Podcasts about rt qpcr

BUFFALO, NY — February 26, 2025 — A new #research paper was #published in Aging (Aging-US) on January 27, 2025, in Volume 17, Issue 1, titled “Age-invariant genes: multi-tissue identification and characterization of murine reference genes.” Aging is a process driven by changes in gene activity, but researchers from Yale University School of Medicine and Altos Labs, led by first author John T. González and corresponding author Albert T. Higgins-Chen, have identified a set of genes that remain unchanged throughout the aging process. This discovery could improve the accuracy of aging research and provide insights into why some genes stay unchanged while others decline. “Reference genes have mostly been identified and validated in young organisms, and no systematic investigation has been done across the lifespan.” The study looked at gene activity in 17 different tissues in mice, from 1 month old to over 21 months old. Scientists used advanced bioinformatic analysis methods to analyze RNA sequencing data. They found nine genes that stayed the same across all tissues, as well as other genes that remained stable in specific tissues. These genes are usually shorter and have special DNA regions called CpG islands, which may help cells stay healthy and resist aging. Their stability throughout aging was confirmed by analyzing different datasets and using RT-qPCR. One of the most significant findings is that these stable genes are linked to essential cellular functions, such as mitochondrial activity and protein maintenance. This challenges the common belief that all aspects of aging involve gene dysregulation. Instead, the findings suggest that some cellular processes may naturally resist aging, leading the way for new research on longevity and potential anti-aging therapies. “Biological processes that change with age and those that resist age-related dysregulation are two sides of the same coin, and both will need to be investigated to fully understand aging.” Another key finding is that commonly used reference genes, such as GAPDH and ACTB, fluctuate with age, making them unreliable for aging studies. No single classical reference gene was found to be stable across all tissues. Researchers often use these reference genes as a control to measure gene activity, but if their expression changes over time, it can lead to inaccurate results. By identifying new, stable reference genes, this study provides scientists with better tools for studying aging-related diseases, regenerative medicine, and longevity science. Understanding how certain genes remain unchanged throughout life suggests that they may play a protective role in aging and could potentially be used to develop treatments that slow down age-related decline. While further research is needed, this discovery sets a new standard for measuring gene activity in aging studies and could have a significant impact on aging research and medicine. DOI - https://doi.org/10.18632/aging.206192 Corresponding author - Albert T. Higgins-Chen - a.higginschen@yale.edu About Aging-US The mission of the journal is to understand the mechanisms surrounding aging and age-related diseases, including cancer as the main cause of death in the modern aged population. The journal aims to promote 1) treatment of age-related diseases by slowing down aging, 2) validation of anti-aging drugs by treating age-related diseases, and 3) prevention of cancer by inhibiting aging. (Cancer and COVID-19 are age-related diseases.) Please visit our website at https://www.Aging-US.com​​ and connect with us: Facebook - https://www.facebook.com/AgingUS/ X - https://twitter.com/AgingJrnl Instagram - https://www.instagram.com/agingjrnl/ YouTube - https://www.youtube.com/@AgingJournal LinkedIn - https://www.linkedin.com/company/aging/ Pinterest - https://www.pinterest.com/AgingUS/ Spotify - https://open.spotify.com/show/1X4HQQgegjReaf6Mozn6Mc Media Contact 18009220957 MEDIA@IMPACTJOURNALS.COM

Featuring an interview with Dr Seth Wander, including the following topics: Therapy selection after CDK4/6 inhibitor failure: A review of current and investigational treatment for HR-positive, HER2-negative breast cancer Astore S et al. A therapeutic algorithm guiding subsequent therapy selection after CDK4/6 inhibitors' failure: A review of current and investigational treatment for HR+/Her2- breast cancer. Crit Rev Oncol Hematol 2024;204:104535. Abstract (0:00) A preoperative window-of-opportunity study of the oral SERD imlunestrant for newly diagnosed ER-positive, HER2-negative localized breast cancer Neven P et al. A preoperative window-of-opportunity study of oral SERD, imlunestrant, in newly diagnosed ER-positive, HER2-negative early breast cancer: Results from the EMBER-2 Study. Clin Cancer Res 2024;30(23):5304-13. Abstract (3:30) An assessment of an exosome-based ESR1-monitoring RT-qPCR kit that detects acquired resistance variants in liquid biopsy samples Statt S et al. An exosome-based ESR1 monitoring RT-qPCR kit that rapidly and accurately detects acquired resistance variants at ≤ 0.1% frequency in liquid biopsy samples. ESMO 2024;Abstract 420P. (7:08) CME information and select publications

Hello ! Aujourd'hui je vous retrouve dans cet épisode pour vous parler d'une technique fondamentale en biologie moléculaire : la RT-qPCR

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.26.538466v1?rss=1 Authors: Siddiq, M. M., Toro, C. A., Johnson, N. P., Hansen, J., Xiong, Y., Mellado, W., Tolentino, R. E., Johnson, K., Jayaraman, G., Suhail, Z., Harlow, L., Beaumont, K. G., Sebra, R. G., Willis, D. E., Cardozo, C. P., Iyengar, R. Abstract: Introduction- Neurons transport mRNA and translational machinery to axons for local translation. After spinal cord injury (SCI), de novo translation is assumed to enable neurorepair. Knowledge of the identity of axonal mRNAs that participate in neurorepair after SCI is limited. We sought to identify and understand how axonal RNAs play a role in axonal regeneration. Methods- We obtained preparations enriched in axonal mRNAs from control and SCI rats by digesting spinal cord tissue with cold-active protease (CAP). The digested samples were then centrifuged to obtain a supernatant that were then sequenced. We used bioinformatics analyses to identify DEGS and map them to various biological processes. We validated the DEGs by RT-qPCR and RNA-scope. Results- The supernatant fraction was highly enriched for axonal mRNA. Using Gene Ontology, the second most significant pathway for all differentially expressed genes (DEGs) was axonogenesis. Among the DEGs was Rims2, which is predominately a circular RNA (circRNA) in the CNS. We show that Rims2 RNA within spinal cord axons is circular. We found an additional 200 putative circRNAs in the axonal-enriched fraction. Knockdown in primary rat cortical neurons of the RNA editing enzyme ADAR1, which inhibits formation of circRNAs, significantly increased axonal outgrowth. Focusing on Rims2 we used Circular RNA Interactome to predict that several of the miRNAs that bind to circRims2 also bind to the 3 prime UTR of GAP-43, PTEN or CREB1, all known regulators of axonal outgrowth. Axonally-translated GAP-43 supports axonal elongation and we detect GAP-43 mRNA in the rat axons by RNAscope. Discussion- By using our method for enrichment of axonal RNA, we detect SCI induced DEGs, including circRNA such as Rims2. Ablation of ADAR1, the enzyme that regulates circRNA formation, promotes axonal outgrowth of cortical neurons. We developed a pathway model using Circular RNA Interactome that indicates that Rims2 through miRNAs can regulate the axonal translation GAP-43 a known regulator of axonal regeneration indicating that axonal mRNA contribute to regeneration. 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/2023.04.12.536647v1?rss=1 Authors: Hedley, K. E., Cuskelly, A., Quinn, R. K., Callister, R. J., Hodgson, D. M., Tadros, M. A. Abstract: Early-life inflammation can have long lasting impact on pain processing and pain behaviours. For example, we have shown neonatal inflammation can result in changes within spinal neuronal networks and altered flinching of the hind paw 5 following formalin injection three weeks later. This suggests mechanisms for altered pain behaviours lie in first and second order neurons in the pain neuroaxis. Exactly how these changes progress during postnatal development is not known. Accordingly, we investigated neuroinflammatory markers in sensory neurons (dorsal root ganglia; DRGs) and spinal cords of Wistar rats (both sexes) after 10 early life inflammation. Rats were injected with LPS or saline on postnatal days (P) 3 and 5. DRGs and spinal cords (SC) were isolated on P7, 13 and 21, and the expression of six inflammatory mediators were quantified via RT-qPCR. In the DRG, four proinflammatory mediators were elevated in P7 rats exposed to LPS. By P13, only two proinflammatory agents were elevated, whereas at P21 the levels of all six inflammatory mediators were 15 similar between LPS and saline-treated rats. There were no sex-specific differences in the expression profile of any mediator in DRGs. In the spinal cord this expression profile was reversed with no change in inflammatory mediators at P7, elevation of two at P13 and four at P21 in LPS treated rats. Interestingly, these differences were greater in the spinal cords of female rats, indicating sex-specific modulation of neuroinflammation even at these early 20 stages of postnatal development. The increased inflammatory mediator profile in the spinal cords of P21 LPS-treated rats was accompanied by sex-specific modulation of astrocytic (GFAP) activation, with females showing an increase and males a decrease in GFAP following LPS exposure. Together, these data indicate sensory neurons are more susceptible to acute inflammation whereas inflammation in the spinal cord is delayed. The sex-specific 25 modulation of inflammation during critical phases of development may help explain altered pain behaviours in adult males and females. 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/2023.04.12.536514v1?rss=1 Authors: Awatade, N. T., Reid, A. T., Nichol, K. S., Budden, K. F., Veerati, P. C., Pathinayake, P. S., Grainge, C. L., Hansbro, P. M., Wark, P. A. Abstract: Introduction: Primary air liquid interface (ALI) cultures of bronchial epithelial cells are used extensively to model airway responses. A recent advance is the development of conditional reprogramming that enhances proliferative capability. Several different media and protocols are utilized, yet even subtle differences may influence cellular responses. We compared the morphology and functional responses, including innate immune responses to rhinovirus infection in conditionally reprogrammed primary bronchial epithelial cells (pBECs) differentiated using two commonly used culture media. Methods: pBECs from healthy participants (n = 5) were CR using gamma-irradiated 3T3 fibroblasts and Rho Kinase inhibitor. CRpBECs were differentiated at ALI in either PneumaCultTM (PN-ALI) or Bronchial Epithelial Growth Medium (BEGM)-based differentiation media (BEBM:DMEM, 50:50, LonzaTM) - (AB-ALI) for 28 days. Transepithelial electrical resistance (TEER), immunofluorescence, histology, cilia activity, ion channel function, and expression of cell markers were analyzed. Viral load was assessed by RT-qPCR and anti-viral factors quantified by Legendplex following Rhinovirus-A1b (RVA1b) infection. Results: CRpBECs differentiated in PneumaCultTM were smaller and had a lower TEER and cilia beat frequency (CBF) compared to BEGM media. PneumaCultTM media cultures exhibited significantly increased FOXJ1 expression, more ciliated cells with a larger active area, increased intracellular mucins, and increased calcium-activated chloride channel current. However, there were no significant changes in viral RNA or host antiviral responses. Conclusion: There are distinct structural and functional differences in CRpBECs cultured in the two commonly used ALI differentiation media. Such factors need to be taken into consideration when designing and comparing CRpBECs ALI experiments. 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/2023.02.17.529005v1?rss=1 Authors: He, H., Yang, J., He, Y., Li, Z., Fu, C., Zhang, D., Li, M., Lu, A., Dong, J., Liu, J., Gu, H., Shen, S. Abstract: Ulva prolifera is the dominant species of "green tide", and has higher tolerance to environmental stresses such as temperature. However, the molecular mechanisms are still unclear. Here, transcriptome analysis, Western blot and RT-qPCR analysis of U. prolifera suggested that, under temperature stresses (4{degrees}C, 36{degrees}C), the expression of PCNA and CyclinA was promoted, and the MAPK signaling was activated. Besides, the results showed that PCNA interacted with CyclinA. Interestingly, the expression of miR-2916, which was predicted to bind PCNA at -552~-772, was negatively correlated with the expression of PCNA under temperature stresses (4{degrees}C, 36{degrees}C). In addition, the results showed that low temperature (4{degrees}C) had no obvious effect on the survival, the formation of cell walls, and the division of protoplasts. However, high temperature (36{degrees}C) had obvious effect on them. PCNA inhibitors increased the sensitivity of the protoplasts under temperature stresses. Together, our results suggested PCNA regulating the proliferation in response to the temperature stress of U. prolifera was associated with miR-2916/PCNA/CyclinA/MAPK pathway. In conclusion, the study preliminarily illuminates the molecular mechanism in response to temperature stress of U. prolifera, and may provide a new insight for prevention of green tide. 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/2023.02.16.525994v1?rss=1 Authors: Luo, Y., Li, J., Xiong, Y. Abstract: Purpose: Glioblastoma (GBM) is the most aggressive and common form of brain cancer in adults. GBM is characterised by poor survival as the lack of effective therapies. This research aims to detect the roles of SNAREs in GBM and improve our knowledge of targeting therapy for GBM. Materials and methods: the expression of SNAREs and their correlation with overall survival (OS) in GBM are investigated using the GEPIA. The level of BET1 in GBM cell lines was tested by RT-qPCR, and its biological functions in GBM cells were tested by Transwell assay and CCK8 kit. The effect of BET1 on the location of MMP14 is identified by Immunofluorescence. Results: The expression profile of SNARE family members in GBM tissue is changed dramatically. Among them, the mRNA levels of BET1 and VAMP3 are up-regulated, and their expression negatively correlates with OS. BET1 is also increased in GBM Cell Lines, and it is required for efficient GBM cell migration and invasion partly because it mediates the transport of MMP14 to the plasma membrane. Conclusion: GBM has highly diffusive and infiltrative ability in nature, making complete surgical resection almost impossible. Our data shows that BET1 is highly expressed in GBM tissue, negatively correlated with OS, and essential for GBM cell migration and invasion. These results indicate that SNARE BET1 may present a potential target for GBM treatment. 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/2023.02.15.528337v1?rss=1 Authors: Toro, C. A., Johnson, K. E., Hansen, J., Siddiq, M. M., Vasquez, W., Zhao, W., Graham, Z. A., Saez, J. C., Iyengar, R., Cardozo, C. P. Abstract: Membrane channels such as connexins (Cx), pannexins (Panx) and P2X7 receptors (P2X7R) are permeable to calcium ions and other small molecules such as ATP and glutamate. Release of ATP and glutamate through these channels is a key mechanism driving tissue response to traumas such as spinal cord injury (SCI). Boldine, an alkaloid isolated from the Chilean boldo tree, blocks both Cx hemichannels (HC) and Panx. To test if boldine could improve function after SCI, boldine or vehicle was administered to treat mice with a moderate severity contusion-induced SCI. Boldine led to greater spared white matter and increased locomotor function as determined by the Basso Mouse Scale and horizontal ladder rung walk tests. Boldine treatment reduced immunostaining for markers of activated microglia (Iba1) and astrocytic (GFAP) markers while increasing that for axon growth and neuroplasticity (GAP-43). Cell culture studies demonstrated that boldine blocked glial HC, specifically Cx26 and Cx30, in cultured astrocytes and blocked calcium entry through activated P2X7R. RT-qPCR studies showed that boldine treatment reduced expression of the chemokine Ccl2, cytokine IL-6 and microglial gene CD68, while increasing expression of the neurotransmission genes Snap25 and Grin2b, and Gap-43. Bulk RNA sequencing (of the spinal cord revealed that boldine modulated a large number of genes involved in neurotransmission in in spinal cord tissue just below the lesion epicenter at 14 days after SCI. Numbers of genes regulated by boldine was much lower at 28 days after injury. These results indicate that boldine treatment ameliorates injury and spares tissue to increase locomotor function. 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/2023.02.08.527757v1?rss=1 Authors: Ridley, R. B., Bowman, B. M., Lee, J., Walsh, E., Massengill, M. T., Lewin, A. S., Ildefonso, C. J. Abstract: The advanced form of AMD, geographic atrophy, is associated with increased RPE oxidative stress and chronic inflammation. Here we evaluated the effects of delivering an anti-inflammatory viral gene by an AAV-vector in a mouse model of geographic atrophy. We measured changes in retinal function, structure, and morphology over nine months with electroretinography, optical coherence tomography, and fundoscopy, respectively. In addition, we used retinal tissue to quantify changes in markers of inflammation by multiplex ELISA, RT-qPCR, and immunofluorescence staining. Our AAV significantly delayed the loss of retinal function and structure and decreased retinal inflammation compared to the control AAV treatment. Our results suggest that modulating retinal inflammation could significantly slow the progression of geographic atrophy. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

