GSK3/{beta} signaling and solved a co-crystal structure of analog 2 bound to CDKL5. A structurally similar analog (4) proved to lack CDKL5 affinity and maintain potent and selective inhibition of GSK3/{beta}. Finally, we used our chemical probe pair (2 and 4) to demonstrate that inhibition of CDKL5 and/or GSK3/{beta} promotes the survival of human motor neurons exposed to endoplasmic reticulum (ER) stress. We have demonstrated a neuroprotective phenotype elicited by our chemical probe pair and exemplified the utility of our compounds to characterize the role of CDKL5/GSK3 in neurons and beyond. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.01.26.524308v1?rss=1 Authors: Chan, V., Bone, L., Anderson, K. E., Zhang, K., Orofiamma, L., Awadeh, Y., Lee, D. K. C., Fu, N. J., Chow, J. T. S., Salmena, L., Stephens, L. R., Hawkins, P. T., Antonescu, C. N., Botelho, R. J. Abstract: Receptor tyrosine kinases such as epidermal growth factor (EGF) receptor (EGFR) stimulate phosphatidylinositol 3-kinases (PI3Ks) to convert phosphaitydlinositol-4,5-bisphosophate [PtdIns(4,5)P2] into phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3]. PtdIns(3,4,5)P3 then promotes various pathways leading to actin remodelling, changes in gene expression, and enhanced anabolic activity, cell survival and proliferation. In part, PtdIns(3,4,5)P3 achieves these functions by stimulating the kinase Akt, which phosphorylates numerous targets like Tsc2 and GSK3{beta}. Overall, unchecked upregulation of PtdIns(3,4,5)P3-Akt signalling can promote tumourgenesis and cancer progression. Interestingly, 50-70% of PtdIns and PtdInsPs have stearate and arachidonate at sn-1 and sn-2 positions of glycerol, respectively, forming a species known as 38:4-PtdIns/PtdInsPs. It is thought that LCLAT1/LYCAT and MBOAT7/LPIAT1 acyltransferases are respectively responsible for enriching PtdIns with this acyl composition. We previously showed that disruption of LCLAT1 altered the acyl profile of bis-phosphorylated PtdInsPs, lowered PtdIns(4,5)P2, and perturbed endocytosis and endocytic trafficking. However, the role of LCLAT1 in receptor tyrosine kinase and PtdIns(3,4,5)P3 signaling was not explored. Here, we show that LCLAT1 silencing in MDA-MB-231 and ARPE-19 cells abated the levels of PtdIns(3,4,5)P3 in response to EGF signalling. Importantly, LCLAT1-silenced cells were also impaired for EGF-mediated Akt activation and downstream signalling, and consequently, were depressed for cell proliferation and survival. Thus, our work provides first evidence that the LCLAT1 acyltransferase supports receptor tyrosine kinase signalling through the PtdIns(3,4,5)P3-Akt axis and may represent a novel target for therapeutic development against cancers. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.01.12.523372v1?rss=1 Authors: Mendez-Vazquez, H., Roach, R. L., Nip, K., Sathler, M. F., Garver, T., Danzman, R. A., Moseley, M. C., Roberts, J. P., Koch, O. N., Steger, A. A., Lee, R., Arikkath, J., Kim, S. Abstract: {delta}-catenin is expressed in excitatory synapses and functions as an anchor for the glutamatergic AMPA receptor (AMPAR) GluA2 subunit in the postsynaptic density. The glycine 34 to serine (G34S) mutation in the{delta} -catenin gene is found in autism spectrum disorder (ASD) patients and induces loss of {delta}-catenin functions at excitatory synapses, which is presumed to underlie ASD pathogenesis in humans. However, how the G34S mutation causes loss of {delta}-catenin functions to induce ASD remains unclear. Here, using neuroblastoma cells, we discover that the G34S mutation generates an additional phosphorylation site for glycogen synthase kinase 3{beta} (GSK3{beta}). This promotes {delta}-catenin degradation and causes the reduction of {delta}-catenin levels, which likely contributes to the loss of {delta}-catenin functions. Synaptic {delta}-catenin and GluA2 levels in the cortex are significantly decreased in mice harboring the {delta}-catenin G34S mutation. The G34S mutation increases glutamatergic activity in cortical excitatory neurons while it is decreased in inhibitory interneurons, indicating changes in cellular excitation and inhibition. {delta}-catenin G34S mutant mice also exhibit social dysfunction, a common feature of ASD. Most importantly, inhibition of GSK3{beta} activity reverses the G34S-induced loss of {delta}-catenin function effects in cells and mice. Finally, using {delta}-catenin knockout mice, we confirm that {delta}-catenin is required for GSK3{beta} inhibition-induced restoration of normal social behaviors in {delta}-catenin G34S mutant animals. Taken together, we reveal that the loss of {delta}-catenin functions arising from the ASD-associated G34S mutation induces social dysfunction via alterations in glutamatergic activity and that GSK3{beta} inhibition can reverse {delta}-catenin G34S-induced synaptic and behavioral deficits. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.01.10.523252v1?rss=1 Authors: del Puerto, A., Marti-Prado, B., Barrios-Munoz, A. L., Lopez-Fonseca, C., Pose-Utrilla, J., Alcover-Sanchez, B., Cesca, F., Schiavo, G., Campanero, M. R., Farinas, I., Iglesias, T., Porlan, E. Abstract: In the adult mammalian brain, neural stem cells (NSCs) located in highly restricted niches sustain the generation of new neurons that integrate into existing circuits. A reduction in adult neurogenesis is linked to ageing and neurodegeneration, whereas dysregulation of proliferation and survival of NSCs have been hypothesized to be at the origin of glioma. Thus, unravelling the molecular underpinnings of the regulated activation that NSCs must undergo to proliferate and generate new progeny is of considerable relevance. current research has identified cues promoting or restraining NSCs activation. Yet, whether NSCs depend on external signals to survive or if intrinsic factors establish a threshold for sustaining their viability remains elusive, even if this knowledge could involve potential for devising novel therapeutic strategies. Kidins220 (Kinase D-interacting substrate of 220 kDa) is an essential effector of crucial pathways for neuronal survival and differentiation. It is dramatically altered in cancer and in neurological and neurodegenerative disorders, emerging as a regulatory molecule with important functions in human disease. Herein, we discover severe neurogenic deficits and hippocampal-based spatial memory defects in Kidins220 deficient mice. Mechanistically, we demonstrate that Kidins220-dependent activation of AKT in response to EGF restraints GSK3 activity preventing NSCs apoptosis. Hence, Kidins220 levels set a molecular threshold for survival in response to mitogens, allowing adult NSCs to proliferate. Our study identifies Kidins220 as a key player for sensing the availability of growth factors to sustain adult neurogenesis, uncovering a molecular link that may help paving the way towards neurorepair. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.12.14.520376v1?rss=1 Authors: Zhao, H., Chen, J., Qi, C., Wang, T., Liang, T., Hao, X., Li, X., Yin, X., Yu, T., Zhang, Y. Abstract: Restoring the normal structure and function of injured tendons is one of the biggest challenges faced by the Department of Orthopedics and Sports Medicine. Tendon-derived stem cells (TDSCs), a new type of pluripotent stem cells with multidirectional differentiation potential, are expected to be promising cell seeds for the treatment of tendon injury and tendon-bone healing in the future. In this study, tendon stem cells were successfully isolated from human tissues, which were positive for markers CD44, CD90, and CD105, and exhibited clonality and multilineage differentiation ability. Analysis of single-cell sequencing results and mass spectrometry identification results showed that there were differences in protein expression during CTGF-induced TDSC tendon differentiation. Reverse Co-IP, qPCR, WB, and immunofluorescence detection all confirmed that CTGF directly interacts with KIT, thereby mediating the transcription factor HES1 to regulate the Wnt/{beta}-catenin signaling pathway (GSK3{beta}, {beta}-catenin, TCF4). ChIP-qPCR and dual-luciferase reporter gene assays indicated that HES1 regulates stem cell differentiation by directly regulating the expression of GSK3{beta} in the Wnt/{beta}-catenin pathway. Rats were treated with TDSCs overexpressing the KIT gene after repair surgery. This method had a more ideal recovery effect than other methods through animal behavioral scores, mechanical properties testing, and HE staining tissue observation. This study found that the use of modified human tendon stem cells (hTDSC) could promote graft ligamentization and tendon-bone healing after ACL reconstruction, which could provide an effective way for faster and better recovery from tendon injury. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.12.09.519813v1?rss=1 Authors: Zhang, K., Da Silva, F., Seidl, C., Wilsch-Bräuninger, M., Herbst, J., Huttner, W. B., Niehrs, C. Abstract: WNT signalling is of paramount importance in development, stem cell maintenance, and disease. WNT ligands typically signal via receptor activation at the plasma membrane to induce {beta}-catenin-dependent gene activation. Here we show that in primary cilia, WNT receptors relay a WNT/GSK3 signal that {beta}-catenin-independently promotes ciliogenesis. Innovations supporting this conclusion are monitoring acute WNT co-receptor activation (phospho-LRP6) and identifying and mutating the LRP6 ciliary targeting sequence. Ciliary WNT signalling inhibits protein phosphatase 1 (PP1) activity, a negative regulator of ciliogenesis, by decommissioning GSK3-mediated phosphorylation of the PP1 regulatory inhibitor subunit PPP1R2. Accordingly, deficiency of WNT/GSK3 signalling by depletion of cyclin Y and cyclin-Y-like protein 1 induces widespread primary cilia defects in mouse embryonic neuronal precursors, kidney proximal tubules, and adult mice preadipocytes. We conclude that primary cilia are WNT PP1 signalling organelles. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.25.517972v1?rss=1 Authors: Maurizi, E., Merra, A., Macaluso, C., Schiroli, D., Pellegrini, G. Abstract: Human corneal endothelial cells are organized in a tight mosaic of hexagonal cells and serve a critical function in maintaining corneal hydration and clear vision. Regeneration of the corneal endothelial tissue is hampered by its poor proliferative capacity, which is partially retrieved in vitro, albeit only for a limited number of passages before the cells undergo mesenchymal transition (EnMT). Although different culture conditions have been proposed in order to delay this process and prolong the number of cell passages, EnMT has still not been fully understood and successfully counteracted. In this perspective, we identified herein a single GSK3 inhibitor, CHIR99021, able to revert and avoid EnMT in primary human corneal endothelial cells (HCEnCs) from old donors until late passages in vitro (P8), as shown from cell morphology analysis (circularity). In accordance, CHIR99021 reduced expression of alpha-SMA, an EnMT marker, while restored endothelial markers such as ZO-1, Na+/K+ ATPase and N-cadherin, without increasing cell proliferation. A further analysis on RNA expression confirmed CHIR99021 induced downregulation of EnMT markers (alpha-SMA and CD44), upregulation of the proliferation repressor p21 and revealed novel insights into the beta-catenin and TGFbeta; pathways intersections in HCEnCs. The use of CHIR99021 sheds light on the mechanisms involved in EnMT and brings a substantial advantage in maintaining primary HCEnCs in culture until late passages, while preserving the correct morphology and phenotype. Altogether, these results bring crucial advancements towards the improvement of the corneal endothelial cells based therapy. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.10.23.513322v1?rss=1 Authors: Chen, X., Rong, K., Han, W., Pang, Y., Chai, G. Abstract: In previous research, miR-148a-3p deficiency was observed in bone malformation in hemifacial microsomia. Herein, in this article, we probed into the role of miR-148a-3p in bone physiology by utilizing miR-148a knock-out (KO) mice. Compared with wild-type (WT) or heterozygotic (HE) littermates, miR-148a knock-out mice manifested lower body weight, bone dysplasia with increased bone mass. Through in-vitro experiments, in terms of miR-148a-3p overexpression (miRNA mimic transfection) and knockout (primary cells from WT and KO littermates), we found that miR-148a-3p can suppress osteogenesis, either in the ALP activity or bone nodules formation. Afterward, by means of proteomics, combined with RNA-sequencing and prediction databases of microRNA targets (miRDB and TargetScan), nine candidate genes targeted by miR-148a-3p were identified. Among them, only Itga11 was regulated by mRNA degradation, while the others were modulated via post-transcriptional inhibition. Based on several online databases (GenePaint, BioGPS, STRING), Integrin Subunit Alpha 11 (Itga11) was suggested to play an essential role in osteogenesis and it was confirmed as one direct target of miR-148a-3p by dual-luciferase reporter assay. Meanwhile, gene set enrichment analysis (GSEA) indicated activation of PI3K-Akt signaling pathway and WNT signaling pathway in miR-148a KO mice. The thereafter western blot confirmed that PI3K/Akt/GSK3/{beta}-catenin signaling pathway was involved. Taken together, we demonstrated that miR-148a-3p can inhibit osteogenesis by targeting Itga11 via PI3K/Akt/GSK3/{beta}-catenin pathway. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

When stressed, people are quicker to jump to the worst conclusion University College London, July 29, 2021 When under stress, people reach undesirable conclusions based on weaker evidence than when they are relaxed, finds a new study led by UCL researchers. The findings, published today in the Journal of Neuroscience, show that stress can make people more likely to conclude the worst scenario is true. Senior author Professor Tali Sharot (UCL Psychology & Language Sciences and Max Planck UCL Centre for Computational Psychiatry and Ageing Research) said: "Many of the most significant choices you will make, from financial decisions to medical and professional ones, will happen while you feel stressed. Often these decisions require you to first gather information and weigh the evidence. For example, you may consult multiple physicians before deciding on a best course of medical treatment. We wanted to find out: does feeling stressed change how you process and use the information you gather? "Our research suggests that under stress, people weight each piece of evidence that supports undesirable conclusions more than when they are relaxed. In contrast, how they weigh evidence that supports desirable conclusions is not affected by stress. As a result, people are more likely to conclude the worst is true when they are stressed." For the study, 91 volunteers played a categorisation game, in which they could gather as much evidence as they wanted to decide whether they were in a desirable environment (which was associated with rewards) or an undesirable environment (which was associated with losses). They were incentivised for accuracy. Prior to playing the game, 40 of the volunteers were told that they had to give a surprise public speech, which would be judged by a panel of experts. This caused them to feel stressed and anxious. The researchers found that under stress, the volunteers needed weaker evidence to reach the conclusion that they were in the undesirable environment. By contrast, stress did not change the strength of the evidence needed to reach the conclusion that they were in the desirable environment. Lead author, PhD student Laura Globig (UCL Psychology & Language Sciences and Max Planck UCL Centre for Computational Psychiatry and Ageing Research) said that "we usually think of stressful situations as a hindrance to our decision-making process. But the pattern of learning we have uncovered may counterintuitively be adaptive, because negative beliefs may drive people to be extra cautious when in threatening environments."   