A new research paper was published on the cover of Aging (Aging-US) Volume 14, Issue 22, entitled, “Glutaminase inhibitors rejuvenate human skin via clearance of senescent cells: a study using a mouse/human chimeric model.” Skin aging caused by various endogenous and exogenous factors results in structural and functional changes to skin components. However, the role of senescent cells in skin aging has not been clarified. In this new study, researchers Kento Takaya, Tatsuyuki Ishii, Toru Asou, and Kazuo Kishi, from the Department of Plastic and Reconstructive Surgery at the Keio University School of Medicine, evaluated the effects of the glutaminase inhibitor BPTES (bis-2-(5-phenylacetamido-1, 3, 4-thiadiazol-2-yl)ethyl sulfide) on human senescent dermal fibroblasts and aged human skin to elucidate the function of senescent cells in skin aging. “[...] we utilized plastic surgery to create an experimental mouse/human chimeric model in which intraoperatively obtained human whole skin layers were transplanted into nude mice using previously described methods [25] and evaluated the anti-aging effects of BPTES on real human skin.” Primary human dermal fibroblasts (HDFs) were induced to senescence by long-term passaging, ionizing radiation, and treatment with doxorubicin, an anticancer drug. Cell viability of HDFs was assessed after BPTES treatment. A mouse/human chimeric model was created by subcutaneously transplanting whole skin grafts from aged humans into nude mice. The model was treated intraperitoneally with BPTES or vehicle for 30 days. Skin samples were collected and subjected to reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blotting, and histological analysis. BPTES selectively eliminated senescent dermal fibroblasts regardless of the method used to induce senescence; aged human skin grafts treated with BPTES exhibited increased collagen density, increased cell proliferation in the dermis, and decreased aging-related secretory phenotypes, such as matrix metalloprotease and interleukin. These effects were maintained in the grafts 1 month after termination of the treatment. In conclusion, selective removal of senescent dermal fibroblasts can improve the skin aging phenotype, indicating that BPTES may be an effective novel therapeutic agent for skin aging. “In summary, our results indicate that selective clearance of aging dermal fibroblasts by BPTES ameliorates skin senescence-related changes and that aging dermal fibroblasts may play an important role in the skin aging process. Therefore, senescent cell eliminators for aging skin cells may be an effective option for treating skin aging.” DOI: https://doi.org/10.18632/aging.204391 Corresponding Author: Kento Takaya - kento-takaya312@keio.jp Sign up for free Altmetric alerts about this article: https://aging.altmetric.com/details/email_updates?id=10.18632%2Faging.204391 About Aging-US Launched in 2009, Aging-US publishes papers of general interest and biological significance in all fields of aging research and age-related diseases, including cancer—and now, with a special focus on COVID-19 vulnerability as an age-dependent syndrome. Topics in Aging-US go beyond traditional gerontology, including, but not limited to, cellular and molecular biology, human age-related diseases, pathology in model organisms, signal transduction pathways (e.g., p53, sirtuins, and PI-3K/AKT/mTOR, among others), and approaches to modulating these signaling pathways. Please visit our website at https://www.Aging-US.com​​ and connect with us: SoundCloud - https://soundcloud.com/Aging-Us Facebook - https://www.facebook.com/AgingUS/ Twitter - https://twitter.com/AgingJrnl Instagram - https://www.instagram.com/agingjrnl/ YouTube - https://www.youtube.com/agingus​ LinkedIn - https://www.linkedin.com/company/aging/ Pinterest - https://www.pinterest.com/AgingUS/ For media inquiries, please contact media@impactjournals.com