Thai Cabinet Approves Use of Fah Talai Jone (green chiretta) to Treat Asymptomatic COVID-19 Cases Bangkok Post, July 22, 2021 Prime Minister Prayut Chan-o-cha has instructed authorities to set up a committee to study the use of green chiretta (Andrographis paniculata) extracts to treat Covid-19 patients with mild symptoms. The announcement was made at a cabinet meeting on Tuesday, which was convened to discuss additional measures to help curb the Covid-19 outbreak, the premier said on Wednesday. Deputy Prime Minister and Public Health Minister Anutin Charnvirakul was appointed head of the committee. It will coordinate studies on the safety and efficacy of green chiretta extracts on Covid-19 patients, as well as draft a strategic plan to promote Thai traditional medicine in general. The decision was taken in response to a proposal from Justice Minister Somsak Thepsutin, who urged the government to scale up the use of traditional medicines on Covid-19 patients with mild symptoms. This comes amid a vaccine shortage which has led to criticism for the government. His proposal came with evidence showing the Department of Corrections' success in treating 12,376 inmates who were infected with Covid-19 with green chiretta extracts. Of this number, 5,045 inmates were in Chiang Mai Central Prison, 2,100 in Nonthaburi Provincial Prison and 5,231 in Bang Kwang Central Prison also in Nonthaburi, said Mr Somsak. Before prescribing the herbal medicine to infected inmates, Mr Somsak said he had studied information by the Department of Thai Traditional and Alternative Medicine, which recommended a dosage of 180mg of andrographolides from green chiretta for five consecutive days to patients with mild symptoms. Citing the same research, the minister said each rai of land can yield up to 600kg of green chirettas, which can be turned into roughly 375,000 herbal extract capsules, he said. A total of 3.1 billion such capsules will be needed to cover all Thais, which means 8,400 rai of land will need to be planted with the herb, he said. The Department of Corrections now plans to produce about 50 million capsules of the herbal medicine in the next four months, which it aims to prescribe to about 50% of the prison population, he said. Due to its medical benefits, green chiretta has become a cash crop which is now in high demand in the export sector, he said. Mr Somsak added that the medicinal herb costs about 450 baht per kg    Taking breaks while learning improves memory   Max Planck Institute of Neurobiology (Germany), July 29, 2021 We remember things longer if we take breaks during learning, referred to as the spacing effect. Scientists at the Max Planck Institute of Neurobiology gained deeper insight into the neuronal basis for this phenomenon in mice. With longer intervals between learning repetitions, mice reuse more of the same neurons as before—instead of activating different ones. Possibly, this allows the neuronal connections to strengthen with each learning event, such that knowledge is stored for a longer time. Many of us have experienced the following: the day before an exam, we try to cram a huge amount of information into our brain. But just as quickly as we acquired it, the knowledge we have painstakingly gained is gone again. The good news is that we can counteract this forgetting. With expanded time intervals between individual learning events, we retain the knowledge for a longer time. But what happens in the brain during the spacing effect, and why is taking breaks so beneficial for our memory? It is generally thought that during learning, neurons are activated and form new connections. In this way, the learned knowledge is stored and can be retrieved by reactivating the same set of neurons. However, we still know very little about how pauses positively influence this process—even though the spacing effect was described more than a century ago and occurs in almost all animals. Learning in a maze Annet Glas and Pieter Goltstein, neurobiologists in the team of Mark Hübener and Tobias Bonhoeffer, investigated this phenomenon in mice. To do this, the animals had to remember the position of a hidden chocolate piece in a maze. On three consecutive opportunities, they were allowed to explore the maze and find their reward—including pauses of varying lengths. "Mice that were trained with the longer intervals between learning phases were not able to remember the position of the chocolate as quickly," explains Annet Glas. "But on the next day, the longer the pauses, the better was the mice's memory." During the maze test, the researchers additionally measured the activity of neurons in the prefrontal cortex. This brain region is of particular interest for learning processes, as it is known for its role in complex thinking tasks. Accordingly, the scientists showed that inactivation of the prefrontal cortex impaired the mice's performance in the maze. "If three learning phases follow each other very quickly, we intuitively expected the same neurons to be activated," Pieter Goltstein says. "After all, it is the same experiment with the same information. However, after a long break, it would be conceivable that the brain interprets the following learning phase as a new event and processes it with different neurons." However, the researchers found exactly the opposite when they compared the neuronal activity during different learning phases. After short pauses, the activation pattern in the brain fluctuated more than compared to long pauses: In fast successive learning phases, the mice activated mostly different neurons. When taking longer breaks, the same neurons active during the first learning phase were used again later. Memory benefits from longer breaks Reactivating the same neurons could allow the brain to strengthen the connections between these cells in each learning phase—there is no need to start from scratch and establish the contacts first. "That's why we believe that memory benefits from longer breaks," says Pieter Goltstein. Thus, after more than a century, the study provides the first insights into the neuronal processes that explain the positive effect of learning breaks. With spaced learning, we may reach our goal more slowly, but we benefit from our knowledge for much longer. Hopefully, we won't have forgotten this by the time we take our next exam!     The flavonoid epicatechin inhibits progressive tau pathology in Alzheimer's University of Bath (UK), July 23, 2021 According to news reporting originating in Avon, United Kingdom, by NewsRx journalists, research stated, “Aggregation of the microtubule-associated protein tau into paired helical filaments (PHFs) and neurofibrillary tangles is a defining characteristic of Alzheimer's Disease. Various plant polyphenols disrupt tau aggregation in vitro but display poor bioavailability and low potency, challenging their therapeutic translation.”  Green tea, cocoa, blackberries and blueberries are high in epicatechin. The news reporters obtained a quote from the research from the University of Bath, “We previously reported that oral administration of the flavonoid (-)-epicatechin (EC) reduced Amyloid-beta (A beta) plaque pathology in APP/PS1 transgenic mice. Here, we investigated whether EC impacts on tau pathology, independent of actions on A beta, using rTg4510 mice expressing P301L mutant tau. 4 and 6.5 months old rTg4510 mice received EC (similar to 18 mg/day) or vehicle (ethanol) via drinking water for 21 days and the levels of total and phosphorylated tau were assessed. At 4 months, tau appeared as two bands of similar to 55 kDa, phosphorylated at Ser262 and Ser396 and was unaffected by exposure to EC. At 6.5 months an additional higher molecular weight form of tau was detected at similar to 64 kDa which was phosphorylated at Ser262, Ser396 and additionally at the AT8 sites, indicative of the presence of PHFs. EC consumption reduced the levels of the similar to 64 kDa tau species and inhibited phosphorylation at Ser262 and AT8 phosphoepitopes. Regulation of the key tau kinase glycogen synthase kinase 3 beta (GSK3 beta) by phosphorylation at Ser9 was not altered by exposure to EC in mice or primary neurons. Furthermore, EC did not significantly inhibit GSK3 beta activity at physiologically-relevant concentrations in a cell free assay.” According to the news reporters, the research concluded: “Therefore, a 21-day intervention with EC inhibits or reverses the development of tau pathology in rTg4510 mice independently of direct inhibition of GSK3 beta.”     Fruit compound may have potential to prevent and treat Parkinson's disease   Johns Hopkins University, July 29, 2021 Johns Hopkins Medicine researchers say they have added to evidence that the compound farnesol, found naturally in herbs, and berries and other fruits, prevents and reverses brain damage linked to Parkinson's disease in mouse studies. he compound, used in flavorings and perfume-making, can prevent the loss of neurons that produce dopamine in the brains of mice by deactivating PARIS, a key proteininvolved in the disease's progression. Loss of such neurons affects movement and cognition, leading to hallmark symptoms of Parkinson's disease such as tremors, muscle rigidity, confusion and dementia. Farnesol's ability to block PARIS, say the researchers, could guide development of new Parkinson's disease interventions that specifically target this protein. "Our experiments showed that farnesol both significantly prevented the loss of dopamine neurons and reversed behavioral deficits in mice, indicating its promise as a potential drug treatment to prevent Parkinson's disease," says Ted Dawson, M.D., Ph.D., director of the Johns Hopkins Institute for Cell Engineering and professor of neurology at the Johns Hopkins University School of Medicine. Results of the new study, published July 28, in Science Translational Medicine, detail how the researchers identified farnesol's potential by screening a large library of drugs to find those that inhibited PARIS. In the brains of people with Parkinson's disease, a buildup of PARIS slows down the manufacture of the protective protein PGC-1alpha. The protein shields brain cells from damaging reactive oxygen molecules that accumulate in the brain. Without PGC-1alpha, dopamine neurons die off, leading to the cognitive and physical changes associated with Parkinson's disease. To study whether farnesol could protect brains from the effects of PARIS accumulation, the researchers fed mice either a farnesol-supplemented diet or a regular mouse diet for one week. Then, the researchers administered pre-formed fibrils of the protein alpha-synuclein, which is associated with the effects of Parkinson's disease in the brain. The researchers found that the mice fed the farnesol diet performed better on a strength and coordination test designed to detect advancement of Parkinson's disease symptoms. On average, the mice performed 100% better than mice injected with alpha-synuclein, but fed a regular diet. When the researchers later studied brain tissue of mice in the two groups, they found that the mice fed a farnesol-supplemented diet had twice as many healthy dopamine neurons than mice not fed the farnesol-enriched diet. The farnesol-fed mice also had approximately 55% more of the protective protein PGC-1alpha in their brains than the untreated mice. In chemical experiments, the researchers confirmed that farnesol binds to PARIS, changing the protein's shape so that it can no longer interfere with PGC-1alpha production. While farnesol is naturally produced, synthetic versions are used in commerce, and the amounts people get through diet is unclear. The researchers caution that safe doses of farnesol for humans have not yet been determined, and that only carefully controlled clinical trials can do so. Though more research is needed, Dawson and his team hope farnesol can someday be used to create treatments that prevent or reverse brain damage caused by Parkinson's disease.   Plant compounds reveal anticancer mechanisms Russian Academy of Sciences, July 28 2021.  Research published on June 9, 2021 in Scientific Reportsexplored mechanisms involved in the cancer protective effects of 30 compounds derived from fruits and vegetables. The researchers hope that their findings will contribute to the formulation of new drugs that will have fewer side effects than drugs currently in use.   “To create potent new drugs that will target only the tumor, it was necessary to determine how dietary compounds affect cell proteins in the prevention and treatment of cancer,” explained coauthor Grigory Zyryanov, who is a professor at the Russian Academy of Sciences. “Therefore, by modeling molecular mechanisms, we figured out how substances bind to proteins. This allowed us to determine the pool of therapeutic targets that the drugs will subsequently target. For example, these are anti-apoptotic (prevent apoptosis) and pro-apoptotic (induce apoptosis) proteins, protein kinases, and others. But a key drug target is phosphatidylinositol-3-kinase . . . This enzyme influences mutations in cancer, rearrangement, and amplification of genes.” Compounds investigated in the study included emodin, eugenol, gingerol, sulforaphane, linalool, catechin, oleanolic acid, ursolic acid, curcumin, yakuchinone-A, pinusolide, alpha-boswellic acid, oleandrin, sesquiterpene lactone-326, resveratrol, triterpenoid, beta-boswellic acid, anethole, capsaicin, glycolic acid, quercetin, genistein, ellagic acid, flavopiridol, zerumbone, garcinol, guggulsterone, parthenolide, halogenated monoterpenes and silibinin. Of these compounds, silibinin, flavopiridol, oleandrin, ursolic acid, alpha-boswellic acid, beta-boswellic acid, triterpenoid, guggulsterone and oleanolic acid had the greatest binding affinity with phosphatidylinositol-3-kinase alpha (P13K), which is involved in functions that can contribute to cancer. Other targets identified as binding with various compounds included PKC-η, Ras and H-Ras.  “We assumed that the foods we selected for the study had anti-cancer properties, but this needed to be verified,” Dr Zyryanov noted. “As a result, we found out that diseased cells stop development under the influence of certain combinations of food compounds.”     Meta-analysis supports potential of omega-3s for ADHD Kings College London, July 31, 2021 Omega-3s fatty acid supplements may improve symptoms and cognitive performance in children and adolescents with attention deficit hyperactivity disorder (ADHD), according to a meta-analysis of gold standard clinical trials. Data from seven clinical trials involving over 500 children and adolescents indicated that omega-3s were associated with improvements in clinical symptoms of ADHD, while data from three clinical trials involving over 200 children and adolescents indicated a positive impact on cognitive measures associated with attention. “[W]e provide strong evidence supporting a role for n3-PUFAs deficiency in ADHD, and for advocating n-3 PUFAs supplementation as a clinically relevant intervention in this group, especially if guided by a biomarker-based personalization approach,” wrote the authors, led by Jane Pei-Chen Chang from King's College London, in Neuropsychopharmacology . Boosting EPA/DHA intakes Commenting independently on the meta-analysis, Harry Rice, PhD, VP of regulatory & scientific affairs for the Global Organization for EPA and DHA Omega-3s (GOED): “In the past, I've been lukewarm on whether or not increasing EPA/DHA intake benefits children with ADHD. Results from this meta-analysis put me a little closer to believing. “Minimally, given the low side effect profile of omega-3s versus the drugs of choice to treat ADHD, I would highly recommend first increasing intake of EPA/DHA. This is particularly true if a child doesn't eat at least two servings of fatty fish a week or doesn't take an omega-3 supplement on a regular basis.” Meta-analysis details The new meta-analysis was performed using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines and used established scientific literature databases to identify appropriate studies for inclusion. Data from seven randomized controlled trials (RCTs) with 534 young people indicated that that omega-s3 supplementation significantly improved inattention and hyperactivity symptoms, according to parental reports. Additional analysis revealed that the improvements in hyperactivity were only observed when doses of EPA (eicosapentaenoic acid) of 500 mg/day or more were used. Interesting, the researchers did not find improvements in hyperactivity and inattention when they looked at teacher's reports, unlike what was reported by parents. Omega-3 supplements were associated with improvements in select measures of cognitive performance, said the researchers. “N-3 PUFAs are crucial for optimal neurotransmitter function: for example, incorporating more EPA and DHA in the cell membrane can increase cholesterol efflux, modulate lipid raft clustering and disruption, and affect the function of the dopamine transporter (DAT), which in turn may affect attention and executive function by regulating synaptic dopamine levels,” wrote the researchers. Omega-3 levels Data from case-control studies were also collected to assess if omega-3 levels were also associated with ADHD, with results indicating that children and adolescents with ADHD had lower levels of EPA, DHA (docosahexaenoic acid),and total omega-3s. “In the context of ‘personalised medicine', it is tempting to speculate that a subpopulation of youth with ADHD and with low levels of n-3 PUFAs may respond better to n-3 PUFAs supplementation, but there are no studies to date attempting this stratification approach,” wrote the researchers. “However, we have [previously] shown that individuals at genetic risk of developing depression in the context of the immune challenge, interferon-alpha (IFN-alpha), have lower levels of RBCs n3-PUFAs, and that n-3 PUFAs supplementation prevents the onset of IFN-alpha-induced depression, arguably by replenishing the endogenously low anti-inflammatory PUFAs in the ‘at risk' individuals.”