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.24.517673v1?rss=1 Authors: Stylianakis, E., Festenstein, R., Vannier, J.-B. Abstract: TElomeric Repeat-containing RNA are long non-coding RNAs generated from the telomeres. TERRAs are essential for the establishment of heterochromatin marks at telomeres, which serve for the binding of Heterochromatin Protein 1 (HP1), a protein family of epigenetic modifiers involved with chromatin compaction and gene silencing. While HP1{gamma} is enriched on gene bodies of actively transcribed human and mouse genes, it is unclear if its transcriptional role is important for HP1{gamma} function in telomere cohesion and telomere maintenance. We aimed to study the effect of mouse HP1{gamma} on the transcription of telomere factors and molecules that can affect telomere maintenance. We investigated the telomere function of HP1{gamma} by using deficient mouse embryonic fibroblasts (MEFs) deriving from 13.5 embryonic day embryos compared to their litter mate controls. We used gene expression analysis of HP1{gamma} deficient MEFs and validated the molecular and mechanistic consequences of HP1{gamma} loss by telomere FISH, immunofluorescence, RT-qPCR and DNA-RNA Immunoprecipitation (DRIP). Loss of HP1{gamma} in primary MEFs leads to a downregulation of various telomere and telomere-accessory transcripts, including shelterin protein TRF1. Its downregulation is associated with increased telomere replication stress and DNA damage ({gamma}H2AX), effects more profound in females. We suggest that the source for the impaired telomere maintenance is a consequence of increased telomeric DNA-RNA hybrids and TERRAs arising at and from mouse chromosomes 18 and X. Our results suggest an important transcriptional control by mouse HP1{gamma} of various telomere factors including TRF1 protein and TERRAs that has profound consequences on telomere stability, with a potential sexually dimorphic nature. 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/2022.11.09.515885v1?rss=1 Authors: Biber, J. C., Sullivan, A., Brazzo, J. A., Krajnik, A., Heo, Y., Poppenberg, K. E., Tutino, V. M., Heo, S.-J., Kolega, J., Lee, K., Bae, Y. Abstract: Stiffened arteries are a pathology of atherosclerosis, hypertension, and coronary artery disease and a key risk factor for cardiovascular disease events. The increased stiffness of arteries triggers the hypermigration and hyperproliferation of vascular smooth muscle cells (VSMCs), leading to neointimal hyperplasia and accelerated neointima formation, but the mechanism of this trigger is not known. Our analyses of whole-transcriptome microarray data sets from mouse VSMCs cultured on stiff hydrogels simulating arterial pathology and from injured mouse femoral arteries revealed 80 genes that were differentially regulated (74 upregulated and 6 downregulated) relative to expression in control VSMCs cultured on soft hydrogels and in uninjured femoral arteries. A functional enrichment analysis revealed that these stiffness-sensitive genes are linked to cell cycle progression and proliferation. Furthermore, we found that survivin, a member of the inhibitor of apoptosis protein family, mediates stiffness-sensitive cell cycling and proliferation in vivo and in vitro as determined by gene network and pathway analyses, RT-qPCR, and immunoblotting. The stiffness signal is mechanotransduced via FAK and Rac signaling to regulate survivin expression, establishing a regulatory pathway for how the stiffness of the cellular microenvironment affects VSMC behaviors. Our findings indicate that survivin is necessary for VSMC cycling and proliferation and regulates stiffness-responsive phenotypes. 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/2022.11.04.515175v1?rss=1 Authors: Obis, E., Sol, J., Andres-Benito, P., Martin-Gari, M., Mota-Martorell, N., Galo-Licona, J. D., Pinol-Ripoll, G., Portero-Otin, M., Ferrer, I., Jove, M., Pamplona, R. Abstract: Aims: Non-targeted lipidomics analysis was conducted in post-mortem human frontal cortex area 8 (GM) and white matter of the frontal lobe centrum semi-ovale (WM) to identify lipidomes in middle-aged individuals with no neurofibrillary tangles and senile plaques, and cases at progressive stages of sporadic Alzheimer's disease (sAD). Methods: Lipidomic analysis using an LC-MS/MS platform was carried out in selected cases with suitable post-mortem lacking co-morbidities and concomitant brain pathologies. Complementary data were obtained using RT-qPCR and immunohistochemistry. Results: The WM shows an adaptive lipid phenotype resistant to lipid peroxidation, characterized by a lower fatty acid unsaturation, peroxidizability index, and higher ether lipid content than the GM. Changes in the lipidomic profile more marked in the WM than in GM occur in AD with disease progression. WM alterations are characterized by a decline in the content of branched fatty acid esters of hydroxy fatty acids (FAHFA), diacylglycerols (DG), triacylglycerols (TG), glycerophospholipids (GP) (especially ether lipids), and sphingolipids (especially sulfatides, ceramides, and glycosphingolipids). The GM acquires a fatty acid profile prone to peroxidation in sAD, while WM reinforces its peroxidation-resistant trait. Transcriptomic data point to additional defects of peroxisomal function. Conclusions: Four functional categories are associated with the different lipid classes affected in sAD: membrane structural composition, bioenergetics, antioxidant protection, and bioactive lipids, with deleterious consequences affecting both neurons and glial cells favoring disease progression. 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/2022.11.02.514828v1?rss=1 Authors: Fourneaux, C., Racine, L., Koering, C., Dussurgey, S., Vallin, E., Moussy, A., Parmentier, R., Brunard, F., Stockholm, D., Modolo, L., Picard, F., Gandrillon, O., Paldi, A., Gonin-Giraud, S. Abstract: Cell differentiation requires the integration of two opposite processes, a stabilizing cellular memory, especially at the transcriptional scale, and a burst of gene expression variability which follows the differentiation induction. Therefore, the actual capacity of a cell to undergo phenotypic change during a differentiation process relies upon a modification in this balance which favors change-inducing gene expression variability. However, there are no experimental data providing insight on how fast the transcriptomes of identical cells would diverge on the scale of the very first two cell divisions during the differentiation process. In order to quantitatively address this question, we developed different experimental methods to recover the transcriptomes of related cells, after one and two divisions, while preserving the information about their lineage at the scale of a single cell division. We analyzed the transcriptomes of related cells from two differentiation biological systems (human CD34+ cells and T2EC chicken primary erythrocytic progenitors) using two different single-cell transcriptomics technologies (sc-RT-qPCR and scRNA-seq). We identified that the gene transcription profiles of differentiating sister-cells are more similar to each-other than to those of non related cells of the same type, sharing the same environment and undergoing similar biological processes. More importantly, we observed greater discrepancies between differentiating sister-cells than between self-renewing sister-cells. Furthermore, a continuous increase in this divergence from first generation to second generation was observed when comparing differentiating cousin-cells to self renewing cousin-cells. Our results are in favor of a continuous and gradual erasure of transcriptional memory during the differentiation process. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

În cadrul ediției de pe 10 mai 2022 a emisiunii #știința360 de pe Radio România Cultural, Dr. Marius Geantă, Președintele Centrului pentru Inovație în Medicină #inomed, a comentat topul săptămânal Esențial Covid-19 de pe Raportuldegardă.ro. Conform unui studiu bazat pe monitorizarea apelor uzate, deși Omicron a devenit rapid varianta dominantă, aceasta nu a determinat dispariția variantei Delta – care a continuat să fie depistată fără modificări majore. Deoarece varianta Omicron a prezentat caracteristici epidemiologice diferite față de variantele precedente, o echipă de cercetători din Israel a dezvoltat în scurt timp un test RT-qPCR specific pentru detectarea variantei Omicron, utilizându-l pe probe din ape uzate. Deși așteptarea era ca detectarea variantei Delta să se diminueze pe măsură ce Omicron devine tot mai prevalentă, rezultatele studiului demonstrează continuarea circulației variantei Delta. Cercetătorii au dezvoltat un model de evaluare a dinamicii celor două variante pe baza datelor obținute inițial care sugerează că semnalele de detectare ale variantei Omicron vor continua să scadă până la eliminarea completă, în timp ce varianta Delta își va menține nivelul de detecție actual. Dacă acest lucru se va întâmpla, tiparul atipic de circulație a variantei Delta ar putea determina reapariția unui val Delta sau emergența unei noi variante îngrijorătoare. Mai multe detalii - https://bit.ly/38uggqb

Conclusiones 1- El rendimiento analítico de RT-qPCR podría controlarse utilizando metodologías adecuadas, otros componentes de las pruebas moleculares son mucho más difíciles de evaluar y estandarizar, como la adecuación y reproducibilidad de la recolección de muestras (fuente de la muestra y técnica de recolección), el impacto de la muestra almacenamiento, transporte y pretratamiento, y variaciones en el proceso de detección molecular, como la agrupación de muestras. Con RDT. 2- Al establecer estándares de desempeño es primordial para garantizar la estandarización y armonización, debemos considerar, según la experiencia acumulada durante la respuesta a la pandemia COVID-19, que los requisitos de prueba podrían diferir entre entornos y aplicaciones. Por ejemplo, una prueba utilizada para clasificar a los pacientes con sospecha de COVID-19 en el hospital puede requerir un estándar de rendimiento diferente en comparación con una prueba de 'activación' utilizada para descartar la infección por SARS-CoV-2 en personas asintomáticas que asisten a eventos públicos. De manera similar, las pruebas serológicas utilizadas en el punto de atención con fines de vigilancia pueden requerir diferentes estándares de desempeño en comparación con la serología formal realizada durante un estudio clínico. 3- Los dos primeros estudios no solo brindan información valiosa con respecto al desempeño real de un gran número de kits de PDR comerciales ampliamente distribuidos, sino que también reportan un marco de evaluación factible que podría adoptarse ampliamente. 4- La sorprendente variabilidad en el rendimiento de diferentes ensayos comerciales destaca la importancia del control de calidad y la garantía de calidad en una situación de pandemia. El rápido despliegue y adopción de pruebas de laboratorio resultó en desafíos notables, como la falta de materiales de referencia certificados, incertidumbres de medición, falta de correlatos epidemiológicos o clínicos y falta de estandarización y armonización, por nombrar algunos. 5- De cara al futuro, las comunidades científicas y de salud pública deben desarrollar más soluciones para cerrar las brechas identificadas durante la respuesta mundial del laboratorio a la pandemia de COVID-19. Esto requiere un enfoque holístico que reúna a una amplia gama de partes interesadas de los campos médico, de laboratorio, de salud pública, industrial y regulatorio. 6- Existe una necesidad urgente de un marco aceptado a nivel mundial que informará el desarrollo, la validación y la implementación de nuevos ensayos frente a las amenazas emergentes para la salud pública. 7- Tener soluciones adecuadas para garantizar el rendimiento de las pruebas debe ser una parte integral de la preparación para emergencias nacional y mundial para garantizar una respuesta de laboratorio rápida y sólida a futuras pandemias. REFERENCIA. https://www.eurosurveillance.org/docserver/fulltext/eurosurveillance/26/45/eurosurv-26-45-1.pdf?expires=1637286854&id=id&accname=guest&checksum=C36D79C93FA4C9DB5D31350BB9045906 https://www.eurosurveillance.org/content/10.2807/1560-7917.ES.2021.26.45.2101050 https://journals.lww.com/co-pulmonarymedicine/Fulltext/2021/05000/COVID_19_new_diagnostics_development__novel.4.aspx

Greater adherence to Mediterranean diet associated with better cognitive performance in older individuals University of South Australia, July 13, 2021 According to news originating from Adelaide, Australia, by NewsRx correspondents, research stated, “Adherence to a Mediterranean diet is associated with higher cognitive function and reduced risk of dementia in Mediterranean populations. However, few studies have investigated the association between Mediterranean diet adherence and cognition in populations outside of the Mediterranean basin.” Our news journalists obtained a quote from the research from the University of South Australia, “Furthermore, it is currently unknown whether the association between Mediterranean diet adherence and cognitive function differs between middle-aged and older individuals. Cross-sectional (n = 894) and longitudinal (n = 530) multivariable analyses were undertaken using data from community-dwelling adults from the Maine-Syracuse Longitudinal Study (MSLS). Mediterranean diet adherence was measured by applying a literature-based Mediterranean diet score to food frequency questionnaire data. Cognitive function was assessed with a battery of tests and composites scores were computed for global cognitive function, Visual-Spatial Organisation and Memory, verbal memory, working memory, scanning and tracking and abstract reasoning. No cross-sectional associations between Mediterranean diet adherence and cognitive function were detected. Over a period of five years, higher adherence to a Mediterranean diet was associated with improvements in Global Cognitive Function, Visual-Spatial Organisation and Memory and scanning and tracking in participants >= 70 years.” According to the news editors, the research concluded: “No significant longitudinal associations were observed for participants Conclusion: Our findings suggest that higher adherence to a Mediterranean diet is associated with better cognitive performance, and therefore less cognitive decline, in older but not middle-aged individuals.” This research has been peer-reviewed.     Coffee and veggies may protect against COVID-19 Northwestern University, July 20, 2021 Sip a venti dark roast and eat a salad. A new Northwestern Medicine study shows coffee consumption and eating lots of vegetables may offer some protection against COVID-19. The authors believe this is the first study using population data to examine the role of specific dietary intake in prevention of COVID-19. "A person's nutrition impacts immunity," said senior author Marilyn Cornelis, associate professor of preventive medicine at Northwestern University Feinberg School of Medicine. "And the immune system plays a key role in an individual's susceptibility and response to infectious diseases, including COVID-19." Being breastfed may also offer protection as well as eating less processed meats, the study found. "Besides following guidelines currently in place to slow the spread of the virus, we provide support for other relatively simple ways in which individuals can reduce their risk and that is through diet and nutrition," Cornelis said.  The paper on nutrition and COVID-19 protection was published recently in the journal Nutrients. One or more cups of coffee per day was associated with about a 10% decrease in risk of COVID-19 compared to less than one cup per day. Consumption of at least 0.67 servings per day of vegetables (cooked or raw, excluding potatoes) was associated with a lower risk of COVID-19 infection. Processed meat consumption of as little as 0.43 servings per day was associated with a higher risk of COVID-19. Having been breastfed as a baby reduced the risk 10% compared to not having been breastfed.  While the study shows diet appears to modestly reduce disease risk, the Centers for Disease Control and Prevention recommends vaccines as the most effective way to prevent COVID-19 disease, especially severe illness and death. COVID-19 vaccines also reduce the risk of people spreading the virus that causes COVID-19. Thus far, most COVID-19 research has focused on individual factors assessed after a positive COVID-19 test. Individuals with suppressed immune systems such as the elderly and those with existing comorbidities including cardiovascular diseases, hypertension, diabetes and obesity, are more likely to experience severe outcomes of COVID-19.  But other than weight management, less attention has focused on other modifiable risk factors preceding COVID-19 infection, said Cornelis, who studies how diet and nutrition contribute to chronic disease.  Dr. Thanh-Huyen Vu, the study's first author and a research associate professor of medicine at Northwestern, is now leading analyses to determine whether these protective diet behaviors are specific to COVID or respiratory infections more broadly.  Exact mechanisms linking these diet factors to COVID are unknown.  "Coffee is a major source of caffeine, but there are also dozens of other compounds that may potentially underlie the protective associations we observed," Cornelius said. "Associations with processed meat, but not red meat, point to non-meat factors." Using data from the UK Biobank, researchers examined the associations between dietary behaviors measured in 2006-2010 and COVID-19 infections in March to December 2020, before vaccines were available. They focused on 1) diet factors for which data were available and previously implicated in immunity based on human and animal studies; 2) self-reported intakes of coffee, tea, vegetables, fruit, fatty fish, processed meat and red meat. An early-life exposure to breastmilk also was analyzed.  Among the 37,988 participants tested for COVID-19 and included in the study, 17% tested positive.  The observational nature of the UK Biobank research limits the extent to which mechanisms of protection can be tested, Cornelis said. However, much of her nutrition research uses genetics, and with all UK Biobank participants currently genotyped, she hopes to use this information to gain better insight into how diet and nutrition offer protection from the disease.   Biologic age reversed with lifestyle improvement plus supplements Institute for Functional Medicine (Seattle), July 14 2021.  The April 15, 2021 issue of Aging published the results of an eight-week randomized trial which resulted in a reduction in biologic age among men who participated in lifestyle changes and consumed nutritional supplements. "The combined intervention program was designed to target a specific biological mechanism called DNA methylation, and in particular the DNA methylation patterns that have been identified as highly predictive of biological age,” explained lead author Kara Fitzgerald, ND. “We suspect that this focus was the reason for its remarkable impact.”  The trial included 38 men between the ages of 50 and 72 years. Eighteen participants consumed a plant-based, low carbohydrate diet that included limited nutrient-dense animal proteins. The diet was supplemented with a vegetable and fruit powder and the probiotic Lactobacillus plantarum 299v. The group was advised to participate in a minimum of 30 minutes of exercise daily and to perform breathing exercises twice per day for stress reduction.  According to the Horvath DNAmAge clock, which evaluates DNA methylation patterns as a marker of biologic age, men who participated in the lifestyle program had scores that averaged 1.96 younger at the end of the program in comparison with the beginning, while control participants scored 1.27 years older. Additionally, triglycerides were reduced in the lifestyle program group. “To our knowledge, this is the first randomized controlled study to suggest that specific diet and lifestyle interventions may reverse Horvath DNAmAge epigenetic aging in healthy adult males,” the authors announced.  “These early results appear to be consistent with, and greatly extend, the very few existing studies that have so far examined the potential for biological age reversal,” Dr Fitzgerald commented. “And it is unique in its use of a safe, non-pharmaceutical dietary and lifestyle program, control group, and the extent of the age reduction."       Research suggests L-carnitine could aid burn recovery Anhui Medical University (China), July 12, 2021 According to news reporting from First Affiliated Hospital of Anhui Medical University research stated, “Impaired hepatic fatty acid metabolism and persistent mitochondrial dysfunction are phenomena commonly associated with liver failure. Decreased serum levels of L-carnitine, a amino acid derivative involved in fatty-acid and energy metabolism, have been reported in severe burn patients.” Our news editors obtained a quote from the research from First Affiliated Hospital of Anhui Medical University: “The current study aimed to evaluate the effects of L-carnitine supplementation on mitochondrial damage and other hepatocyte injuries following severe burns and the related mechanisms. Serum carnitine and other indicators of hepatocytic injury, including AST, ALT, LDH, TG, and OCT, were analyzed in severe burn patients and healthy controls. A burn model was established on the back skin of rats; thereafter, carnitine was administered, and serum levels of the above indicators were evaluated along with Oil Red O and TUNEL staining, transmission electron microscopy, and assessment of mitochondrial membrane potential and carnitine palmitoyltransferase 1 (CPT1) activity and expression levels in the liver. HepG2 cells pretreated with the CPT1 inhibitor etomoxir were treated with or without carnitine for 24 h. Next, the above indicators were examined, and apoptotic cells were analyzed via flow cytometry. High-throughput sequencing of rat liver tissues identified several differentially expressed genes (Fabp4, Acacb, Acsm5, and Pnpla3) were confirmed using RT-qPCR. Substantially decreased serum levels of carnitine and increased levels of AST, ALT, LDH, and OCT were detected in severe burn patients and the burn model rats. Accumulation of TG, evident mitochondrial shrinkage, altered mitochondrial membrane potential, decreased ketogenesis, and reduced CPT1 activity were detected in the liver tissue of the burned rats. Carnitine administration recovered CPT1 activity and improved all indicators related to cellular and fatty acid metabolism and mitochondrial injury. Inhibition of CPT1 activity with etomoxir induced hepatocyte injuries similar to those in burn patients and burned rats; carnitine supplementation restored CPT1 activity and ameliorated these injuries. The expression levels of the differentially expressed genes Fabp4, Acacb, Acsm5, and Pnpla3 in the liver tissue from burned rats and etomoxir-treated hepatocytes were also restored by treatment with exogenous carnitine.” According to the news editors, the research concluded: “Exogenous carnitine exerts protective effects against severe burn-induced cellular, fatty-acid metabolism, and mitochondrial dysfunction of hepatocytes by restoring CPT1 activity.”     Red blood cell ‘traffic' contributes to changes in brain oxygenation   Penn State University, July 19, 2021 Adequate blood flow supplies the brain with oxygen and nutrients, but the oxygenation tends to fluctuate in a distinct, consistent manner. The root of this varied activity, though, is poorly understood. Now, Penn State researchers have identified one cause of the fluctuations: inherent randomness in the flow rate of red blood cells through tiny blood vessels called capillaries. According to the researchers, this randomness could have potential implications for understanding the biological build-up mechanisms underlying neurodegenerative diseases, such as Alzheimer's disease. They published their findings in PLOS Biology today. “These oxygenation fluctuations also occur in other tissues, like muscle,” said Patrick Drew, Huck Distinguished Associate Professor of Engineering Science and Mechanics, Neurosurgery and Biomedical Engineering. “The question we had was: Are these fluctuations caused by neural activity or something else?” The fluctuations resemble 1/f-like noise, a statistical pattern showing large fluctuations made up of many small fluctuations and naturally occurring in a variety of phenomena, from stock-market prices to river heights. The researchers investigated the fluctuations in mice due to their brains' similarities to those of humans, according to Drew, who also serves as associate director of the Penn State Neuroscience Institute. First, the researchers monitored the blood flow, oxygenation and electrical signals produced by brain activity—the first time the latter two had been tracked simultaneously, according to Drew—in awake mice. They collected the data as mice moved on a spherical treadmill for up to 40 minutes at a time. Next, to investigate the relationship between brain activity and oxygenation fluctuations, the researchers used pharmacological compounds to temporarily and reversibly silence neural signals in the mice's brains. Despite the silencing, the fluctuations continued, showing little correlation between neural activity and oxygenation. The passage of red blood cells, however, told a different story. Using two-photon laser scanning microscopy, an imaging technique used to visualize cells deep inside living tissue, the researchers could visualize the passage of individual red blood cells through capillaries. “It's like traffic,” Drew said. “Sometimes there are a lot of cars going by, and the traffic gets plugged up, and sometimes there aren't. And red blood cells go either way when they approach a junction, so this random flow can lead to bottlenecks and stalls in the vessel.” Importing experimental data into a statistical model allowed the researchers to run further simulations and make inferences based on massive amounts of data produced by the model. The researchers discovered that these random red blood cell stoppages contributed to the fluctuations in oxygenation, further supporting a relationship between the flow of red blood cells through capillaries and the tiny changes in oxygenation that formed larger trends. Better understanding the regulation of blood flow and subsequent transport of oxygen can help researchers improve medical technology and explore causes of diseases such as Alzheimer's, according to Drew. While the researchers identified the link between red blood cell transport and oxygenation, further research is needed to investigate additional contributors to oxygenation fluctuations that could play a role in neurodegenerative diseases. Kyle Gheres, a graduate student in the intercollege Graduate Program in Molecular Cellular and Integrative Biosciences, also contributed to this paper. Qingguang Zhang, assistant research professor of engineering science and mechanics, served as first author on the paper. This work was supported by the National Institutes of Health.     EGCG in Green Tea inhibits the growth of breast cancer cells Chonbuk National University School of Medicine (S Korea), July 21, 2021   Findings on Breast Cancer Reported by Researchers at Chonbuk National University School of Medicine(Epigallocatechin gallate inhibits the growth of MDA-MB-231 breast cancer cells via inactivation of the b-catenin signaling pathway) According to news reporting originating in Chonbuk, South Korea, research stated, "Epigallocatechin gallate (EGCG), a major constituent of green tea, has potential as a treatment for a variety of diseases, including cancer. EGCG induces apoptosis and inhibits tumorigenesis through multiple signaling pathways in breast cancer cells. b-catenin signaling modulators could be useful in the prevention and therapy of breast cancer." The news reporters obtained a quote from the research from the Chonbuk National University School of Medicine, "However, the precise anticancer effect of EGCG through the b-catenin signaling pathway in breast cancer is unclear. The present study investigated the association between b-catenin expression and clinicopathological factors of breast cancer patients, and the effect of EGCG on b-catenin expression in breast cancer cells. b-catenin expression was analyzed according to the clinicopathological factors of 74 patients with breast cancer. All patients were females diagnosed with invasive ductal carcinoma. Western blot analysis revealed that b-catenin was expressed at higher levels in breast cancer tissue than in normal tissue. b-catenin expression was associated with lymph node metastasis (p=0.04), tumor-node-metastasis stage (p=0.03) and estrogen receptor status (p