From our physical appearance to our body's mechanical functions, our whole being is encoded into our genes and kept in each cell that makes us. These basic biological units have their system to keep everything functioning and our body alive and moving. They have housekeeping functions: cells perform autophagy to get rid of accumulated waste materials. Maximising the effects of these processes can help in increasing your longevity. When the autophagic processes fail, it can damage important parts of the cell such as the DNA and accelerate ageing. In this episode, Dr Elena Seranova explains the science behind autophagy and how it connects to NAD and sirtuin genes. She also shares her own experience and research on using this knowledge to live a longer, fuller life. Join us in this episode to learn more about autophagy and how this process is useful in increasing your longevity and giving you a boost in life.   Get Customised Guidance for Your Genetic Make-Up For our epigenetics health program that is all about optimising your fitness, lifestyle, nutrition and mind performance to your particular genes, go to https://www.lisatamati.com/page/epigenetics-and-health-coaching/. You can also join our free live webinar on epigenetics.   Online Coaching for Runners Go to www.runninghotcoaching.com for our online run training coaching.   Consult with Me If you would like to work with me one to one on anything from your mindset, to head injuries, to biohacking your health, to optimal performance or executive coaching, please book a consultation here: https://shop.lisatamati.com/collections/consultations.    Order My Books My latest book Relentless chronicles the inspiring journey about how my mother and I defied the odds after an aneurysm left my mum Isobel with massive brain damage at age 74. The medical professionals told me there was absolutely no hope of any quality of life again, but I used every mindset tool, years of research, and incredible tenacity to prove them wrong and bring my mother back to full health within 3 years. Get your copy here: http://relentlessbook.lisatamati.com/  For my other two best-selling books Running Hot and Running to Extremes, chronicling my ultrarunning adventures and expeditions all around the world, go to https://shop.lisatamati.com/collections/books.    Here are three reasons why you should listen to the full episode: Discover how cells stay healthy. Find out Dr Elena's tips and advice in increasing your longevity by activating autophagy. Learn more about the science behind NMN supplements and their revitalising effect.   Resources Read about Dave Asprey’s work around the keto diet. Lifespan by Dr David Sinclair Pushing the Limits Ep 170: The Search for the Perfect Protein with Dr David Minkoff Visit NMNBio if you want to learn more about NMN supplements in New Zealand.   Episode Highlights [04:33] Getting to Know Dr Elena Seranova Dr Elena Seranova is an interdisciplinary scientist.  She holds a degree in Psychology, MSc Translational Neuroscience, and a PhD focusing on autophagy and cell biology. With her expertise in her field, she co-founded a biotech startup and is now the the founder of NMN Bio in the United Kingdom.  [06:06] What Is Autophagy? Autophagy is a catabolic pathway that degrades unwanted materials within the cell. The cell needs to avoid the build-up of unnecessary materials. There are different pathways for activation like mTOR (mammalian target of rapamycin) and PI3K (Phosphoinositide 3-kinase). Impairments at various stages of autophagy lead to its failure and cell death. Once autophagy fails, apoptosis, or programmed cell death, can activate. [11:25] NAD as Fuel for Sirtuin and PARP PARP and sirtuin are different classes of enzymes that use NAD for multiple vital processes, including DNA repair (both) and gene expression (sirtuin). Sirtuin 1 is one of the pathways that can initiate autophagy. initiates the autophagic process. When DNA is damaged, PARP activates and depletes NAD stores. The decrease in NAD levels inhibits sirtuin's ability to carry out its functions, including autophagy, accelerating a cell's death. [15:34] What Is NAD? Nicotinamide adenine dinucleotide or NAD is a substrate for enzymes. It plays a vital role in different reactions within the cell. You can supplement NAD levels using boosters such as nicotinamide mononucleotide (NMN). [18:44] mTOR-Independent Pathways Activation of autophagy using Sirtuin 1 is an mTOR-independent pathway. It is essential not to activate autophagy through mTOR pathways. mTOR is responsible for cell growth and translation. [25:04] How to Activate mTOR-Independent Autophagy Autophagy is dependent on nutrient starvation. Intermittent fasting can activate it. A generally healthy lifestyle includes supplement intake, proper sleep, and healthy foods. Avoid too much sunlight because it damages the skin and DNA. It activates PARP enzymes. Take a tablespoon of extra virgin olive oil which contains oleic acid. This good fat has the same effects as resveratrol. Induce artificial stress through cryotherapy, saunas, and exercise to activate sirtuin 1.  [33:22] Why Did Dr Elena Decide to Bring NMN to the Market? She came across NAD and NMN during her doctorate studies and saw their importance. Her own experiences showed better focus and energy levels after taking NMN supplements. There are not enough reliable suppliers that offer good regulation and quality control. She wanted to provide the best for herself and her family. NMN Bio has complete transparency on suppliers and quality regulation [38:28] Can Weight Loss Occur from Taking NMN? Lisa Tamati's experience in taking NMN shows fat loss but no muscle loss. Dr Elena's mice studies show evidence that NMN can improve insulin sensitivity and lipid metabolism. [40:42] Do NMN Supplements Have Any Downsides? They found no side effects in studies with mice despite an increase in dosage. Current studies are still ongoing to determine the ideal dosage for humans. Most people take 500mg to 1g. Doubling this still shows good tolerance. Take supplements in the morning as NAD affects the circadian rhythm. [53:57] How Do Autophagy, NAD and Sirtuin Genes Fit Together? Autophagy recycles various damaged organelles. Sirtuin 1 genes activate autophagy and mitophagy. NAD functions as a substrate for sirtuin enzymes to work. NMN supplements can increase NAD levels. Listen to the episode for the full explanation of how these three work together. [58:43] Can NMN be Taken as an Infusion? This is not something that Dr Elena has studied in-depth and she is curious as well about how viable this procedure will be.  There is a low concentration of energy in intravenous injections, but it's present. Oral administration is more reliable in giving boosts and it costs cheaper. [01:00:01] Do Antioxidants Help in Increasing Your Longevity? Studies have shown that antioxidants don't suppress ageing. Lifestyle intervention and autophagy activation are proven ways to slow ageing.   7 Powerful Quotes  ‘Lysosome is another acidic organelle that contains acid hydrolases that are able to digest this cargo...and if it doesn't work, well, the cell is basically in trouble because you have all this garbage floating around, and there is nothing to remove them. So this is why autophagy is important. ‘When things are preserved across species, then that gives a scientist an indication that this is probably a very important biological function’. ‘I think that in order for your body to function properly, you really need to have a kind of a healthy routine in general’. ‘So I think that when it comes to being healthy, and activating your autophagy levels, and having a healthy lifestyle in general, you need to start with the basics first.’ ‘Another small tip that I can give is to actually avoid sunlight, which is something that people don't really consider. What happens when we're exposed to sunlight, when our skin is exposed to sunlight for prolonged periods of time, we start getting the DNA damage’. “The important part is not to just increase your age, it’s to increase your healthspan.”  ‘And if you have if your mitochondria are not healthy, and they're dying, and you're not having enough mitochondria in your cells, then you are going to be sick.’   About Dr Elena Seranova Dr Elena Seranova is a scientist, serial entrepreneur and business mentor who has founded multiple innovative biotechnological businesses. She first studied at the University of Ioannina with a major in Psychology. She started a private practice before developing an interest in neuroscience. She continued her studies and earned her Master’s Degree in Translational Neuroscience at the University of Sheffield. She now also holds a Doctorate Degree in Stem Cell Biology and Autophagy from the University of Birmingham. Her expertise in these fields has led her to become the co-founder of a biotech start-up, SkyLab Bio. She has written a number of peer-reviewed articles and multiple research articles on autophagy throughout her career. Aside from these accomplishments, she started her own business, NMN Bio. Her own experiences with the use of supplements have inspired her to expand the market to supply the public with cutting-edge anti-ageing supplements. NMN Bio reaches New Zealand, UK, and Europe.  Dr Elena found her passion for drug discovery and autophagy. She has endeavoured to share this with the public through her research and work as an entrepreneur.  To learn more about Dr Elena and her work, visit her website.    Enjoyed This Podcast? If you did, be sure to subscribe and share it with your friends! Post a review and share it! If you enjoyed tuning in, then leave us a review. You can also share this with your family and friends, especially those interested in increasing their longevity, so that they can practice the activation of autophagy in their lifestyle. Have any questions? You can contact me through email (support@lisatamati.com) or find me on Facebook, Twitter, Instagram and YouTube. For more episode updates, visit my website. You may also tune in on Apple Podcasts. To pushing the limits, Lisa   Transcript Of The Podcast Welcome to Pushing the Limits, the show that helps you reach your full potential, with your host Lisa Tamati, brought to you by lisatamati.com. Lisa Tamati: Well, hi everyone and welcome back to Pushing the Limits. This week I have Dr. Elena Seranova, who has already been a guest on the show. And today, we're talking about autophagy and NAD, and the interplay between these two. Now that might sound extremely boring, but it isn't. It's all about longevity and anti-aging. So, we're going to be talking about the science between about NAD precursors and the sirtuin genes, and how to upregulate the sirtuin genes, and all about autophagy, which is really the recycling of old and damaged parts and proteins of a cell that need to be gotten rid of. So, it's a bit like having a good garbage disposal unit happening. And there are many ways to activate autophagy, which we go into in this episode. We talk about intermittent fasting, we talk about cold and hot and hormetic stressors like exercise and yes, of course fasting. But also, mTOR independent pathways to activate autophagy, it'll all be revealed in this interview.  Now this does get a little bit technical in the first 20 minutes or so. But hang in there and listen to this a couple of times. Because if you want to slow down aging, if you want to slow down the generative decline of your body and you want to have a long and healthy lifestyle, then this stuff is really, really worth paying attention to and trying to understand.  We talk about NMN, which is nicotinamide mononucleotide, which is a supplement that is now available, is a longevity compound to upregulate the sirtuin genes. And we're really lucky to check that out. You can go and find that supplement, which has been made and produced by Dr. Elena Seranova and her company, NMN Bio. So if you head hop over to nmnbio.nz, we're now importing this into the country. So, this is one way that you can really fight aging and degenerative decline that we all fear and don't want.  And when you listen to this episode and really listen to it a couple of times, you'll understand some of the incredible anti-aging things that are coming down. This is not pseudoscience. Dr. Elena is one of the most amazing neuroscientists out there. So please listen to this episode, enjoy it, get a lot out of it and get the takeaway. So, if you don't understand some of the terminology, don't worry, keep going. And by the end, you'll start to pick up certain bits and pieces. And if you listen to it again, you'll be able to pick up a little bit more and a little bit more. And at the end of the day, it's about the takeaways, what can you do to slow aging down and all that information is in there. So, I hope you enjoy this episode with Dr. Elena.  Before we go over to that we are all about health optimisation, high performance, athletic performance. So, if you need any help with any of those areas of your life, please reach out to us, Support at lisatamati.com. Go and check out our website, lisatamati.com. You'll find all our programs, our Epigenetics Programs, our online run training system that's customised and personalised totally to you, and check out what we do. We love helping you be the best version of yourself that you can be. Now over to the show with Dr. Elena Servanova.  Lisa: Well, hi everyone. Lisa Tamati here and very excited to have you hopefully join us this morning. It's 7:30am in the morning here in New Zealand. And where Dr. Elena Seranova is, it's very late at night. How are you doing, Dr. Elena?   Dr Elena Seranova: Good, good. How are you? Happy to be here again. Lisa: Yeah, very excited for today's topic. So, we're going to be doing a discussion around autophagy and NAD boosters and sirtuin genes. So it’s going to be a really interesting discussion that is really beneficial for you if you want to know how to live longer, live healthier, and optimise your body and your mind and your potential. So, Dr. Elena, can you just tell us briefly a little bit about yourself?  Dr Elena: Sure. So, I started my journey as a psychologist. So I'm an interdisciplinary scientist. I majored in psychology at first and then I had my own private practise for five years which turned out to be a successful wellness centre. And I really got fascinated by neuroscience and the brain. And for this reason, at first I started studying the brain myself and then I found an amazing master's degree at the University of Sheffield in Translational Neuroscience, which basically combined the research and neurodegeneration with applications that could translate into therapeutics. So, this is what translational neuroscience means, is basically the combination and the outcome of the research—the hardcore biology research that can be utilised for therapeutic approaches and patients.  I really enjoyed that. So that was quite cool, being in the lab and doing molecular biology experiments and so on. So, I kind of fell in love with the lab, and I decided to do a PhD as well. I continued my studies in autophagy and stem cell biology and it was quite challenging, but at the same time, I really enjoyed it. And I can definitely say that science is a big part of my life. Lisa: Definitely your thing. Okay, so autophagy and stem cells. So in relation to neurodegenerative diseases in that case? Okay, but what is autophagy? Because a lot of people will be listening to it and go, ‘What the heck is that big word, autophagy’? It's sort of big word in biohacking circles, but perhaps not in the general public. Can you explain what autophagy is exactly?  Dr Elena: Yes, sure. So, autophagy is a catabolic pathway that degrades dysfunctional organelles in the cell or protein pro aggregates. So, any material that is basically unwanted in the cell, autophagy can degrade. It's like the stomach of the cell.  Lisa: So, it’s like eating it? It's eating, sort of...  Dr Elena: Yeah, exactly. And what happens when autophagy is activated, we actually have the formation of the so-called phagophore, which is a membrane structure that basically engulfs different organelles and materials that need to be degraded to form the so called autophagosome, which is a round organelle that basically has this cargo that needs to be digested. That eventually fuses with lysosome.  And lysosome is another acidic organelle that contains acidic hydrolases that are able to digest this cargo. And this process is very essential for the cell, it’s very vital. It's evolutionary conserved in all species, from yeast to models. And if it doesn't work well, the cell is basically in trouble because you have all this garbage... Lisa: Floating around.  Dr Elena: ...floating around and there is nothing to remove them. So, this is why autophagy is important. And we have different pathways that autophagy can be activated through as well. So, one of those pathways is mTOR, mechanistic target of rapamycin. And then we have other pathways that can activate this process such as AMPK, GSK3, and so on.  Lisa: So is this like, sorry to interrupt, but like because I know that people out there might be like, ‘Wow, that's a lot of big words and a lot of information’. So, is it like that the cell has to do a housecleaning, and it's got stuff inside the cell that is not working optimally, and needs to be gotten rid of, or is it the whole cell? So, it's not apoptosis. So it's not where the host is disintegrating? Dr Elena: No. Yeah, it's actually a—it's a pre-apoptotic pathway. So, before apoptosis is activated, we have autophagy. And if autophagy fails in what it needs to do, then we have activation of some apoptotic pathways. So, it's one step before that. And if everything goes well, and autophagy is functional—and by the way, in different diseases, we might be having different autophagy impairments at different stages of autophagy. So, it's either the initial phagophore formation, for instance, that it's not working well, and it can’t engulf the cargo, or it's insulator stages of autophagy, such as the acidic hydrolysis and the lysosomes that are actually not that acidic. So their pH is not acidic enough to digest the cargo. So, we might be having different defects in the autophagy pathway in different diseases. Lisa: That leads to apoptosis. Am I right?  Dr Elena: And yeah, if autophagy is not doing its work correctly, then eventually we will have apoptosis. And actually, this is what we're seeing in in vitro models of neurodegenerative diseases as well. So, for instance, if autophagy is not working well.  And we have, let's say, dysfunctional organelles, such as mitochondria—dysfunctional mitochondria that are not working well. Let's say they are depolarised. And there is an excess production of reactive oxygen species going on. Now, if nothing can degrade these dysfunctional mitochondria, you'll keep on having this accumulation of reactive oxygen species, which eventually will lead to DNA damage and deactivation of PARPs. And it's basically a death spiral that will keep on leading the cell towards death.  Lisa: Okay, so what is a PARP? You mentioned PARP there. And just for the listeners, too. So, apoptosis is basically cell death, programmed cell death. So, this is not—what's the other one necrotic or something?  Dr Elena: Necrosis?  Lisa: Yeah, necrosis, where the cell dies for—necrosis. But this is sort of a natural programmed cell death. But we only want that if we're actually renewing the cells and we are wanting new stuff. So, before that, the body tries to do this autophagy process, is that how it works? And then what so what is PARP? What is PARP, because that’s the word again... Dr Elena: PARPs are a class of enzymes, and in order for them to function, they need a molecule called NAD, so nicotinamide adenine dinucleotide. And they're actually competing for NAD in the cell. And whenever we have increased DNA damage, we would have the PARP activation as well. And this would lead to NAD depletion, which kind of brings me to my next point about what other enzymes consume NAD. And one of those enzymes are sirtuins, which are the so-called longevity genes that are basically responsible for multiple processes in the cell, including epigenetic regulation of gene expression.  So, they do—because sirtuins are a class of enzymes that are also dependent on NAD, and they're all the deacetylase enzymes, meaning that they remove acetyl groups from the DNA. And as a result, they control which genes will be expressed in which tissues, which is very crucial for the cellular identity and for the proper function of different cells.  So, sirtuins in a healthy cell, so sirtuin should be upregulated and they should be having this housekeeping gene—housekeeping function where they basically control what's going on with the DNA repair and also with the gene expression as well. And if we do have—when we do start having impaired autophagy, and let's say there is increased reactive oxygen species, because there are increased dysfunctional mitochondria in the cell, you will have in more activation of PARPs, and all of the NAD will start being drained from... And sirtuin will not have enough energy to function. So, those are actually quite an elegant interplay between autophagy and NAD and sirtuins.  Lisa: Okay. Okay, can I just want to like put that back to you, so that we can slow down because we are going technical quite fast. And I think a lot of people might be like, ‘What the heck are they talking about’? So, the sirtuin genes, basically longevity genes, and then one of the jobs is DNA repair. And another of the jobs is to say which genes are actually being activated right now. And these sirtuin genes are also responsible, I think, for cell replication, is that correct? Dr Elena: The sirtuins are responsible for multiple functions, directly or indirectly. So, for instance, the sirtuin 3 gene is also responsible for mitochondrial biogenesis. And it's implicated in the amount of mitochondria that are being produced by the cell, which is related to cell replication eventually, because you do need to have enough ATP levels to replicate. Lisa: Right. Yeah. So, this has definitely to do with ATP production as well and mitochondrial health. So, these are doing all of these jobs, the sirtuin genes, they're very, very crucial genes in our genome. And these are preserved across every species, I believe? Every species on the planet?  Dr Elena: Yeah. Also from yeast to humans, it's also—sirtuin genes are preserved very well. Lisa: And when things are preserved across species, then that gives a scientist an indication that this is probably a very important biological function and we need to have a look at this one because it's—from what I understand.  Okay, so when you have activated PARP because you're not doing autophagy well and there's things going wrong, it's taking the NAD. So NAD is basically like a fuel source that both the sirtuin genes. And when PARP is activated, it's using to fuel its job. And so, this is competition for competing fuel sources. So, like if you imagine, you've only got one tank of fuel for your car, but you've got to go in two different directions and do two different jobs. You go, ‘How am I going to divide up my energy’? So, then it becomes important as to how much NAD we have in the body? So, what is NAD again? That says nicotinamide adenine dinucleotide? But what is that and how does it work?  Dr Elena: Yeah, it does serve as a substrate for all of these enzymes, including sirtuins and PARPS and is basically a master regulator of metabolism. So, it's a very important molecule and it serves as—without NAD, the cell is not able to function properly just because this crucial molecule is implicated in so many different reactions. So, NAD is found in all living cells and organisms. This is also evolutionary conserved across species. And it exists in two forms, NADH and NAD+, which is the reduced and the oxidised form, respectively. And both of them are important. And both of them are implicated in multiple cellular reactions.  Lisa: Is it going backwards and forwards in a cycle, NADH, NAD+, by donating electrons back and forth, sort of thing?  Dr Elena: Yeah. Through electron transport chain in the mitochondria, yeah. So, this is why it's so important. And so, what we're seeing now in the latest advancements in longevity research is that we actually can supplement with different precursors of NAD, such as nicotinamide mononucleotide, for instance, NMN. And this is the supplement that my company...  Lisa: See, you've now got that available on the market because this is such a crucial thing. Dr Elena: Exactly. And I think that it's really interesting to also say that when it comes to the interaction between autophagy and sirtuins, there is also another regulation of autophagy there. So sirtuin 1 is actually responsible for activating some transcription factors such as TFEB and FoxO3 that have to do with initiation of the autophagy process.  So, for this reason, when we do have dropping levels of NAD, decreasing levels of NAD, and there is not enough NAD for sirtuins to do their job. And let's say again—let's talk about that previous example in neurodegeneration when you have increased reactive oxygen species, and you have increased the level of stress and oxidative stress and decreased activity of sirtuins.  And not only the situation is already bad, but because sirtuin 1 doesn't have enough energy to function and to activate the TFEB and the FoxO3 transcription factors to initiate autophagy, now you have all of this dysfunctional mitochondria floating around and autophagy starts being impaired as well because we activated enough. So, it's a negative feedback loop which actually accelerates the scenario where the cell is going towards cell death, basically. Lisa: So that means like, if you don't have enough NAD, then your sirtuin 1 gene is not going to be able to initiate autophagy and clean up the cell and you're going to have dysfunctional mitochondria. Is that independent of the mTOR pathway? Or is that—am I getting confused?  Dr Elena: So, okay. So, good question. So, what happens is there are some molecules that activates sirtuins. So, for instance, sirtuin 1 is activated by resveratrol, and this is something that has been demonstrated many years ago. So, when you have sirtuin 1 dependent activation of autophagy, you will be having it through an mTOR independent pathway. Lisa: So it's a fasting mimetic resveratrol.  Dr Elena: Yeah, yeah. Yeah, absolutely. So, because we now know that the mTOR activity is not affected by intake of resveratrol. And this is quite crucial because actually, even if we want to activate autophagy, we shouldn’t do it through the mTOR pathway, this is not the preferred way, because mTOR is also responsible for growth and translation in the cell. So, this is not—it's also quite a key player in the cell. So it's a serine threonine kinase, and you actually don't want it to be activated at all times because this may lead other conditions. So, what we're focusing on at the moment is to find molecules that can activate autophagy in an mTOR independent manner. Lisa: Okay, so. So if the mTOR—cause MTOR is usually what's for growth it’s anabolic, it's causing growth. So for example, a bodybuilder goes to the gym, they're in an anabolic state, they are in an mTOR growth state. And when you have autophagy, that's sort of the opposite. So, it's a catabolic state where it's starting to eat itself. So, it’s mTOR, most people like do fasting for that reason to activate autophagy? Dr Elena: Yeah, this is another good point there. So, when we're fasting, and there is actually conflicting evidence out there as to when autophagy is fully activated. Usually, people say that around 24 hours, you start having the autophagy activation. There are others that swear by the ketogenic diet, and say that if you don't consume any carbs, you will get autophagy activation anyway. However, from what other researchers have found is that, if you are in a ketogenic diet, and you do consume meat, it depends on what kind of meat you consume that will either activate autophagy or not. And it all has to do with levels of different amino acids in the cell because autophagy is quite sensitive to nutrients and to nutrient starvation to be activated. If you have an abundance of amino acids, again, it will not be activated.  So, for instance, one amino acid that activates autophagy very well is leucine. And if you're eating certain meat that are rich in leucine, this is probably not good for your autophagic state. Something else to keep in mind, and I've heard, I think it was Dave Asprey saying that if you can manage to be on under 15 grams of protein per day, you will probably keep the autophagy going.  Lisa: Because a lot of people on keto think I can eat a lot of protein, which is a mistake, really. It isn't about having—that's interesting, because I had Dr. David Minkoff on my podcast, Pushing the Limits a while ago, and he has a product called PerfectAmino, which is really a 99% usable form of amino acids and combination. And I was interested, ‘Well hang on, if I'm heading there, which is going a lot of good things in the body. But is that going to inhibit my mTOR, or autophagy’? Sorry, because I've got too much leucine in there? Dr Elena: This is a very good point for all of this process food as well. So, for instance, there are some ready meals you can get or some protein bars that claim to have all the low carb and everything. And then they slam a badge on their pack saying that it's vegan as well. But then, why is it vegan if it has all the amino acids because that's one of the selling points when you're actually on a vegan diet, or you have some days where you are on a vegan diet. You want to get yourself in a state of partial amino acid depletion to get this beneficial effect of enhanced autophagy. And on intercellular toxins and so on.  Lisa: Right, so for certain periods of time, you want to do this, and it's a cycling thing, you don't want to be completely deficient of aminos for too long because then your body will start to break down. Dr Elena: This is what I do personally as well. So, during the week, so I am a fan of cattle/carnivore diet. So, this diet is quite comfortable for me and I enjoyed it quite a lot. But then during my week I try to have some days where I'm either vegetarian or vegan, just because I want to have those benefits.  Lisa: Yeah. Up and down. And then this seems to be a theme in biology all the time is that it's not one thing. It's not staying on keto for ever and ever, amen. It's about doing cyclic keto or cyclic vegan and it’s cyclic. And our body loves this push and pull—when there’s recovery and there’s growth and then clean up phase, growth clean up. So autophagy can be activated through fasting. It can also be active through having resveratrol and upregulating the sirtuin 1 gene, how else can we activate autophagy? Dr Elena: So there are different ways, there are different things you can really implement in order to activate autophagy. And I think that it all has to do with how you build your lifestyle in general. So, I think that in order for your body to function properly, you really need to have a kind of a healthy routine in general. And an analogy that I can give you there is that there are people that would buy a couple of supplements, and then they would be so proud of it. And then they would say, ‘Oh, yeah, but I'm taking those supplements now, and I'm so healthy’. And then their biorhythms are all off. They sleep at 5am every day. And they're eating crappy foods or super processed foods.  Lisa: Yeah, it’s not going to work.  Dr Elena: It’s all good. So, I think that when it comes to being healthy and activating your autophagy levels and having a healthy lifestyle in general, you need to start with the basics first.  So, the intermittent fasting is definitely the first step to take in order to become a bit healthier. And from the research that I'm reading, and from the things that I'm implementing, I definitely believe that both anecdotal and scientific evidence point towards the fact that intermittent fasting is actually the way to go. I mean, there are conflicting opinions out there and there are pros and cons in every diet, and so on. And I get that. But I personally believe that with intermittent fasting, if you try to narrow down the window where you're uptaking food, this is very, very good for you. So, this is step number one.  But then again, so either you're trying to raise your NAD levels, or you're trying to activate your autophagy, because those pathways are quite intertwined. And what you eventually want to do is you want to have increased levels of sirtuin, and sirtuin 1 in particular, and sirtuin 3, of course, and so on. And for this reason, in order to preserve this pool of NAD that is available for the sirtuin 1 to activate itself and activate the autophagy pathway.  Another small tip that I can give is to actually avoid sunlight, which is something that people don't really consider. But what happens when we're exposed to sunlight, when our skin is exposed to sunlight for prolonged periods of time, we start getting the DNA damage. And when you get the DNA damage, you have PARP activation, and then again, you NAD pool... Lisa: Wow. I never connected those dots. That's really interesting. So, because—I mean, we need sun. We need sun for vitamin D and for our mood and all that sort of stuff. So, you're not saying don't have any sun.  Dr Elena: Yeah, sure.  Lisa: But because the sun is causing DNA damage, it's going to cause more PARP activation, it’s going to have the sirtuin genes going to repair the DNA, that's going to use up the body's resources is what you're saying.  Okay, wow, that makes sense. Makes sense. And then by the same token, like things like smoking that breaks DNA, like no tomorrow. This is why smoking ages you is because of all the DNA breaks. And this is why, when you're in the sun for hours every day, you get wrinkly skin and you get collagen lost and all the rest of the things that are happening. So, anything that's going to be causing DNA breaks is going to cause you to age quicker.  Dr Elena: Exactly.  Lisa: Using up the resources basically. Wow, okay. Dr Elena: So it's obviously—you don't have to become a vampire and dissipate walk in the sun when you want to go somewhere. But sunbathing for hours is definitely not something you want to do with—to get your body go through, basically. So that's another tip.  And then something else, really, really simple that can be implemented on a daily basis in order to maintain your sirtuin levels, and as a result, your autophagy levels, and your NAD levels is also to take a tablespoon of extra virgin olive oil, which contains oleic acid. And it basically does the same job as resveratrol. And it's interesting—I think that there's been a recent research article out that shows that like oleic acid might even be more efficient than resveratrol, in terms of activating sirtuin 1, which I think it's really, really cool.  Lisa: So yeah. Well, combine the two. I do. Dr Elena: Yeah, absolutely. You can do that. And then, you need to make sure that the extra virgin olive oil is actually of a very good quality because there is a bunch of...  Lisa: There is a bunch of rubbish out there. So, make sure it's from an orchard that you know, it's cold pressed, it's all those extra virgin, it's all that sort of good stuff. And not—how do they do it with solvents and stuff? Or that it's come from multiple orchards and being cut with other oils. It's a really, really important point. And then oleic acid does so much good things in the body. But isn’t that fat, Elena? Like lots of people are like, in their minds are going, ‘But oil is fat. It’s the same with MCT oil. Isn’t that going to make you fat when you eat fat’? Just going to put that around.  Dr Elena: There are good fats and there are bad fats. So, olive oil is good fat. MCT oil is a good fat. Avocado is a good fat. So, not all thoughts are made equal. So, this is definitely something important to keep in mind, especially with a good quality extra virgin olive oil.  