Listen to the latest oncology-focused research published in this week’s issue of Oncotarget, Volume 12, Issue 10. View the complete issue: https://www.oncotarget.com/archive/v12/i10/ COVER PAPER: “Inhibitory effects of Tomivosertib in acute myeloid leukemia” https://doi.org/10.18632/oncotarget.27952 EDITORIAL: “Raman spectroscopy determination of the mineral characteristics of microcalcifications in breast cancer, a way towards an improved screening approach” https://doi.org/10.18632/oncotarget.27912 (PDF Download) EDITORIAL: “Reactive oxygen species in leukemias: maintaining cancer cell proliferation via redox signaling and changing metabolic homeostasis” https://doi.org/10.18632/oncotarget.27913 (PDF Download) RESEARCH PAPER: “Genome wide DNA methylation landscape reveals glioblastoma’s influence on epigenetic changes in tumor infiltrating CD4+ T cells” https://doi.org/10.18632/oncotarget.27955 RESEARCH PAPER: “Association between miRNA signatures in serum samples from epidermal growth factor inhibitor treated patients and skin toxicity” https://doi.org/10.18632/oncotarget.27953 RESEARCH PAPER: “Perioperative changes in the plasma metabolome of patients receiving general anesthesia for pancreatic cancer surgery” https://doi.org/10.18632/oncotarget.27956 RESEARCH PAPER: “Transcriptome analyses of urine RNA reveal tumor markers for human bladder cancer: validated amplicons for RT-qPCR-based detection” https://doi.org/10.18632/oncotarget.27954 Keywords - acute myeloid leukemia, breast cancer, bladder cancer, leukemia, DNA methylation, tumor, glioblastoma, miRNA, skin toxicity, pancreatic cancer, cancer, science, research, oncology About Oncotarget Oncotarget is a bi-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: SoundCloud - https://soundcloud.com/oncotarget Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget LinkedIn - https://www.linkedin.com/company/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/oncotargetyoutube Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget Oncotarget is published by Impact Journals, LLC. Please visit https://www.impactjournals.com/ or connect with @ImpactJrnls Media Contact MEDIA@IMPACTJOURNALS.COM 18009220957

Meritve SARS-CoV-2 v čistilnih napravah so poleg testov ključne za razumevanje njegove razširjenosti. Oddelek za biotehnologijo in sistemsko biologijo NIB je že marca lani pilotno začel meriti navzočnost novega virusa v odpadnih vodah. V okviru mednarodne medlaboratorijske primerjave 21 državnih meroslovnih inštitutov in laboratorijev iz 16 držav so dokazali, da je natančno merjenje količine virusne RNA SARS-CoV-2 moč doseči z kvantitativne PCR s povratno transkripcijo oz. RT-qPCR. Od lani pri nas to počnejo na osmih čistilnih napravah - Ljubljana, Domžale-Kamnik, Kranj, Koper, Rogaška Slatina, Velenje-Šoštanj, Celje in Maribor. Temu pravijo, kot v pogovoru pojasni dr. Denis Kutnjak, Nacionalni inštitut za biologijo, epidemiologija odpadne vode. Za bolj nemoten potek tovrstne v spopadu z epidemijo novega virusa pomembno metodo, bo mogoče pomagalo marca sprejeto priporočilo Komisije EU o monitoringu COVID-19 in njegovih različic v odpadnih vodah EU. Vsaj pri nas je NIB doslej to delo v epidemiološkem nadzoru opravljal v okviru že obstoječega raziskovalnega programa, in razen posebnih sredstev ARRS, od države za to ni prejel še nobenih finančnih nadomestil. Na fotografiji postopek koncentriranja vzorcev odpadne vode v laboratoriju pred analizo z metodo verižne reakcije s polimerazo z obratnim prepisovanjem. Vir: Aleš Rosa, NIB

Switching New Zealand's required method of Covid-19 testing could be on the horizon as the roll-out of voluntary saliva tests widens.Auckland Airport workers are now being offered saliva testing in a bid to add another layer of protection.It comes after the Government last month rolled out the additional testing method for border workers in quarantine facilities.While the Ministry of Health say at this stage there is still no indication that saliva PCR testing could replace the mandatory nasopharyngeal testing, health experts say it's only a matter of time.Otago University epidemiologist Professor Michael Baker said once everyone was convinced the performance of saliva testing was adequate it could certainly replace nasal swab testing."It means we could contemplate the idea of having daily tests as a way of increasing the speed of outbreak protection," Baker said.Baker said it was good New Zealand had multiple laboratories testing out these technologies as the benefits were huge."Saliva testing is a lot less invasive for people ... potentially switching to saliva testing, especially for workforce who are already feeling more vulnerable, could make the job that much easier."Mary-Liz Tuck, Auckland Airport general manager of corporate services, said it made asymptomatic testing simple and comfortable for airport and border workers, while providing the highest standard of protection for the community."We want to see as many protective layers as possible for our people and our community," Tuck said.The airport teamed up with New Zealand business Rako Science that established the Covid-19 surveillance testing in New Zealand using the Shield saliva test developed at the University of Illinois.A collection site in Auckland Airport's international terminal had now been set up to provide the saliva testing and was set to run for three months while scientists measured the effectiveness.Airport staff taking part in the saliva tests were doing so on a voluntary basis, along with the mandatory nasal-swab testing required by the Government's border policies.A Ministry of Health spokeswoman said PCR nasopharyngeal swabbing method was still considered the gold standard for Covid testing as it detects the virus the most effectively."This swab type will obtain the optimal specimen and is the preferred collection method for both symptomatic and asymptomatic testing due to its higher sensitivity in detecting the virus."She said voluntary saliva testing also recommenced this week at the Auckland quarantine facility and at the managed isolation and quarantine facility in Christchurch.Rako Science is using the RT-qPCR saliva-testing protocol developed by University of Illinois which has conducted 1.3 million on-campus tests.Rako Science's Chief Science officer, Dr Stephen Grice, said Rako's science committee had briefed the Ministry of Health in December last year that it had successfully validated and accredited the saliva test for New Zealand.He said they recently provided the ministry with additional data which confirmed the test was as accurate as tests using nasal swabs."Rako Science has the processing capacity for 10,000 tests per day and the sample collection system does not depend on medical professionals to administer the test."Saliva testing for Covid-19 had been accredited for use in New Zealand by International Accreditation New Zealand (IANZ).Separately, Air New Zealand staff were working with the Institute of Environmental Science and Research (ESR) to work out the effectiveness of saliva tests detecting Covid-19.The Rako Science saliva deployment had no connection to trials being run by ESR.Last month, Covid-19 Response Minister Chris Hipkins said Covid-19 the role and effectiveness of saliva testing was still evolving."The Ministry of Health will report back with its findings about the testing in early March," he said at the time. 

COMPARING SURVEILLANCE APPROACHES TO SUPPORT REGAINING FREE STATUS AFTER A FOOT AND MOUTH DISEASE OUTBREAKIntroductionFollowing an FMD eradication program, surveillance will be required to demonstrate that the program has been successful. The World Animal Health Organization (OIE) provides guidelines including waiting periods and appropriate surveillance to support regaining FMD-free status. Serological surveillance is the recommended method for demonstrating freedom but is time consuming and expensive. New technologies such as real-time reverse transcription polymerase chain reaction (RT-qPCR) tests and sampling techniques such as bulk milk testing (BMT) of dairy cattle, oral swabs, and saliva collection with rope tethers in piggeries could enable surveillance to be done more efficiently. Materials and methodsEpidemiological modelling was used to simulate FMD outbreaks, with and without emergency vaccination as part of the response, in Australia. Baseline post-outbreak surveillance approaches for unvaccinated and vaccinated animals based on the European FMD directive (EU 2003) were compared with alternate approaches in which the sampling regime, sampling approaches and/or the diagnostic tests used were varied. The approaches were compared in terms of the resources, time taken, cost, and effectiveness i.e. ability of the surveillance regime to correctly identify the infection status of herds.ResultsIn the non-vaccination scenarios, the alternative approach took less time to compete and cost less, with the greatest benefits seen with larger outbreaks. In vaccinated populations, the alternate surveillance approaches significantly reduced the numbers of herds sampled, the total number of tests done and costs of the post-outbreak surveillance. There was no reduction in effectiveness using the alternate approaches, with one of the benefits being a reduction in the number of false positive herds.DiscussionAlternate approaches to FMD surveillance based on non-invasive sampling methods and RT-qPCR tests have the potential to enable post outbreak surveillance substantiating FMD freedom to be done more quickly and less expensively than traditional approaches based on serological surveys.MG Garner1, W Vosloo2, S. Tapsuwan1, R Bradhurst3, A Seitzinger1, A Breed4 and T Capon11CSIRO-Land and Water, North Road, Acton, 2601, ACT. Australia2CSIRO-Australian Centre for Disease Preparedness, 5 Portarlington Road, 3220, Geelong, Australia3 Centre of Excellence for Biosecurity Risk Analysis, School of BioSciences, University ofMelbourne, Parkville, 3010. Victoria, Australia4Department of Agriculture, Water Resources and the Environment, Canberra. 2601, ACT. Australia