Lisa: Because each one of our cells is a membrane that has a phospholipid, isn't it? So we need that,  actually, this building of ourselves into the integrity. Dr Elena: We have a phospholipid layer in the brain as well. And this is why we actually supplement with omega 3 fatty acids, because this is what it does. So, this is what omega 3 fatty acids do. They go into the phospholipid membrane, and then they basically... Lisa: Make the integrity of that membrane better. Dr Elena: Yeah, they contribute to the healthy phospholipid layer in the brain.  Lisa: So that's why it's very important for neurodegeneration to have omega 3s going in and again, people get quality omega 3s. Not your cheap supermarket ones that are perhaps oxidised and have been sitting on the shelves for six months. So really important to get a reputable source here. And omega 3 is of course in fishes as well, and krill, and so on.  Okay, so but is there a downside to fat? Because I studied epigenetics and a lot of people's profiles come back with don't have too many fats. And it's been one of those things in my head is like ‘Why would some people not come back with you shouldn't have too much fat’? I mean, there are things like gall bladders been removed. That's a pretty specific thing. But is there a genetic component? And probably not your wheelhouse, really, but is there a genetic component to your ability to process fat? Dr Elena: There is a genetic component, and I've actually seen this with a family that has a history of very problematic digestion of fat, and so on. Absolutely. But yeah, again, not all fats are made the same. And when you cut off the bad fats from your life, things change and everything changes really. Lisa: Yeah, it really is very satiating, too to have a little bit of fat and that can really help with cravings and blood sugar spikes—we're getting off topic.  So you have a company, NMN Bio, which produces nicotinamide mononucleotide supplement. And you've got a whole range of other stuff coming as well. Why did you decide like, you need to get this out there on the market? Based on your research and your knowledge around this area, why is it important that people take NMN if they're serious about slowing aging? Dr Elena: So first of all, I came across the biology of NAD and NMN during my PhD studies and my research kind of led me into this field because I was studying autophagy neurodegeneration. And actually, I still cannot disclose my research.  Lisa: Yeah, it’s not published yet.  Dr Elena: My research paper from my PhD is not published yet, but hopefully soon, so we're about to submit it quite soon actually. So for this reason, I started studying the biology of NAD and I actually saw how important and how crucial NAD is to the cell and what happens when we have a lack of NAD and depletion of NAD pools in the cell. And I've been supplementing with different kinds of vitamins and supplements my whole life really. So, I was watching closely this space for a while, and I was taking different supplements myself for a while.  And so, when I came across NMN and I realised that actually there is this strategy where we can supplement with a precursor in order to increase our energy levels, I found it really, really interesting. And I thought to give it a go myself and try it out and see the results. And then what shocked me was that the immediate effect of the supplement—so within a few days, you can already feel a difference in your energy levels and your focus. And this comes from the fact that sirtuins are responsible for so many molecular processes in the cell. And this is why you have this effect, including the mitochondrial biogenesis, which gives you basically increased ATP, consequently.  Lisa: You get actually more mitochondria. So, like, if you got heart disease...  Dr Elena: The production of more mitochondria, and then they produce more ATP as a result. And then you have this magic energy, yeah. This is why I thought to bring this product into the market. And the other reason was that there was not enough reliable suppliers on the market, which is crazy, because it's actually quite a popular supplement. It's been on the rise, the interest was rising for the past couple of years, but what we're seeing is there is a lot of white labelling companies that don't offer any certificates of analysis and so on. And also, you have even big companies not offering proper certificates of analysis, which was me like, it was…  Yeah, I don't understand. You have a big company, and you have just the purity report from like, 18 months ago, and you don't have any other analysis, such as heavy metals, or pH or microorganisms. So, the consumer is actually not confident in buying from you. And I wanted to deliver the best quality for myself and my family. And then I said, ‘Wait a minute. This is not done, right’. And this is why I launched the company because I wanted a company that was completely transparent. And I even say it on the website, that if you're interested in finding out who our suppliers are, and so on, and have any questions about our supply chain, just feel free to reach out to me. And I would be happy to disclose all of those things. There are other companies that you can't find any registration number, or who the founder is, and so on. And it's quite confusing, really, because like you— you don't know who you deal with.  Lisa: This is the same with the whole supplement industry. On the one hand, it's good that it's not regulated by the FDA, and whoever else, there are authorities around the world. Because like, then—they are turned into the pharmaceutical industry, which don't get me started. But on the other hand, there's not enough regulation around the quality control.  And one of the things when I was searching for NMNs, searching the world for it, I had to go overseas and import it to friends in America and get it out of there. And this is why I like—was super excited to discover your work. And then, we've since now made it available down here. So, we're going to branch down here in New Zealand for New Zealand, Australia. And I wanted someone who I could trust, who has all the scientific knowledge behind it, there's all lab tests, etc. And that was really important for me for quality.  Just on a side note. So I've been taking NMN now for—I think—so five, close to six months. I've had a massive weight loss and so as my mum. Why would that be? Like, I didn't take it for weight loss. I wasn't overweight, per se. But I had a couple of kilos that I was quite clear to get rid of. And what I've noticed—because I'm an athlete, that's my background—I haven't lost an ounce of muscle, which has been really awesome because most people are struggling to keep muscle mass, lose fat mass. My mum has lost 11 kilos. And she is of a genetic body type that really struggles with weight loss. She's conservation metabolism, from a genetic point of view, very, very hard for her to lose weight. So, I've never seen this in the history of her entire life, since I've been around. The weights just dropped off her.  Is this some sort of upregulation in the metabolic pathways? Is it improving the insulin resistance? What's it doing there to cause such weight loss without muscle loss? Dr Elena: Well, in my study so far, there's definitely evidence that it does improve insulin sensitivity, and it also improves the lipid metabolism profile. So those two are very important. And unfortunately, we don't have those studies in humans yet. But more clinical studies are on the way, and hopefully we'll have very good results this year with the NMN besides the safety studies that we already have in humans.  So in mice, what we're seeing is that there is basically a reverse of type two diabetes, which is really impressive. And if you want to correlate this data into humans somehow, I would say that, obviously, I'm not a medical doctor, and this is not a medical advice, but I would say that it does have to do something with the metabolism, and it basically improves the way your body metabolises everything. And...  Lisa: Worth trying and there's no downside to NMN. There's no, it's a vitamin B derivative, well then you will say to me, ‘Well, can I just take B3 and be done’? and it's like, no, it doesn't work like that, which should be a lot cheaper. Dr Elena: That’s the other impressive thing about this compound is that it actually doesn't have, if any, side effects at all. So even in studies with mice, where the dosage that they use in mice is actually much higher than it is in the one that we usually have in humans. So, for instance, if someone would take 500 mg, or one gram of NMN per day in humans. And then in mice studies, they use something like 200 mg per kilogram of weight, which is much, much more, and it still doesn't have any side effects. Lisa: Does it mean that we need higher dosages? Like in the human, or has it only been tested to one gram and why has it not been tested higher, if that's the case?  Dr Elena: No, I think that there are studies underway for this as well. So eventually, we will find what is the ideal dosage for humans. I think that from anecdotal evidence, people can already see results from 500 mg or one gram and so on. There are people that take more. So, some biohackers say that they take two grams or four grams, and is still very well tolerated. But yeah, so far, it does not produce any side effects in terms of… Lisa: Any downside. Dr Elena: Basically. And, for instance, for myself, my stomach is quite sensitive. So, when I'm on an empty stomach, I can't take vitamin C or caffeine and I get nauseous and so on. And this is not the case with an NMN. So, I can take it first. It is very well tolerated on an empty stomach, very mild. I really love it. There’s so many reasons to love it.  Lisa: Yeah, yeah, yeah, I have my morning and night. So, I'm on a gram a day. And is there any reason not to take it at night? So I split the dose—reasoning, thinking, keeping the levels up?  Dr Elena: I mean, I would probably take it all in the morning, I think. There's been a study out that it can affect the circadian rhythms as well. And interestingly, it actually affects NMN—sorry—NAD levels affects the circadian rhythm. But it's not the other way around. So, NAD actually dictates the circadian rhythm in the body. So, for this reason, I would suggest to take it in the morning because then your whole body synchronise, then you wake up and you tell to your body that look, it's the morning now, and we're going to have increased NAD level. Lisa: Increase. Ohh okay. So, okay, I got that wrong. I haven't noticed that I've had worse sleep or anything like that, or any rhythm has been out. But I would definitely swap to doing—my thinking process around that was keeping the tissue saturated over a 24-hour period, as opposed to all at once and then perhaps dropping, but I don't know. What is the half-life of it? Do you know? Is there any sort of evidence around that? Dr Elena: I actually, not sure. No, no.  Lisa: There’s no evidence yet. And so yeah, there's a ton of studies still being done that are currently, like this year, like going to be coming out, which is going to be really exciting. So that we're going to get more evidence. I mean, there's this stuff that I've been reading around fertility in animal studies, and they're starting to do human studies, which I personally am very interested in, in reversing aging of the ovaries and even with... I mean, the mice study was incredible around fertility, where the mice were postmenopausal, they actually knocked off any existing eggs with chemotherapy. And then gave them NMN and the mice went on to have babies. And there was a whole study. Dr Elena: This is why I get so excited about NMN and this is why it's my first product because frankly speaking as a scientist, I've never seen results like that with a natural compound.  Lisa: No? Dr Elena: Because there is a bunch of natural compounds out there, there is a bunch of other supplements. And what we're talking about spermidine the other day...  Lisa: Yeah, yeah, it's interesting.  Dr Elena: ...another autophagy activator. Quite an interesting supplement, yes. By the way, it's also an mTOR, independent autophagy activator, which is good. Lisa: Another very good reason to take that as well. And we were looking into that aren’t we, Elena about adding that?  Dr Elena: Yeah, absolutely. We will look into this, but again, you don't see results, like the ones that you see with NMN in multiple studies from other compounds, it's really fascinating. Lisa: Wow, so yeah, so there are other products that are going to... And this is a super exciting thing, like were our grandparents or our parents even didn't get the chance, like, with aging was aging, and there was nothing that you really could do to influence how fast you aged. They weren't aware of it. And later on, it's become well, if you eat better and you exercise a little bit more and you stop smoking and, and stuff, you’ll age slower. But now we're taking exponential leaps in our knowledge. I mean, I fell into this realm when I was reading Dr David Sinclair's book, who is a very prominent scientist at Harvard Medical School, and made his book, Lifespan, which I totally recommend people reading. I was just like, ‘Oh my gosh, if I can stay healthy now’, because I'm 52, ‘if I can stay like, really, in top shape for another 10 years, by then we're going to have stuff that will help me live really long’. And that really excites me. And not just live long, but live healthier.  Dr Elena: That’s the important part. The important part is not to just increase your age, it’s to increase your health span. So, the time that you're spending being healthy. And what you're referring to is actually called the aging escape velocity, where basically we’ll have more advanced research coming in every year of our lives. And this will eventually expand our lifespan, which is amazing. And I also think that if we preserve ourselves well, we might as well see this in our lifetimes, which will be amazing.  Lisa: Absolutely. And I want another few decades, please. Listening to Dave Asprey, who by far, got... Dr Elena: I want another like, few hundreds.  Lisa: Yeah, well, I mean, I know it sounds ridiculous right now. But if you listen to Dave Asprey saying conservatively, and Dr. David Sinclair, too, like, conservatively, we could live to 150, 180, and beyond. Then once they crack the code, and they're actually able to turn the cells back to which they are working on right now. And which they can actually go in the petri dish, from what I understand like with skin cells and make them immortal. And they can't do it in humans because it's too risky, they could turn you into a tumour and stuff. But with the Yamanaka factors that were discovered a decade or so ago, they're actually able to turn the clock back to the point of you being a 20-year-old again. And this is like, ‘Wow, this is pretty exciting. Being able to regrow nerves, spinal injuries, people who have gone blind from macular degeneration’ — all of these things are coming down the line. This is very, very exciting.  Dr Elena: There are several advancements in this field. So, as I said, my PhD is also in stem cell biology. So, I was working with human embryonic stem cells in the lab, and what they can do on a dish is just mind blowing. Because what I was able to do was to take human embryonic stem cells, and then dictate their fate, basically, with different growth factors, and then differentiate them into neural precursors at first. And then to push them further in order to become terminally differentiated neurons. And like four weeks later, you basically have a human brain in a dish and it's a primary human cells. And it's an amazing, physiological irrelevant human platform as well to study disease. And this is what I was doing during my PhD.  So, I’ve seen it with my own eyes. And every time I would do, I would go through this process, I would differentiate the human embryonic stem cells into neurons. It would be as exciting as the first time because of what it represents, because it does represent the progress that we've made so far. And I personally started human embryonic stem cells for the sake of drug discovery. So, I wasn't interested—my project was not focusing on different therapeutic applications. However, I know that there are many advancements in this field as well. So, we do have clinics in America, where you can have a total body rejuvenation, stem cells, and so on. And this technology is definitely advancing.  And I've been actually thinking about the application of this for myself. So as you know, I recently had a dental injury. This is something to keep in mind for the future. So perhaps in the near future, I can just inject myself with a bit of a stem cells there...  Lisa: And that’s already happening to a degree. I mean, I've got a doctor friend up north, who's doing stem cell replacement for joints, and so on, for degenerative joints. Because stem cells, basically, for people who don't understand why this is important. The stem cell is the original like cell, but before it decides, ‘Am I going to become a skin cell, or a neuron or a liver cell’, it differentiates. So, it's a pluripotent stem cell, it can become anything. And so, in the lab setting, you're going to be able to say, ‘Well I want your cell to become a liver cell’. Will we eventually be able to grow organs that can be used for transplantation? Is that sort of one of the end goals?  Dr Elena: Absolutely. And it's already been done with some organs. So for instance, I've heard that there is a research group that basically 3D-printed a functional thyroid gland from stem cells.  Lisa: Wow. 3D-printed. So, the printer gets these differentiated cells somehow, and then makes it into a functioning organ that they will eventually—they're going to be able to actually transplant this into people and save the whole organ donation, horrific troubles that we have currently.  