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.11.16.384354v1?rss=1 Authors: Geng, J., Liu, G., Wu, Y., Kong, F., Ma, S., Fu, L. Abstract: Multiple sclerosis (MS) is a complex, progressive neuroinflammatory disease associated with autoimmunity. Currently, effective therapeutic strategy was poorly found in MS. Experimental autoimmune encephalomyelitis (EAE) is widely used to study the pathogenesis of MS. Previous studies have shown that bone marrow mesenchymal stem Cells (BMSCs) transplantation could treat EAE animal models, but the mechanism was divergent. Here, we systematically evaluated whether BMSCs can differentiate into neurons, astrocytes and oligodendrocytes to alleviate the symptoms of EAE mice. We used Immunofluorescence staining to detect MAP-2 neurons marker, GFAP astrocytes marker, and MBP oligodendrocytes marker expression to evaluate whether BMSCs can differentiate. The effect of BMSCs transplantation on inflammatory cell invasion and demyelination in EAE mice were detected by Hematoxylin-Eosin (H&E) and Luxol Fast Blue (LFB) staining. Inflammatory factors expression was detected by ELISA and RT-qPCR. Our results showed that BMSCs could be induced to differentiate into neuron cells, astrocytes and oligodendrocyte in vivo and in vitro. In addition, BMSCs transplant improved the survival rate and weight, and reduced neurological function scores and disease incidence of EAE mice. Moreover, BMSCs transplant alleviated the inflammation and demyelination of EAE mice. Finally, we found that BMSCs transplantation down-regulated the expression levels of pro-inflammatory factors TNF-, IL-1{beta} and IFN-{gamma}, and up-regulated the expression levels of anti-inflammatory factors IL-10 and TGF-{beta}. In conclusion, this study found that BMSCs could alleviate the inflammatory response and demyelination in EAE mice, which may be achieved by the differentiation of BMSCs into neurons, astrocytes and oligodendrocytes in EAE mice. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.14.336982v1?rss=1 Authors: Liu, Z., Ding, H., She, J., Chen, C., Zhang, W.-G., Yang, E. Abstract: Circular RNAs (circRNAs) are involved in various biological processes and in disease pathogenesis. However, only a small number of functional circRNAs have been identified among hundreds of thousands of circRNA species, partly because most current methods are based on circular junction counts and overlook the fact that circRNA is formed from the host gene by back-splicing (BS). To distinguish between expression originating from BS and that from the host gene, we present DEBKS, a software program to streamline the discovery of differential BS between two rRNA-depleted RNA sequencing (RNA-seq) sample groups. By applying real and simulated data and employing RT-qPCR for validation, we demonstrate that DEBKS is efficient and accurate in detecting circRNAs with differential BS events between paired and unpaired sample groups. DEBKS is available at https://github.com/yangence/DEBKS as open-source software. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.05.326645v1?rss=1 Authors: Finetti, L., Pezzi, M., Civolani, S., Calo, G., Scapoli, C., Bernacchia, G. Abstract: In insects, the tyramine receptor 1 (TAR1) has been shown to control several physiological functions, including olfaction. We investigated the molecular and functional profile of the Halyomorpha halys type 1 tyramine receptor gene (HhTAR1) and its role in olfactory functions of this pest. Molecular and pharmacological analyses confirmed that the HhTAR1 gene codes for a true TAR1. The RT-qPCR analysis revealed that HhTAR1 is expressed mostly in adult brain and antennae as well as in early development stages (eggs, 1st and 2nd instar nymphs). In particular, among the antennomeres that compose a typical H. halys antenna, HhTAR1 was more expressed in flagellomeres. Scanning electron microscopy (SEM) investigation revealed the type and distribution of sensilla on adult H. halys antennae: both flagellomeres appear rich in trichoid and grooved sensilla, known to be associated with olfactory functions. Through a RNAi approach, topically delivered HhTAR1 dsRNA induced a 50 % gene downregulation after 24 h in H. halys 2nd instar nymphs. An innovative behavioral assay revealed that HhTAR1 RNAi-silenced 2nd instar nymphs were less susceptible to the alarm pheromone component (E)-2 decenal as compared to control. These results provide critical information concerning the TAR1 role in olfaction regulation, especially alarm pheromone reception, in H. halys. Furthermore, considering the emerging role of TAR1 as target of biopesticides, this work paves the way for further investigation on innovative methods for controlling H. halys. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.31.276253v1?rss=1 Authors: Blackwood, C. A., McCoy, M. T., Ladenheim, B., Cadet, J. L. Abstract: To identify signaling pathways activated by oxycodone self-administration (SA), Sprague-Dawley rats self-administered oxycodone for 20 days using short-access (ShA, 3 h) and long-access (LgA, 9 h) paradigms. Animals were euthanized two hours after SA cessation and dorsal striata were used in post-mortem molecular analyses. LgA rats escalated their oxycodone intake and separated into lower (LgA-L) or higher (LgA-H) oxycodone takers. LgA-H rats showed increased striatal protein phosphorylation of ERK1/2 and MSK1/2. Histone H3, phosphorylated at serine 10 and acetylated at lysine 14 (H3S10pK14Ac), a MSK1/2 target, showed increased abundance only in LgA-H rats. RT-qPCR analyses revealed increased AMPA receptor subunits, GluA2 and GluA3 mRNAs in the LgA-H rats. GluA3, but not GluA2, expression correlated positively with changes in pMSK1/2 and H3S10pK14Ac. Our findings indicate that escalated oxycodone SA results in MSK1/2-dependent histone phosphorylation, which promoted increases in striatal gene expression. Our observations offer novel avenues for pharmacological interventions against oxycodone addiction. Copy rights belong to original authors. Visit the link for more info

Al igual que un pesimista es un optimista bien informado... Un conspiranoico es un oficialnoico realmente bien informado.' Por ello te invitamos a que sepas porque un kit de caldo para paella no puede ser lo que nos traiga esta terrible dictadura pandémica. ………………………………………………………………………………………. Enlaces citados en el podcast: Reacción en cadena de la polimerasa https://es.wikipedia.org/wiki/Reacci%C3%B3n_en_cadena_de_la_polimerasa Precio de los test PCR https://www.biosearchtech.com/products/pcr-kits-and-reagents/pathogen-detection/2019-ncov-cdc-probe-and-primer-kit-for-sars-cov-2 Mascarillas y Bacterias https://www.youtube.com/watch?v=yzJKIVM9AGE& Mascarillas y Bacterias parte 3 https://www.youtube.com/watch?v=o-jwWjqb4_M& Muere Kary Mullis, el Nobel que negaba la existencia del virus del sida y del cambio climático https://elpais.com/elpais/2019/08/13/ciencia/1565717605_558894.html Tuit de Nozick donde Kari Mullis explica en su libro autobiografico como fue expulsado sin poder exponer nada de la conferencia de Toledo https://twitter.com/nozick1/status/1282386129371758592 Christian Drosten. Virólogo que empezó a utilizar las pruebas PCR para el sarcov2 https://en.wikipedia.org/wiki/Christian_Drosten Manual para dummies ¿por qué el test PCR no sirve en absoluto para detectar el Covid? https://www.burbuja.info/inmobiliaria/threads/manual-para-dummies-por-que-el-test-pcr-no-sirve-en-absoluto-para-detectar-el-covid.1331965/ El Dr. Kary Mullis Premio Nobel de Química 1993 dice que el PCR no detecta carga viral https://videos.utahgunexchange.com/watch/el-dr-kary-mullis-premio-nobel-de-quimica-1993-dice-que-el-pcr-no-detecta-carga-viral-mp4_JgZTprRTP4HpbS2.html Sida- La duda. Desinformación y ocultación de los medios (Youtube) https://www.youtube.com/watch?v=3G4_8Ds2b84& Sida- La duda. Desinformación y ocultación de los medios https://videos.utahgunexchange.com/watch/sida-la-duda-desinformacio-n-y-ocultacio-n-de-los-medios-mp4_DWU6fWvxBqg8prP.html Los laboratorios de los CDC fueron contaminados, retrasando las pruebas de coronavirus, dicen los funcionarios https://www.nytimes.com/2020/04/18/health/cdc-coronavirus-lab-contamination-testing.html Hilo sobre test contaminados PCR https://twitter.com/tecn_preocupado/status/1253330556789460993 Tricare militar accidentalmente le dijo a más de 600,000 personas que habían tenido coronavirus https://americanmilitarynews.com/2020/07/military-tricare-accidentally-told-600000-people-theyve-had-coronavirus/ Posible causa de la pandemia por coronavirus: Interferencia inmunológica entre el POLISORBATO 80 de la vacuna antigripal adyuvada y el SARS-CoV-2 https://twitter.com/tecn_preocupado/status/1275825671508496386 El Dr Cesar Carballo dice que muestras congeladas del 2017 daban positivo a la prueba PCR https://twitter.com/LEVANNTE/status/1288763776343461888 Pacientes con COVID = Paciente con Hemoparasitos. https://twitter.com/EliteMonarca/status/1287012229473337345 DEL PANICO Y LA NEGLIGENCIA A LA INVERSION EN SEGURIDAD SANITARIA. FINANCIACION PARA LA PREPARACION ANTE UNA PANDEMIA A NIVEL NACIONAL (2017/2018) https://desmontandoababylon.com/2020/05/03/del-panico-y-la-negligencia-a-la-inversion-en-seguridad-sanitaria-preparacion-ante-futuras-pandemias-a-nivel-nacional/ Event 201, simulacro de coronavirus La parte dedicada al tratamiento en los medios https://desiluminate.org/2020/04/20/event-201-simulacro-de-coronavirus-la-parte-dedicada-al-tratamiento-en-los-medios/ Memorando del Banco Mundial: Desde el Pánico y la Negligencia a la inversión en seguridad sanitaria. (Blog Maria Desiluminate) https://desiluminate.org/2020/06/04/memorando-del-banco-mundial-desde-el-panico-y-la-negligencia-a-la-inversion-en-seguridad-sanitaria/ Científicos de la Autónoma y UTalca publican artículo en prestigiosa revista Nature Biotechnology https://www.uautonoma.cl/news/cientificos-de-la-autonoma-y-utalca-publican-articulo-en-prestigiosa-revista-nature-biotechnology/ Dr. Cesar Carballo muestras congeladas 2017 prueba positiva PCR https://videos.utahgunexchange.com/watch/dr-cesar-carballo-muestras-congeladas-2017-positivo-prueba-pcr-mp4_qhMuf4pdYyHg46d.html ¿Fue la prueba COVID-19 destinada a detectar un virus? https://uncoverdc.com/2020/04/07/was-the-covid-19-test-meant-to-detect-a-virus/ Las pruebas de PCR COVID19 carecen de sentido científico (articulo muy bueno alojado en una página llamada conspiranoica) https://off-guardian.org/2020/06/27/covid19-pcr-tests-are-scientifically-meaningless/ Las mentiras diagnósticas del Coronavirus https://tecnicopreocupado.com/2020/07/08/las-mentiras-diagnosticas-del-coronavirus/ Ministerio de Sanidad, Consumo y Bienestar Social. Estudio Nacional de sero-Epidemiología de la Infección por SARS-CoV-2 en España (ENE-Covid) Preguntas y respuestas frecuentes (pregunta 5 y 6) https://www.mscbs.gob.es/ciudadanos/ene-covid/faqs.htm ¿Me va a dar el resultado de la prueba por escrito? Sí, le daremos el resultado del test rápido de anticuerpos por escrito. Sin embargo, es importante que sepa que este test no permite diagnosticar si usted tiene COVID-19. Este test no diagnostica la enfermedad. Haber tenido contacto con el coronavirus no significa estar enfermo. Tampoco nos dice si usted es contagioso o no, es decir, con este resultado no sabemos si usted puede trasmitir o no el virus a otras personas. Su resultado (positivo o negativo) será útil cuando, en un análisis estadístico posterior, se combine con el de otros participantes para tener datos sobre el grado de contacto de la población española con el virus. ¿El test que me van a hacer es fiable? EL COVID19 es una enfermedad nueva y estos tests que miden si ha habido contacto previo con el virus se han desarrollado recientemente y por tanto la información de cómo funcionan se acaba de generar y no existen por el momento grandes estudios de validación, como nos gustaría. El test tiene el certificado de la Comunidad Europea. No obstante, antes de iniciar ENE-COVID19 (este estudio), hemos probado esta herramienta para confirmar que funciona bien, es decir, que detecta si están presentes los anticuerpos que demuestran que ha habido un contacto anterior con el virus. Se ha probado en alrededor de 100 personas que ya pasaron la enfermedad y en muestras donadas que se utilizaron para diagnosticar otras enfermedades y se habían obtenido antes de que el coronavirus estuviese aquí. Los resultados obtenidos muestran que el test tiene alta sensibilidad y especificidad (mayores al 80%), es decir detecta bastante bien el contacto con el virus. No obstante, necesitamos más información y por eso en este estudio vamos a recoger otros tipos de muestra para poder comparar, cuando dispongamos de mejores herramientas. Es importante que tenga en cuenta que medir el contacto con el coronavirus no es suficiente para diagnosticar COVID19 y tampoco nos dice si usted es o no contagioso en este momento. La fe en la prueba rápida conduce a una epidemia que no era https://www.nytimes.com/2007/01/22/health/22whoop.html María José Martínez Catedrática univ Murcia https://videos.utahgunexchange.com/watch/mari-a-jose-marti-nez-catedra-tica-univ-murcia-pruebas-pcr-mp4_3L3AqYnX2Npnxw4.html Test PCR detecta exosomas https://videos.utahgunexchange.com/watch/test-pcr-2-mp4_mnQtYsbOktHc1R6.html Merienda con Luis de Miguel https://www.youtube.com/watch?v=v3jBEqwlGZA& Y en su web se detallan todos los pasos legales administrativos a seguir si nos quieren hacer una prueba y lo que debemos exigir sobre el test en sí mismo. https://www.scabelum.com/post/__pcr Jefe de la OMS dice que ciertos rebrotes de COVID-19 se deben a "jóvenes que bajan la guardia" https://lta.reuters.com/articulo/salud-coronavirus-oms-idLTAKCN24V2YK-OUSLT?taid=5f22feff31ce9a000122a602&utm_campaign=trueAnthem%3A+Trending+Content&utm_medium=trueAnthem&utm_source=twitter COVID-19 sufre modificaciones genéticas a lo largo del tiempo que es necesario seguir. Aunque hay cebadores de la técnica de PCR que pueden no detectar la presencia de COVID-19, como el protocolo requiere la identificación de objetivos múltiples de COVID-19, se reducen las probabilidades de obtener resultados de identificación de la presencia de COVID-19 falsamente negativos. https://www.iacs.es/covid-19-sufre-modificaciones-geneticas-a-lo-largo-del-tiempo-que-es-necesario-seguir-aunque-hay-cebadores-de-la-tecnica-de-pcr-que-pueden-no-detectar-la-presencia-de-covid-19-como-el-protocolo-requ/ Presidente de Tanzania denuncia graves irregularidades en gestión del Coronavirus. https://www.youtube.com/watch?v=6mQMXKrL58k&&feature=emb_logo Test PCR donde sus fabricantes indican que no sirven para diagnosticar Test PCR VIASURE https://open.lbry.com/@tecnico.preocupado:7/IU-NCO212enes0420-rev.01:e Test PCR VITRO https://open.lbry.com/@tecnico.preocupado:7/MAD-003941M_22720_2138_MAD-003941M-FT-ES-Completa-2020-07-16:d Test PCR RealCycler Monotest CORO v.5.2 https://open.lbry.com/@tecnico.preocupado:7/Manual-deinstruccionesCOROv.5.2:d test PCR GSD NovaPrime https://open.lbry.com/pcov603x_6-sprachig_2020-07-14:9 test PCR Canvax https://open.lbry.com/@tecnico.preocupado:7/qMAXSen™-Coronavirus-SARS-CoV-2-RT-qPCR-detection-Kit-E-and-RdRp-gene-assays:a test PCR canvax 1 https://open.lbry.com/@tecnico.preocupado:7/qMAXSen™-Coronavirus-SARS-CoV-2-RT-qPCR-Detection-Kit-N-and-RNAse-P-genes-based-Assay:5 test PCR canvax 2 https://open.lbry.com/@tecnico.preocupado:7/qMAXSen™-Coronavirus-SARS-CoV-2-RT-qPCR-Detection-Kit-RdRp-gene-assays:d test PCR Viasure S gene https://open.lbry.com/@tecnico.preocupado:7/VS-NCO124enes0420-rev-02:4 Asistentes a discotecas valencianas con brotes de covid-19 se niegan a hacerse test https://www.elconfidencial.com/espana/comunidad-valenciana/2020-07-30/asistentes-discotecas-con-brotes-se-niegan-a-hacerse-test_2701031/ Programa 46 con Maria Jort (Maria desilumínate) donde hablamos sobre la realidad oculta de la pandemia https://www.ivoox.com/programa-46-maria-jort-maria-desiluminate-donde-audios-mp3_rf_50920792_1.html Links y papers de información pruebas pcr: https://www.sciencemag.org/news/2020/02/united-states-badly-bungled-coronavirus-testing-things-may-soon-improve https://www.fda.gov/media/134922/download https://www.sciencedirect.com/topics/immunology-and-microbiology/virus-nucleocapsid https://edition.cnn.com/2020/03/21/politics/us-coronavirus-tests-invs/index.html https://www.cdc.gov/media/releases/2020/t0212-cdc-telebriefing-transcript.html https://www.the-scientist.com/news-opinion/how-sars-cov-2-tests-work-and-whats-next-in-covid-19-diagnostics-67210 https://www.who.int/emergencies/diseases/novel-coronavirus-2019/technical-guidance/laboratory-guidance https://www.npr.org/sections/health-shots/2020/03/26/822084429/in-defense-of-coronavirus-testing-strategy-administration-cited-retracted-study https://www.ncbi.nlm.nih.gov/pubmed/15094372 https://www.dailymail.co.uk/news/article-8171223/Britains-young-coronavirus-victims-Teen-18-youngest-casualty-RSPCA-worker-26-dies.html https://www.spectator.co.uk/article/how-to-understand-and-report-figures-for-covid-19-deaths- https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/877302/guidance-for-doctors-completing-medical-certificates-of-cause-of-death-covid-19.pdf https://www.ons.gov.uk/peoplepopulationandcommunity/birthsdeathsandmarriages/deaths/bulletins/deathsregisteredweeklyinenglandandwalesprovisional/weekending20march2020 https://www.hiddensyria.com/wp-content/uploads/2020/04/COVID-19-Survellance-Bulletin-02.04.20.pdf https://www.cdc.gov/nchs/data/nvss/vsrg/vsrg03-508.pdf https://www.cdc.gov/nchs/data/nvss/coronavirus/Alert-1-Guidance-for-Certifying-COVID-19-Deaths.pdf https://swprs.org/rki-relativiert-corona-todesfaelle/ https://www.faz.net/aktuell/gesellschaft/gesundheit/coronavirus/neue-corona-symptome-entdeckt-virologe-hendrik-streeck-zum-virus-16681450.html?printPagedArticle=true https://www.thecollegefix.com/stanford-epidemiologist-warns-that-coronavirus-crackdown-is-based-on-bad-data/ Los kits de GENOMICA para la detección del Coronavirus Covid-19, listos para comercializarse. https://www.youtube.com/watch?v=u-7pjV0gjao& Toma de muestras (explicación) https://www.youtube.com/watch?v=o6cb6uNyGSU& La leyenda del tablero de ajedrez y los granos de trigo https://matematicascercanas.com/2014/03/10/la-leyenda-del-tablero-de-ajedrez-y-los-granos-de-trigo/ Las Vacunas contra el coronavirus han fallado durante décadas https://cienciaysaludnatural.com/las-vacunas-contra-el-coronavirus-han-fallado-durante-decadas/ ………………………………………………………………..……………………………………………………………….. Música utilizada en este podcast: Tema inicial Start Again by Alex (c) copyright 2011 Licensed under a Creative Commons Attribution (3.0) license. http://dig.ccmixter.org/files/AlexBeroza/31670 Ft: Snowflake & Subliminal LA NUEVA NORMALIDAD - LOS MORANCOS (PARODIA) https://www.youtube.com/watch?v=nez37657-6c&&feature=youtu.be Def con dos - Habrá que morirse más (Videoclip oficial) https://www.youtube.com/watch?v=wmwiPWHpFA0&&feature=youtu.be Rumores de la Caleta by Isaac Albeniz - Cordoba Rodriguez from the Master Series https://www.youtube.com/watch?v=tAUxZFsAmww&&feature=youtu.be Ska-p Se acabo Con Letra https://www.youtube.com/watch?v=1SDIBGFbAMk&&feature=youtu.be JIMBOMAN Piénsalo NO HAY BULLET 2011 https://www.youtube.com/watch?v=euJovbUZGEA&&feature=youtu.be El Niagara En Bicicleta https://www.youtube.com/watch?v=xkqqaIl_vHI&&feature=youtu.be Residente - Apocalíptico (Audio) https://www.youtube.com/watch?v=6_PriJPSr-Q&&feature=youtu.be BUNBURY - Despierta (videoclip) https://www.youtube.com/watch?v=Ig15W89WE-w& Medina Azahara - Necesito Respirar (Lyric Video Oficial) https://www.youtube.com/watch?v=hd3EMefTk7w&&feature=youtu.be ……………………………………………………………….. Epílogo Barón Rojo Mil Años luz. https://www.youtube.com/watch?v=W8yybr9J4II&