Dr Elena: Yeah, exactly and I think that we're not too far away from this from whole organs being recreated in the lab. We already are able to actually do a 3D culture in the lab and create the so called organoids. So for instance, from stem cells, you can do a brain organoid, where you have a liposphere and it basically consists of different kinds of cells that you see in the brain. So it would have neurons, it would have glial, it will have astrocytes, and then it would have this brain organoid and then you can study it.  So, we're already getting there. We’re close, we’re much closer than we thought we were 20 years ago. And I think that we're not far away from having different kinds of organs being grown in the lab for transplants and so on.  Lisa: Hopefully not our brains because it's the seed of who we are. Honestly reading Dr Sinclair's book, I was like, ‘Am I in a Star Trek movie or something’? because it is pretty, pretty amazing. But when you do this, you also ask that to understand the whole process and how the whole thing functions, and then you can actually really slow down neurodegeneration and optimise things.  And so the NMN that we're talking about right now is the beginning of this really exciting road, which we're going to be staying abreast of. And hopefully adding to what we have available to the consumer right now for prices that are not moon money, that it's out of anybody's reach, but actually what you can do today so that you can preserve your health. So that in 10 years’ time, when the real crazy stuff starts coming on line, you'll be able to live longer and healthier lives. And that's the whole goal of it.  So before we just wrap up, I just wanted to reiterate again, so how is autophagy—can you just put that—how is autophagy related to NAD and sirtuin genes? Can you just put that two pieces together again, just repeat that a little bit? Dr Elena: Sure. So basically, what happens is that you do need autophagy to recycle different damaged organelles in the cell when something goes wrong. So, and this is quite prominent in neurodegeneration because the reason we have—let's say, aggregate from proteins in neurons and dysfunctional mitochondria and so on is because neurons are terminally differentiated cells. This means that they don't divide anymore. So, they rely on autophagy in order to have their housekeeping function because they can't divide the junk away. Okay. So that's the reason why autophagy is important in terminally differentiated cells such as neurons.  Lisa: So there's no hay flick limit for a neuron. There is just only one—when a neuron becomes a neuron, that's a neuron. Okay. Dr Elena: Yeah, yeah. And then that's it. And what happens with the activation of autophagy, one of the signals is—comes through sirtuin 1, which basically can activate the transcription factors that are related to autophagy activation, which is the TFEB transcription factor, EB and FOXO, which are basically influenced the activation of autophagy. And more specifically, the mitophagy as well. So, mitophagy is the arm of autophagy that is responsible for the mitochondrial clearance in the cell. Lisa: Yep, so mitochondria, just for people, are the powerhouses of the cell. This is where a lot of—so all of the energy is produced, if you like. And so, this is why mitophagy, as opposed to autophagy, so mitophagy is doing the same process, but within the mitochondria to keep your mitochondria healthy. And if your mitochondria are not healthy, and they're dying, and you're not having enough mitochondria in your cells, then you are going to be sick. And that could be heart disease, it could be neurodegeneration, that could be anything. So, keeping your mitochondria healthy is the basis of all bloody disease, blatantly.  Dr Elena: Yeah, exactly. So then, if you have impaired autophagy in the cell, and then you also have some sort of DNA damage going on, such as the one from reactive oxygen species, for example. And then what you have is the activation of the PARP enzymes. And PARP enzymes heavily rely on NAD levels in the cell in order to function. And NAD is also a substrate for the sirtuin genes that are responsible for also regulating a bunch of very healthy, a bunch of processes in the healthy cell. And for this reason, if you do have increased activation of PARPS, you will eventually get this NAD drain out of the cell. And this will not be enough in order for the sirtuins to function properly. And this will also deplete your autophagy. So, both NAD levels and autophagy are important to the cell. And fortunately for us, we can actually replenish the levels of NAD by supplementing with an ad precursor such as an NMN. Lisa: Okay, and so NMN has been proven to be by most of our bio available, because there's also like nicotinamide riboside which is used in a number of supplement companies that I know have nicotinamide riboside, but not many, there are some now, but have nicotinamide mononucleotide. Nicotinamide riboside is also a great molecule, but it's two steps away from becoming NAD. As long as it’s available.  Dr Elena: Yeah, so nicotinamide riboside needs to be phosphorylated and fast converted to nicotinamide mononucleotide first. And then this will enter the cell and then this will increase the levels of NAD in the cell. And for this reason—so first, this area of research was focusing on the NR molecule, the nicotinamide riboside. But then when they started studying NMN, they actually saw that there is increased bioavailability and there is increased levels of energy that come after supplementation with NMN. Lisa: Can you take—because NAD is a molecule, you cannot just take it as a capsule, and then it's all good to go. Can you take it as an infusion because I have heard of NAD infusions. I mean, it’s not available here.  Dr Elena: Well, and I'm curious myself about this, and I haven't done it, I haven't tested it. And from what I've seen—so the concentration of NAD in those intravenous injections is quite low. And I think that the same way that we have many opportunistic companies in the supplement field, we also have many opportunistic clinics that offer this kind of treatments. So, again, this is not something that I have studied in depth, and I actually don't know how much will it help. But yeah, I mean, this is another way to boost NAD, I guess, and you can try it out.  But with oral administration of NMN, we do have evidence that it can boost the levels of NAD in the tissue and in liver tissue and muscle tissue, and so on. And also, it's much easier to do and it's obviously much cheaper because those injections cost a lot. Lisa: Yes, yeah. Just one last question in relation to antioxidants, because I mean 10 years ago or so we used to think our reactive oxygen species ,oxidative stress happens through the electron transport chain. When we're metabolising, and so on, we get all these oxidative stresses and free radicals running around. And if we take antioxidants, we're going to be counterbalancing that. Does supplementing with antioxidants, like vitamin D, like glutathione, like vitamin C, and so on, alpha lipoic acid, is that going to contribute, too, to the slowing of aging, because it's going to down regulate the PARP enzymes? Dr Elena: People were very optimistic about antioxidants, something like 20 years ago. And everyone was talking about it and so on. But actually, the big studies that have been done, have shown that by taking antioxidants, you actually do not suppress aging. And there are some biomarkers that might have changed in those studies. But most of the biomarkers that they measure stay the same. Basically, saying that antioxidant is not the... Lisa: Not the holy grail. Dr Elena: ...that everyone was thinking about.  Lisa: Was hoping, yeah. Not to say that antioxidants don't have their place because they definitely do. Especially if you have a lot of oxidative stress, and you need to, like with vitamin C, if you're infected, or—I've done a whole series on vitamin C. But then it's not the holy grail for stopping the aging process, but it probably does help with not having so much PARP activation. I don't know, as a non-scientific brain, I'm just connecting dots.  Okay, so I think it's probably we've— so from a lifestyle intervention, apart from taking NMN and resveratrol, and oleic acid or olive oil, intermittent fasting, is there anything else that we can add to our anti-aging regime on a lifestyle intervention side?  Dr Elena: Intermittent fasting, and then avoid exposure to sunlight, as we said. And sirtuin genes are being activated from any kind of stress. And what we can do is we can also induce some sort of an artificial stress, which could be done, let's say with cryotherapy. This is what cryotherapy does. When you're exposed to cold, you also have this stress signal that activates sirtuins, or the other way around, so you can try out a sauna. And this will also have the same effect. So, I think this is also something to keep in mind.  Lisa: Breathing, breathing. So, sort of tumour breathing, or, like what one half does all of that sort of stuff. So, there’s hormetic stressors, there’s exercise obviously, that cause a cascade of changes and make you stronger. And yeah, it's sort of a balancing act. You don't want to be doing exercise for Africa or really freezing yourself to death, but you just want to have a little stress to cause a change in the body. So these hormetic stressors can be very, very helpful.  Okay, well, I think we've covered a very, very, very complex topic and I hope we didn't lose everybody on the way. But at the end of the day, take NMN, take resveratrol, take olive oil, do your exercise, get in the sauna, if you have a chance to do cold therapy, do that as well. Get your exercise, get your antioxidants in there as well, to a certain degree and you're going to be able to live long enough but until other things come online, and you'll be able to improve everything.  Dr Elena: Sounds good.  Lisa: Brilliant. So Dr. Elena, thank you very much. Dr. Elena has been on the show, NMN Bio. So we have nmnbio.co.uk in UK and in Europe, and nmnbio.nz if you're down at this end of the world. We'd love to help you over the air. If you've got any other questions, please reach out to us. And thanks very much for being here today. It's been really exciting. Dr Elena: Thank you, Lisa, thank you so much for having me. That's it this week for Pushing the Limits. Be sure to

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.18.303578v1?rss=1 Authors: Khayachi, A., Ase, A. R., Liao, C., Kamesh, A., Kuhlmann, N., Schorova, L., Chaumette, B., Dion, P., Alda, M., Rouleau, G. A., Milnerwood, A. J. Abstract: Bipolar disorder (BD) is characterized by cyclical alternations between mania and depression, often comorbid with psychosis, and suicide. The mood stabilizer lithium, compared to other medications, is the most efficient treatment for prevention of manic and depressive episodes. The pathophysiology of BD, and lithium mode of action, are yet to be fully understood. Evidence suggests a change in the balance of excitatory/inhibitory activity, favouring excitation in BD. Here, we sought to establish a holistic appreciation of the neuronal consequences of lithium exposure in mouse cortical neurons and identify underlying mechanisms. We found that chronic (but not acute) lithium treatment significantly reduced intracellular calcium flux, specifically through the activation of the metabotropic glutamatergic receptor mGluR5. This was associated with altered phosphorylation of PKC and GSK3 kinases, reduced neuronal excitability, and several alterations to synapse function. Consequently, lithium treatment shifts the excitatory/inhibitory balance in the network toward inhibition. Together, the results revealed how lithium dampens neuronal excitability and glutamatergic network activity, which are predicted to be overactive in the manic phase of BD. Our working model of lithium action enables the development of targeted strategies to restore the balance of overactive networks, mimicking the therapeutic benefits of lithium, but with reduced toxicity. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.19.258236v1?rss=1 Authors: Stojakovic, A., Chang, S.-Y., Nesbitt, J., Pichurin, N. P., Ostroot, M., Trushina, E. Abstract: Background: Accumulation of hyperphosphorylated Tau (pTau) protein is associated with synaptic dysfunction in Alzheimers disease (AD). We previously demonstrated that neuroprotection in familial mouse models of AD could be achieved by targeting mitochondria complex I (MCI) and activating the adaptive stress response. Efficacy of this strategy on pTau-related pathology remained unknown. Objective: To investigate the effect of specific MCI inhibitor tricyclic pyrone compound CP2 on pTau levels, memory function, long term potentiation (LTP), and energy homeostasis in 18-month-old 3xTg-AD mice and explore the potential mechanisms. Methods: CP2 was administered to male and female 3xTg-AD mice from 3.5 - 18 months of age. Cognitive function was assessed using the Morris water maze test. Glucose metabolism was measured in periphery using a glucose tolerance test and in the brain using fluorodeoxyglucose F18 positron-emission tomography (FDG-PET). LTP was evaluated using electrophysiology in the hippocampus. The expression of key proteins associated with neuroprotective mechanisms were assessed by western blotting. Results: Chronic CP2 treatment restored synaptic activity and cognitive function, increased levels of synaptic proteins, improved glucose metabolism and energy homeostasis in male and female 3xTg-AD mice. Significant reduction of human pTau in the brain was associated with increased activity of protein phosphatase of type 2A (PP2A), reduced activity of cyclin-dependent kinase 5 (CDK5) and glycogen synthase kinase 3{beta} (GSK3{beta}). Conclusion: CP2 treatment protected against synaptic dysfunction and memory impairment in symptomatic 3xTg-AD mice, and reduced levels of human pTau, indicating that targeting mitochondria with small molecule specific MCI inhibitors represents a promising strategy for AD. Copy rights belong to original authors. Visit the link for more info

Volume 11, Issue 25 of @Oncotarget reported that to examine the role of RSK in AML, the authors analyzed apoptosis and the cell cycle profile following treatment with BI-D1870, a potent inhibitor of RSK. BI-D1870 treatment increased the G2/M population and induced apoptosis in Acute Myeloid Leukemia cell lines and patient Acute Myeloid Leukemia cells. Therefore, the authors investigated whether BI-D1870 potentiates the anti-leukemic activity of vincristine by targeting mitotic exit. Combination treatment of BI-D1870 and vincristine synergistically increased mitotic arrest and apoptosis in acute leukemia cells. These data show that BI-D1870 induces apoptosis of AML cells alone and in combination with vincristine through blocking mitotic exit, providing a novel approach to overcoming vincristine resistance in AML cells. Dr. Kathleen M. Sakamoto from Stanford University School of Medicine said, "Acute myeloid leukemia (AML) is a genetically and phenotypically heterogeneous hematologic malignancy characterized by the accumulation of immature myeloid blasts with resultant peripheral blood cytopenia." Treatment of cells with microtubule targeting agents, including paclitaxel and the vinca alkaloid vincristine, blocks the proper formation of the mitotic spindle through inhibition of microtubule dynamics, resulting in the prolonged mitotic arrest of cancer cells. MTAs-treated mitotic arrested cells may undergo apoptosis in mitosis, however, the rapid degradation of Cyclin B due to an insufficient SAC leads to the mitotic slippage into tetraploid G1 stage in resistant cells. Though vinca alkaloid microtubule-destabilizing compounds have shown clinical efficacy against various hematological malignancies and were included in combination chemotherapy of the VAPA study, they are not currently used in induction chemotherapy for AML due to their high toxicity against lymphoid cells and rapid degradation by myeloperoxidase in AML cells. In this study, they demonstrate that BI-D1870, a potent inhibitor of RSK, induces mitotic arrest, and apoptosis in AML cells without inhibiting CDC2 and CDC25C. Furthermore, BI-D1870 synergizes with vinca alkaloid vincristine in AML cells, suggesting that inhibition of mitotic exit with BI-D1870 could be a promising novel approach for AML therapy in combination with MTAs. The Sakamoto Research Team concluded in their Oncotarget Research Paper that BI-D1870 is a reversible pan-RSK inhibitor, showing > 500-fold higher activity for RSK than other AGC kinases. BI-D1870 also inhibits the activity of PLK1, Aurora-B, MELK, PIM3, MST2, and GSK3β at higher concentrations than for RSK. BI-D1870 and BRD7389 have been reported to inhibit proliferation and significantly increase the G2/M population in melanoma cells. BI-D1870 does not have proper physicochemical properties for clinical application. Future structure-activity relationships study for BI-D1870 is required to improve solubility and pharmacokinetic profiles for in vivo preclinical and clinical studies. Sign up for free Altmetric alerts about this article DOI - https://doi.org/10.18632/oncotarget.27630 Full text - https://www.oncotarget.com/article/27630/text/ Correspondence to - Kathleen M. Sakamoto - kmsakamo@stanford.edu Keywords - acute myeloid leukemia, BI-D1870, RSK, vincristine, spindle assembly checkpoint About Oncotarget Oncotarget is a weekly, peer-reviewed, open access biomedical journal covering research on all aspects of oncology. To learn more about Oncotarget, please visit https://www.oncotarget.com or connect with: Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/ Oncotarget is published by Impact Journals, LLC please visit http://www.ImpactJournals.com or connect with @ImpactJrnls Media Contact MEDIA@IMPACTJOURNALS.COM 18009220957x105

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.06.18.159665v1?rss=1 Authors: Saundh, S., Patnaik, D., Gagne, S., Bishop, J., Lipsit, S., Amat, S., Pujari, N., Krishnan, A. K., Bigsby, R., Murphy, M., Tsai, L.-H., Haggarty, S., Leung, A. K.-W. Abstract: Glycogen Synthase Kinase 3-beta (GSK3{beta}) is a critical regulator of several cellular pathways involved in neuroplasticity and is a potential target for neurotherapeutic development in the treatment of neuropsychiatric and neurodegenerative diseases. The majority of efforts to develop inhibitors of GSK3{beta} have been focused on developing small molecule inhibitors that compete with ATP through direct interaction with the ATP binding site. This strategy has presented selectivity challenges due to the evolutionary conservation of this domain within the kinome. The Disrupted in Schizophrenia (DISC1) protein, has previously been shown to bind and inhibit GSK3{beta} activity. Here, we report the characterization of a 44-mer peptide derived from human DISC1 (hDISCtide) that is sufficient to both bind and inhibit GSK3{beta} in a non-competitive mode that is distinct from classical ATP competitive inhibitors. Based on multiple independent biochemical and biophysical assays, we propose that hDISCtide interacts at two distinct regions of GSK3{beta}: an inhibitory region that partially overlaps with the binding site of FRATide, a well-known GSK3{beta} binding peptide, and a specific binding region that is unique to hDISCtide. Taken together, our findings present a novel avenue for developing a peptide-based selective inhibitor of GSK3{beta}. Copy rights belong to original authors. Visit the link for more info

Dr Carolyn Lam:                Welcome to Circulation on the Run, your weekly podcast summary and backstage pass to the Journal and its editors. I'm Dr Carolyn Lam, associate editor from the National Heart Center and Duke National University of Singapore. Dr Greg Hundley:             And I'm Greg Hundley, associate editor at Circulation and director of the Pauley Heart Center at VCU Health in Richmond, Virginia. Carolyn, have you ever wondered about instead of coding a stent, coding balloons with paclitaxel? Well, the feature article day is going to look at mortality assessments of paclitaxel-coated balloons in a meta-analysis from the ILLUMENATE clinical program, the three-year outcomes. Do you have a paper you want to start us off? Dr Carolyn Lam:                I sure do. First of all, we know that diabetes impairs atherosclerosis regression following cholesterol lowering in both humans and mice. Now in this process of plaque regression, what's the role of functional high density lipoprotein or HDL, which is typically low in patients with diabetes?                                                 Well, this first paper that I chose looks just at that and it's from Dr Fischer from New York University School of Medicine and colleagues, who aimed to test if raising functional HDL levels in diabetic mice prevents monocytosis, reduces the quantity and inflammation of plaque macrophages and enhances atherosclerosis regression following cholesterol lowering. So to do this, the authors used aortic arches containing plaques, which were developed in LDL receptor null mice, and these were transplanted into either wild type or diabetic wild type or diabetic mice transgenic for human APL lipid protein A1, which have elevated functional HDL. Dr Greg Hundley:             So Carolyn, what did they find in this interesting study? Dr Carolyn Lam:                Well, diabetic wild type mice had impaired atherosclerosis regression, which was normalized by raising HDL levels. The benefit was linked to suppressed hyperglycemia-driven myelopoiesis, monocytosis and neutrophilia. Increased HDL improved cholesterol efflux from bone marrow progenitors, suppressing their proliferation and monocyte neutrophil production capacity. ACL also suppressed the general recruitability monocytes to inflammatory sites and promoted plaque macrophage polarization to the M2 phenotype, which is an atherosclerosis resolving state. There was also a decrease in plaque neutrophil extracellular traps or nets, which are atherogenic and increased by diabetes. So raising apolipoprotein AI and functional levels of HDL promoted multiple favorable changes in the production of monocytes and neutrophils and in the inflammatory environment of atherosclerotic plaques in diabetic mice after cholesterol lowering. And this may represent a novel approach to reduce cardiovascular risk in patients with diabetes. Dr Greg Hundley:             Really interesting, Carolyn. Well, I'm going to talk to you a little bit about a large study in patients with valvular heart disease and it's a contemporary presentation and management study and it's from the Euro Observational Research Program Valvular Heart Disease II, Roman numeral two, survey. And the corresponding author is Professor Bernard Iung from Bichat Hospital. So the VHDII survey was designed by the Euro Observational Research Program of the European Society of Cardiology to analyze actual management of valvular heart disease and compare practice with guidelines.                                                 Now in short, patients with severe and native valvular heart disease or previous valvular intervention were enrolled prospectively across 28 countries over a three-month period in 2017. Indications for intervention were considered concordant if the intervention was performed or scheduled in symptomatic patients corresponding to class one recommendation specified in the 2012 ESC and in the 2014 American Heart Association American College of Cardiology valvular heart disease guidelines. Dr Carolyn Lam:                Wow. So what did they find, Greg? Dr Greg Hundley:             Okay, so there's 7,247 patients. 4,483 were hospitalized, and 2,764 were outpatients, and they were included across 222 centers. The median age was 71 years and 1,917 patients were over the age of 80, and 3,400 were women. Now, aortic stenosis was present in 2,000 plus patients, aortic regurgitation in 279, mitral stenosis and 234, mitral regurgitation in 1,114. And multiple left-sided valvular heart disease was present in 1,297, right-sided valvular heart disease in 143, and 2,028 patients had prior vascular intervention.                                                 So the decision for intervention was concordant with class one recommendations in symptomatic patients with severe single left-sided valvular heart disease in 79.4% of those with AS, 77% with aortic regurgitation, 68.5% for mitral stenosis, and 71% for primary MR. Valvular interventions were performed in 2,150 patients during the survey. Of them, 47.8% of the patients with single left-sided native valvular heart disease were in New York Heart Association class three or four, and transcatheter procedures were performed in 38.7% of the patients with AS and 16.7% of those with MR. Dr Carolyn Lam:                Wow, Greg. So what are the take home messages? That was a lot of numbers. Dr Greg Hundley:             Yep. Lots of data there. And so couple things. First, recommendations for interventions in symptomatic patients with severe valve disease are better applied today in this paper than in the previous European survey conducted in 2001, particularly for those individuals with aortic valve disease. Second, multi-modality imaging is now more frequently used, but stress testing remains underused in asymptomatic patients. And finally, transcatheter therapies are now widely used in patients with stenotic valve disease, and we would expect that, particularly for the use in the elderly. Dr Carolyn Lam:                Great, Greg. So what are the clinical implications? Dr Greg Hundley:             Okay, so Carolyn, first, late referral for intervention shows the need for increasing awareness of valvular heart disease by general practitioners and cardiologists. Second, the high burden of elderly patients highlights the need for multidisciplinary heart team approaches to assess the risk benefit ratios of the different modalities of valvular interventions. And finally, number three, echocardiographic quantification of regurgitation should be more accurate and pay more attention to quantitative measurements. Those are the main take homes from this large registry analysis. Dr Carolyn Lam:                Nice. Thanks, Greg. My next paper is the characterization of the first transgenic mouse model of ARVC 5. Now, that is the most aggressive form of arrhythmogenic right ventricular cardiomyopathy caused by a specific mutation in transmembrane protein 43. So this paper's from co-corresponding authors, Dr Lara-Pezzi from CNIC in Madrid and Dr Garcia-Pavia from Hospital Universitario Porto de Hero in Madrid, and with their colleagues, they generated transgenic mice over expressing transmembrane protein 43 in either it's wild type or that specific mutant form in postnatal cardiomyocytes under the control of alpha-myosin heavy chain promoter.                                                 And they found that these transgenic mice expressing the specific mutant in transmembrane protein 43 showed fibro fatty replacement of the myocardium and died at a young age. The model confirmed that transmembrane protein 43 is mostly localized at the nuclear membrane and provides new information regarding the pathophysiological mechanisms underlying ARVC five. One of them is that the GSK3 beta signaling pathway plays an important role in this disease. Dr Greg Hundley:             So that's great, Carolyn. Sounds like we have a new model that's been created by this group and certainly this disease has spread. It's something we definitely worry about. Do you see any therapeutic implications for their work? Dr Carolyn Lam:                Great question, and indeed the authors tested two new therapeutic approaches for ARVC five. In the first they found that targeting fibrosis really had no beneficial effect. But in the second, they found that inhibition of GSK3 beta improved cardiac function and survival, thus opening the way to a new therapeutic approach focused on GSK3 beta inhibition in patients with ARVC five. Dr Greg Hundley:             Very good. So we look forward to seeing what the results of that study will be. How about now we talk about some of the other articles in this issue? Dr Carolyn Lam:                I love that. I think it's a great idea to tell everybody about this amazing issue. So we start with an article from our Global Rounds, and this time from Argentina, so a great status update and future strategies for cardiovascular disease in Argentina. We also have a perspective paper and that's on the new World Symposium on Pulmonary Hypertension guidelines, really questioning some of the cutoffs that we've taken for granted and asking, "Should 21 be the new 25?" Intrigued? Well, you really need to pick this one up and read it.                                                 And then there's a white paper, and this is a report from the 2018 NHLBI workshop that really talks about unlocking the secrets of mitochondria in the cardiovascular system and asking if this may be a path to cure in heart failure. We also have a research letter, and I love these. They're so succinct and really contain an important message. And this one talks about the evolution of Medicare formulary coverage changes for antithrombotic therapy after the guideline update. So very topical subject. Dr Greg Hundley:             Very good, Carolyn. So I've got a couple. There's a Paths to Discovery article that John Rutherford did discussing with Paul Zimmet regarding reflections of the evolving global diabetes epidemic. Second, there is a very nice On My Mind piece from Samuel Tretheway from Birmingham, England who discusses medical misinformation, kind of like medical fake news. And he discusses how this occurs and it depends on the motivation of both authors and publishers, and he reviews responsibilities of all of us, how to avoid generating this type of material. And then finally, a really interesting Cardiology News piece by Bridget Kuehn, who discusses diet and microbes in heart failure, and with that there's a very nice piece of artistry work that would be great for your office. So that's all included in the journal. Dr Carolyn Lam:                Oh, you got us all curious. Finally, I just want to highlight, we have a section called Highlights from Major Meetings, and this time from my part of the world with Dr Aijun Sun and Dr Junbo Ge summarizing the 13th Oriental Congress of Cardiology takeaways. Cool issue, isn't it? Dr Greg Hundley:             Absolutely. So how about onto our feature discussion? Dr Carolyn Lam:                You bet, Greg. Dr Greg Hundley:             Welcome everyone to our feature discussion. And this afternoon or this morning, wherever you may be, we are going to have an opportunity to discuss the utility of paclitaxel-coated balloons in terms of management of patients with peripheral arterial disease. And our article today comes to us from Bill Gray and colleagues from Mainline Health in Philadelphia, Pennsylvania. And we have our own Josh Beckman, associate editor from Vanderbilt, who will be joining us in the discussion. Bill, welcome to Circulation. We really appreciate you sending us this article. Can you tell us a little bit about the background of why you wanted to perform your study and also, what was your study design, study population? Dr William Gray:               The study was really prompted by a prior report by Katsanos et al in JAHA about nine months ago. When we started this study, it was much more fresh. And what we did was we realized we had data from multiple studies using the Stellarex drug-coated balloon that we could use to address some of the issues raised with the Katsanos paper. Just to review that briefly, the Katsanos paper suggested that there was a significant mortality signal in patients who were randomized to drug-coated balloons using paclitaxel versus PTA or patients randomized to drug eluting stent versus PTA or other stents. That signal was seen late at two years and at five years, and so we sought a given the data, the tightly controlled and well-reported data and this experience to see if we could see a signal as well.                                                 The study design really involved taking all the data from the randomized trials, and there were two, which comprised an aggregate of about 600 patients, unequally randomized, about 400 in the drug-coated balloon arm and about 170 or 200 patients in the PTA arm. And then we also looked at all the poolable data, which was controlled data, so we had two randomized control studies I mentioned just a minute ago, as well as three single arm studies in one registry. Now, these had quality oversight and data reporting. And then those data were adjudicated for adverse events, including death, by a blinded third party CEC, and then those data reported out by Kaplan–Meier estimates as well, and then we do a multi-variable analysis looking at predictors of death, and then I can talk about that in a moment. Importantly, the data here has followed out to three years. As I mentioned before, the original paper which incited the concern had reported unequal deaths at two and five years, so we're somewhere splitting that difference. That's the genesis of the study and the study design. Dr Greg Hundley:             So Bill, tell us now about the results. Dr William Gray:               It turns out the baseline characteristics were largely similar between these trials and the patient arms, even though they weren't strictly speaking the same trials, except that the drug-coated balloon arm was a bit younger and smoked more frequently, so they were at a little bit more risk. In the randomized control analysis, which was done first, there was no difference in all-cause mortality between the PTA patients and the patients who received paclitaxel drug-coated balloons. That was true at one year, two years and three years. When we looked at the pooled analysis, which included not only the drug-coated balloon randomized trial patients, but also all the single arm studies and registries, we also found that there was no differences between those treated with drug-coated balloons in those additional studies and the control group of 170 patients in the randomized trial arm of PTA alone.                                                 Interestingly, when we started to look at the multi-variable analyses, we did something that we ordinarily would not do, but because of the pressing issue around paclitaxel mortality, we actually did a standard covariate analysis looking at predictors and then we forced drug and drug dose into the model to see if they would come up positive as a predictor of outcome. As you might expect, not surprisingly, we found that age, congestive heart failure, diabetes and renal insufficiency were the four major predictors of mortality in a group of patients who were largely claudicates with significant peripheral vascular disease. No surprise there. We all know the patients don't die of claudication, they die of cardiovascular disease, and this I think bears that out.                                                 When we force drug into the model, in point of fact, not a dose nor the presence of drug had any impact on death rates in the model, so there was no predictive value there whatsoever. Those are the results. Again, they're out to three years, and I think one of the important things that we have to recognize is that the numbers are relatively small and the follow-up is relatively limited and by itself, although it doesn't show any signal, it probably doesn't stand on its own to refute a larger meta-analysis, but does I think contribute to the dataset that is becoming more evident that the individual analysis do not appear to show mortality effects. Dr Greg Hundley:             Very good. So this is Dr Josh Beckman at Vanderbilt University. Josh, could you talk to us a little bit and put this paper in perspective relative to the prior published literature in terms of how you manage patients with peripheral arterial disease? Dr Joshua Beckman:        I have to say first, I'm really glad that we're able to publish this paper from Bill Gray and his group. We are, and I'm going to put this in really muted terms, in extraordinary times. I have never seen what is going on now happen with any other technology or really even medical therapy in the 20 plus years I've been a practicing physician. I think for the audience, it's really important to understand what is going on right now because if you don't pay attention to this space, you may not realize what's really been happening. Bill did a nice job at telling you why he did the study, which was this Katsanos aggregate level meta-analysis that was published in JAHA back in December.                                                 On the basis of this paper, there has been a rapid development of worry and concern that these devices may be associated with late mortality. This concern has spread to the Food and Drug Administration, which has now put out three letters to healthcare professionals, each of them basically suggesting that you should choose non drug-coated either balloons or stents first, and if you want to use these, you have to have an extended conversation with the patients discussing the risks. And so in response to this aggregate level meta-analysis, which had an extensive number of lost to follow-up patients and didn't account for crossovers and the usual problems with this kind of information, I have been really impressed by the community of people who are interested in this topic and work with these kinds of devices.                                                 And by that, I mean, the response has not just been a series of editorials. The response has really been, "Let's find every single piece of data that we can find to see whether or not this signal holds up," because as evidence-based physicians, we take one piece of data and say that it is one piece of data, and then we have to put it into the context of all of the other pieces of data that were published. And so I know that Dr Gray is old enough to remember 10 years ago when these devices were being used in the coronary arteries with drug eluting stents. And as far as anybody can tell with studies that were two to three times larger or meta analyses two to three times larger than the study published in December, there was no mortality signal.                                                 It should be made clear that in doses that dwarf the doses from these devices, when these medications are given to pregnant women who have breast cancer, not only is the mother fine but the fetus is fine. And so I think paper that we are discussing this morning in particular, but the group of investigators in the space has really stepped forward to publish as much data as possible to fill out our understanding and place the original study in the correct context. And so when you understand what's happening in the community, and there's been a significant reduction in the use of these devices on the basis of that one publication at the expense of patients for whom these devices are really much better at limb outcomes, then you can understand why we were so interested in the paper by Dr Gray.                                                 This is another brick in creating the foundation to really have a fuller and better understanding of any possible relationship between the use of these devices and a nonspecific increase in mortality two to five years later, which as far as I can tell, I've never seen something that may end up being a poison that doesn't have a specific mechanism of causing morbidity or mortality. And so when we got this paper, I was really happy to be able to work with Bill and bring it to the level that it is now so that when it's published in October, it's going to be another really important contribution and I just want to congratulate the authors for doing that work. I will say, and I'd like to get Bill's perspective on how he thinks the information that's now being published is going to help us understand what to do with these devices. Dr William Gray:               Yeah, that's a great question, and I want to emphasize something you brought up, which I did not, which is at the aggregate level data that Katsanos used to publish his analysis was really all he had access to, which means that he had some numerical data from prior published publications but did not have patient level data. And so what Josh is referring to appropriately is the concept that each individual holder of those data, those patient level data, are now coming forward with their own analysis of those data at a patient level, which allows us to look more granularly and more clearly at the causes of death. For example, in this study, the causes of death did not cluster around cancer. They were largely cardiovascular, and they were not dis-equally distributed or unequally distributed between the two groups.                                                 So I think that patient level data, to get back to your original question, Josh, the patient level data will be incredibly important from each of the experiences with the various drug-coated balloons and drug eluting stents on the market because it does allow us to look more closely at the mechanism of death and whether there's any putative cause that might be assigned to paclitaxel. As you mentioned, the pharmacology of this is not understandable. The only type of pharmacology that would work like this was if paclitaxel was radioactive and accumulated a hazard along the way, but we know that's not true.                                                 I think extend your question, it's important to say that both the FDA and other independent groups like VIVA have looked closely at the meta analytic data both from a patient level and aggregate level data set, and they have seen a signal at five years. The problem with that is that data starts to winnow down very quickly at five years. There's not a lot of numbers, so that's the first problem, and the meta-analysis that have followed the publication by Katsanos. The second problem is, as Josh alluded to, there's a lot of missing data. Either patients withdrew or got lost to follow-up, and that didn't happen at an equal distribution between the control and the active arms, so there's some ascertainment bias there.                                                 And lastly, there's a crossover, that is patients who are in the control arm crossed over near as we can tell at a rate of about one in five or one in four to an active arm in the first year alone, which means they need to be reassigned to a risk pool that includes the original assignment of paclitaxel randomization. My sense is that those data will not get any better in the near-term future because the problems I just listed are not going to go away anytime soon. And so we are left with these individual patient level data and other big data, like Medicare analyses of tens of thousands of patients or Optum insurance analyses of again, tens of thousands of patients, which actually show no difference between the treatment with paclitaxel in the real world and patients treated with non-paclitaxel devices. So while we are comfortable and happy to publish these data and we think that are meaningful in terms of contributing to the larger dataset, we recognize the flaws and the limitations in the meta-analysis, which will not be solved soon or quickly. Dr Joshua Beckman:        So, I totally agree with what you just said. I will also say that every time data like this is published, it adds to the picture to make our understanding clearer. And you are responding directly to the Food and Drug Administration, who basically said they are not settled on this question either. It is noted, they are worried about it, and what they've really asked for is for more data to be published. And so when people analyze data like these, I think it is really helpful to the rest of us to create a fuller and more granular picture of the overall state of the field. Dr Greg Hundley:             We want to thank again both Josh for his time and Bill for his time. Hope you have a great week, and both Carolyn and I look forward to sharing with you again next week. Take care everyone. Dr Carolyn Lam:                This program is copyright American Heart Association 2019.  

On the latest episode of the Greg Suess Podcast, Greg sits down with Karen Gamble, associate professor in Psychiatry at the University of Alabama Birmingham and a GSU Brains and Behavior alum. They begin with reminiscing about a GSU homecoming and inspirations for going into science (1:48), the life of electrophysiology experiments during post-doctoral training (10:10), the role of GSK3 in the modulation of the molecular clock (14:53), the properties of SCN neurons (27:02), how circadian work can be used to study shift work (28:11), how chronobiology research can help understand human disorders (38:03), reminiscing about graduate school days at GSU in Atlanta (40:38), the exciting science at UAB and the Neuroscience Roadmaps Scholars program (42:53), and how Birmingham is as a city (51:59).

V tomto podcaste budeme hovoriť o výsledkoch štúdie, ktorá naznačuje, že možno by sme sa mali trošku krotiť s vŕtaním zubov a že možno existujú aj menej invazívne spôsoby, ako si poradiť so zubným kazom. YouTube TémyZdroje Intro Zubné plomby Outro Australian study finds 'no-drill' dentistry stops tooth decay Promotion of natural tooth repair by small molecule GSK3 antagonists

FK506 binding protein 5 (FKBP5) has been linked to stress related diseases and treatment response in depression (Binder et al., 2004). The corresponding protein FKBP51 was first identified as co-chaperone of HSP90 in a complex with steroid hormone receptors, where it diminishes hormone affinity and nuclear translocation efficiency of the receptors (Pratt and Toft, 1997; Wochnik et al., 2005). With FKBP5 transcription being induced by glucocorticoid signalling, an ultra-short feedback loop is provided for regulation and termination of GR activity. Dysregulation of this ultra-short feedback loop interferes with the stress hormone regulation and likely contributes to the association of FKBP5 with stress-related psychiatric disorders. Recently, important actions of FKBP51 beyond glucocorticoid signalling have been characterised in shaping the posttranslational regulation of certain molecular pathways in response to treatment with particular psychopharmaca (Gassen et al., 2014, 2015). As a contribution to elucidating the role of FKBP5 in stress related diseases, a two-sided approach was taken in this study by analysing the role of FKBP5 in regulation of transcription and in calibrating the responsiveness of these pathways to psychopharmacological treatment. To elucidate the transcriptional effects of FKBP5 in an unbiased approach, the expression profile of mice with deleted FKBP5 and their litter mates with functional FKBP5 were compared. A marked difference in glyoxalase-1 (GLO1) transcription was observed with higher GLO1 transcription in mice with deleted FKBP5, which was reflected by about two-fold more GLO1 protein in these mice. The efforts in deciphering the role of FKBP5 in elevation of GLO1 expression led to the identification of a duplication of the GLO1 gene inherent to mice with deleted FKBP5; this likely explains the enhanced GLO1 expression in these mice. This observation exemplifies the flanking gene problem and is a note of caution for interpreting data from conventionally generated knock-out mice. Overall, deletion of FKBP5 did not markedly change gene expression. In the second part of this thesis, the molecular effects of psychopharmacologic drugs were profiled for their dependency on FKBP51 function to modulate intracellular pathways relevant for treatment outcome in a cellular FKBP5 knockout model. For this purpose, psychopharmaca from the classes of SSRIs, SSNRIs, TCAs, atypical antidepressants, mood stabilisers, and NMDA receptor antagonists were analysed. In addition to GSK3β and AKT, which were reported to interact with and be targeted by FKBP51 recently (Gassen et al., 2015; Pei et al., 2009), ERK was identified as a novel kinase interacting with and being targeted by FKBP51 in this work. With GSK3β, AKT, and ERK, three major kinases were observed to be regulated by psychopharmaca. The effects were not homogeneous across all psychopharmaca and only loosely followed drug classes. Moreover, regulation of these kinases as well as their downstream targets was non-uniformly influenced by FKBP51. With FKBP51 being a stress induced gene, this transcriptional mechanism efficiently links the stress response to the regulation of the targets analysed in this work. Moreover, markers of autophagy, a cellular degradation process which has been linked to neurotransmission, were detected to be regulated by valproic acid (VPA), a mood stabiliser with HDAC inhibitory activity. VPA, as well as a second HDAC inhibitor butyric acid (BUT) enhanced the transcription of late and delayed autophagy markers controlled by FOXO3 signalling. Considering the versatile action of FKBP51 on targets analysed in this work, the list of proteins modulated by FKBP5 seems by far not complete. The diversity of effects evoked by different psychopharmaca hints to superimposed molecular effects underlying treatment outcome. Better understanding of pathway responsiveness could yield molecular markers for personalised medication that could be utilised to improve treatment outcome in stress related psychiatric diseases.

We have investigated molecular mechanisms for muscle mass accretion in a non-inbred mouse model (DU6P mice) characterized by extreme muscle mass. This extreme muscle mass was developed during 138 generations of phenotype selection for high protein content. Due to the repeated trait selection a complex setting of different mechanisms was expected to be enriched during the selection experiment. In muscle from 29-week female DU6P mice we have identified robust increases of protein kinase B activation (AKT, Ser-473, up to 2-fold) if compared to 11- and 54-week DU6P mice or controls. While a number of accepted effectors of AKT activation, including IGF-I, IGF-II, insulin/IGF-receptor, myostatin or integrin-linked kinase (ILK), were not correlated with this increase, phosphatase and tensin homologue deleted on chromosome 10 (PTEN) was down-regulated in 29-week female DU6P mice. In addition, higher levels of PTEN phosphorylation were found identifying a second mechanism of PTEN inhibition. Inhibition of PTEN and activation of AKT correlated with specific activation of p70S6 kinase and ribosomal protein S6, reduced phosphorylation of eukaryotic initiation factor 2α (eIF2α) and higher rates of protein synthesis in 29-week female DU6P mice. On the other hand, AKT activation also translated into specific inactivation of glycogen synthase kinase 3ß (GSK3ß) and an increase of muscular glycogen. In muscles from 29-week female DU6P mice a significant increase of protein/DNA was identified, which was not due to a reduction of protein breakdown or to specific increases of translation initiation. Instead our data support the conclusion that a higher rate of protein translation is contributing to the higher muscle mass in mid-aged female DU6P mice. Our results further reveal coevolution of high protein and high glycogen content during the selection experiment and identify PTEN as gate keeper for muscle mass in mid-aged female DU6P mice.

In this study a structure–function analysis has been employed to analyze transcriptional regulation through the Mediator subunit MED25. A relationship could be established between predicted structural domains and functional characteristics of this protein. Most critically the region responsible for interaction of MED25 with the Mediator was identified. Immunoprecipitation experiments demonstrated that the so–called VWA domain (von–Willebrand A domain, amino acids 1–290) is both sufficient and required for this contact. Site–directed mutagenesis indicates that this binding reaction involves the non–conserved loop SR2, which is protruding from this domain. Based on the results of this analysis a model was proposed, in which the primary contact is established by ionic forces and is further stabilized by hydrophobic interactions. The previously identified ACID domain was reported to bind to VP16. Targeted mutagenesis of four different motifs in this region impaired not only transcriptional activation through MED25 but also led to reduced binding to VP16. In particluar a lysine–rich motif is also present in two domains of PTOV1, a close homolog of MED25. Noteworthy, K518 is not conserved in the PTOV1_B domain, which in contrast to PTOV1_A and the ACID domain of MED25 does not bind to VP16. This led to the hypothesis that K518 is critically involved in the binding of VP16 to MED25. Furthermore it could be demonstrated that MED25 contains an intrinsic transcriptional activation capacity, which is localized in the region 290–715. This indicates additional recruitment of other factors to promoters through this region. Together with the Mediator binding VWA–domain and the VP16–interaction domain this region might facilitate transcriptional activation. A genome–wide screen showed downregulation of c–Jun and FosB following overexpression of MED25. Interestingly, expression of GSK3β, a downstream target of which is cyclin D1, seems to be stimulated by MED25. Together with the finding that overexpression of MED25 leads to activation of a p21 reporter, this raises the possibility that MED25 is involved in cell cycle control. An overlap has been discovered by comparison of MED25 target genes and genes identified previously as target for the viral activator EBNA2. The close homology between the activation domains of EBNA2 and VP16 implies a common mechanism of transcriptional activation by these two viral proteins through MED25. The involvement of MED25 in gene activation by viral activators might indicate a role for this Mediator subunit in viral transcription.