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.06.239111v1?rss=1 Authors: Blanco, I., Cabello, J., Urdanoz-Casado, A., Acha, B., Gomez-Orte, E. M., Roldan, M., Perez-Rodriguez, D. R., Mendioroz, M. Abstract: Adult hippocampal neurogenesis (AHN) study is still a challenge. In addition to methodological difficulties is the controversy of results derived of human or animal system approaches. In view of the proven link between AHN and learning and memory impairment, we generated a straightforward in vitro model to recapitulate adult neurogenesis in the context of Alzheimer's disease (AD). Neural progenitor cells (NPCs) monolayer culture was differentiated for a period of 29 days and A{beta} peptide 1-42 was administered once a week. mRNA expression of NEUROD1, NCAM1, TUBB3, RBFOX3, CALB1 and GFAP genes was determined by RT-qPCR. Phenotypic changes were observed during directed differentiation. Except for GFAP and CALB1, these changes correlated with altered expression profile of all genes since 9 days. Only TUBB3 expression remained constant while NEUROD1, NCAM1 and RBFOX3 expression increased over time. Moreover, A{beta} treated NPCs showed transient decreases of mRNA expression for NCAM1, TUBB3 and RBFOX3 genes at 9 or 19 days. Our in vitro human NPCs model is framed within the multistep process of AHN in the SGZ of the DG. Remarkably, its transcriptional assessment might reflect alterations detected in AD human patients, deepening our understanding of the disorder and possibly of its pathogenesis. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.30.228890v1?rss=1 Authors: Woronik, A., Shaffer, H. W., Kiontke, K., Laurent, J. M., Zambrano, R., Boeke, J. D., Fitch, D. H. A. Abstract: Due to the sheer number of COVID-19 (coronavirus disease 2019) cases, the prevalence of asymptomatic cases and the fact that undocumented cases appear to be significant for transmission of the causal virus, SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), there is an urgent need for increased SARS-CoV-2 testing capability that is both efficient and effective. In response to the growing threat of the COVID-19 pandemic in February, 2020, the FDA (US Food and Drug Administration) began issuing Emergency Use Authorizations (EUAs) to laboratories and commercial manufacturers for the development and implementation of diagnostic tests. So far, the gold standard assay for SARS-CoV-2 detection is the RT-qPCR (real-time quantitative polymerase chain reaction) test. However, the authorized RT-qPCR test protocols vary widely, not only in the reagents, controls, and instruments they use, but also in the SARS-CoV-2 genes they target, what results constitute a positive SARS-CoV-2 infection, and their limit of detection (LoD). The FDA has provided a web site that lists most of the tests that have been issued EUAs, along with links to the authorization letters and summary documents describing these tests. However, it is very challenging to use this site to compare or replicate these tests for a variety of reasons. First, at least 12 of 18 tests that were issued EUAs prior to March 31, 2020, are not listed there. Second, the data are not standardized and are only provided as longhand prose in the summary documents. Third, some details (e.g. primer sequences) are absent from several of the test descriptions. Fourth, for tests provided by commercial manufacturers, summary documents are completely missing. To address at least the first three issues, we have developed a database, EUAdb (EUAdb.org), that holds standardized information about EUA-issued tests and is focused on RT-qPCR diagnostic tests, or "high complexity molecular-based laboratory developed tests". By providing a standardized ontology and curated data in a relational architecture, we seek to facilitate comparability and reproducibility, with the ultimate goal of consistent, universal and high-quality testing nationwide. Here, we document the basics of the EUAdb data architecture and simple data queries. The source files can be provided to anyone who wants to modify the database for his/her own research purposes. We ask that the original source of the files be made clear and that the database not be used in its original or modified forms for commercial purposes. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.06.06.138206v1?rss=1 Authors: Meerschaert, K. A., Adelman, P. C., Friedman, R. L., Albers, K. M., Koerber, H. R., Davis, B. M. Abstract: Visceral organs receive neural innervation from sensory ganglia located adjacent to multiple levels of the brainstem and spinal cord. Here we examined whether molecular profiling could be used to identify functional clusters of colon afferents from thoracolumbar (TL), lumbosacral (LS), and nodose ganglia (NG) in the mouse. Profiling of TL and LS bladder afferents was also done. Visceral afferents were back-labeled using retrograde tracers injected into proximal and distal regions of colon or bladder, followed by single cell RT-qPCR and analysis via an automated hierarchical clustering method. Genes were chosen for assay (32 for bladder; 48 for colon) based on their established role in stimulus detection, regulation of sensitivity/function or neuroimmune interaction. A total of 132 colon afferents (from NG, TL and LS) and 128 bladder afferents (from TL and LS) were analyzed. Retrograde labeling from the colon showed NG and TL afferents innervate proximal and distal regions of the colon whereas 98% of LS afferents only project to distal regions. There were clusters of colon and bladder afferents, defined by mRNA profiling, that localized to either TL or LS ganglia. Mixed TL/LS clustering also was found. In addition, transcriptionally, NG colon afferents were almost completely segregated from colon DRG (TL or LS) neurons. These results indicate that populations of primary visceral afferents are functionally ''tuned'' to detect and interact with the internal environment and that information from all levels is integrated at higher (CNS) levels, not only for regulation of homeostatic functions, but for conscious visceral sensations including pain. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.05.31.126227v1?rss=1 Authors: Li, Q., Li, Q.-Q., Jia, J.-N., Liu, Z.-Q., Zhou, H.-H., Jin, W.-L., Mao, X.-Y. Abstract: Background and purpose: Epilepsy is a chronic neurological disease that is characterized by repetitive seizures. Seizures-related complications such as cognitive deficits, anxiety and sleep disorders seriously impact the life quality of patients. Antiepileptic drugs are widely used for the treatment of epilepsy. Sodium valproate is served as the first-line antiepileptic drugs and possesses various pharmacological effects on the brain. Sodium valproate exerts neuroprotective effects in acute nervous system diseases such as ischemic brain damage by inhibiting oxidative stress. However, the mechanism of neuroprotection of sodium valproate in epilepsy is unclear. Lysyl oxidase (Lox) is a monoamine oxidase that acts on extracellular matrix collagen and elastin and it can promote accumulation of oxidative stress. Our previous studies have confirmed that Lox is involved in ferroptosis, a novel iron-dependent and lipid peroxidation-mediated cell death pathway, during epilepsy. In this study, we would like to investigate whether sodium valproate can exert neuroprotective effects on kainic acid-induced epileptic seizures by inhibiting Lox-mediated ferroptosis. Methods: Epileptic mouse models were established by intracranial injection of 250 ng/l kainic acid on right hippocampus. Sodium valproate and ferroptosis inhibitors were administrated by intraperitoneal injecting. The epileptic behavior of the mice within 4 hours was recorded after intracranial injection of kainic acid. Mouse hippocampus was acquired to analyze the mRNA expression of prostaglandin-endoperoxide synthase 2 (PTGS2) and the production of 4-hydroxynonenal (4-HNE). In vitro, the protective effects of sodium valproate on glutamate-induced HT22 cell damage model was assessed by PI/Hoechst staining; The levels of PTGS2, 4-HNE and lipid ROS were analyzed by RT-qPCR, western blot and flow cytometry, respectively. RT-qPCR and Western blot analysis the mRNA and protein expression of Lox in the glutamate-induced HT22 cell damage model. The Lox overexpression model was established by intracranial injection of AAV on right hippocampus. Results: Pretreatment with sodium valproate and ferroptosis inhibitors could significantly alleviate the epileptic seizures in the kainic acid induced epilepsy mouse model. Western blot and RT-qPCR results showed that sodium valproate and ferroptosis inhibitors significantly inhibited the levels of 4-HNE and PTGS2. PI/Hoechst staining showed that 1 mM sodium valproate exerted protective effect on glutamate-induced HT22 cell injury model. There was no significant difference observed between sodium valproate and ferroptosis inhibitors co-intervention group and sodium valproate intervention group on glutamate-induced cell injury model. And sodium valproate could significantly inhibit the production of lipid reactive oxygen species and 4-HNE. The expression of Lox was significantly increased in the glutamate-induced HT22 cell injury model, which could be reversed by pretreatment of sodium valproate. And {beta}-aminopropionitrile (a specific inhibitor of Lox) could inhibit ferroptosis induced by glutamate, as well as ameliorate the epileptic seizures in the kainic acid induced epilepsy mouse model. Pretreatment with sodium valproate could not ameliorate the epileptic behavior in the Lox-overexpression mice. Western blot analysis showed that sodium valproate could not suppress the production of 4-HNE in kainic acid induced epileptic mice model. Conclusions: The neuroprotective effect of sodium valproate in epileptic seizures is closely related to the inhibition of ferroptosis. The inhibition of ferroptosis is involved in the neuroprotective effect of sodium valproate on glutamate-induced HT22 cell damage model. Sodium valproate may exert neuroprotective effects in kainic acid-induced epileptic seizures by abrogating Lox-mediated ferroptosis. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.05.28.120733v1?rss=1 Authors: Jia, J.-N., Li, Q., Sun, Q.-Y., Yang, N., Chen, K.-N., Yin, X.-X., Zhou, H.-H., Mao, X.-Y. Abstract: Background and Purpose: Repetitive epileptic seizures trigger massive neuronal death. Therefore, neuroprotection plays a role in preventing neuronal death and inversely suppresses seizure generation. Additionally, some studies have shown ferroptosis, featured by lipid peroxidation (a dominant form of oxidative stress in the brain), is of paramount importance in epileptic seizures. Lapatinib can play a first-line anti-tumor role by targeting oxidative stress and a recent work illustrates the improvement of encephalomyelitis in rodent models after lapatinib treatment. We hypothesize whether lapatinib can protect against ferroptosis in epileptic seizures via regulating lipid peroxidation. Experimental Approach: The epileptic behavior of the mice was recorded after intracranial injection of KA. Western blot and RT-qPCR were used to detect the protein expression of 4-hydroxynonenal (4-HNE) and glutathione peroxidase 4 (GPX4) and the mRNA expression of prostaglandin endoperoxide synthase 2 (PTGS2) in vivo and in vitro. The level of lipid reactive oxygen species (lipid ROS) in cells pretreated with lapatinib was analyzed by flow cytometry. Key Results: Lapatinib remarkably prevented KA-induced epileptic seizures in mice and ferroptosis was involved in the neuroprotection of lapatinib. Compared with the model group, western blot showed that lapatinib significantly upregulated the levels of GPX4. In the ferroptotic cell death model, lapatinib exerted neuroprotection via up-regulating GPX4. Treatment with Ras-selective lethal small molecule 3 (RSL3), a selective GPX4 inhibitor abrogated its anti-ferroptotic potential. Conclusions and Implications: These results illustrated that lapatinib has neuroprotective potential against KA-triggered epileptic seizures via suppressing GPX4-dependent ferroptosis. Keywords: GPX4; lapatinib; lipid peroxidation; ferroptosis; neuroprotection; epileptic seizures Copy rights belong to original authors. Visit the link for more info

PCR(ポリメラーゼ連鎖反応)の原理について基礎から話しました。dessanをゲストに迎えました。Show notes PCR(ポリメラーゼ連鎖反応) … 特定のDNAサンプルから目的のDNA配列を増幅させる方法。分子生物学における最も基本的な実験の一つ。 DNA(デオキシリボ核酸) … 地球上の生命体の遺伝情報を司る高分子化学物質。いわゆるDNA。 水素結合 … 水素間で起きる非共有結合性の相互作用。2本のDNA鎖が反平行に配向し二重らせん構造を形成する際、A-T,G-Cの間は水素結合によって結ばれる。A-Tは2個、G-Cは3個の水素結合によって結ばれるため、結合の強度が異なる。共有結合やイオン結合と比べて弱く、ファンデルワールス力よりは強い。 DNAポリメラーゼ … DNAを複製するために必要なタンパク質。普通のタンパク質は高温にすると失活して壊れてしまうため、PCRのようにサンプルを高温状態にするとDNAポリメラーゼは失活してしまう。初期のプロトコルではこのため、毎回DNAポリメラーゼが入れ替えられていたが、高温化で生息する好熱性細菌から耐熱DNAポリメラーゼが発見され、PCRが一気に広まった。 半保存的複製 … 相補的な二本鎖DNAがどのように複製されるのか。DNAが非常に情報メディアとして特有の振る舞いをみせる。DNA複製システムを明らかにしたのはスタールとメセルソン。メセルソンについてはep7を参照のこと。 DNA複製 … 生物が細胞を倍加していく上で、遺伝情報を担うDNAも二倍にする必要がある。 プライマー … DNAポリメラーゼは基本的に一本鎖DNAをそのまま複製することはできず、一旦二本鎖にする必要がある。その時に使われるのがプライマーと呼ばれる短い塩基配列である。生体内では基本短鎖RNAでPCR上では短鎖DNAが用いられる。 サーマルサイクラー … PCRのためにサンプルの温度を上下させるための機械 電気泳動 … 荷電している分子を電場中で移動させる。これによりDNAやタンパク質を分離する方法。 RNAウィルス … 遺伝物質としてRNAを用いているウィルスのこと。 エチジウムブロマイド … 二本鎖DNAに架橋する。紫外線を当てると蛍光を発するため、核酸の検出などに用いられる。 qPCR(定量PCR) RT-qPCR Researchat.fm ep37 … 前回のdessan出演回。分子生物学の基礎について話しています。 Researchat.fm ep24 … PCRの発明者でノーベル賞を受賞したKary Mullis追悼回。こちらでもPCRについて詳しく話しています。 TAKARAのPCRの説明とそのプロトコル … PCRキットを販売するTAKARAのPCRの説明とそのプロトコル。非常にわかりやすい。トラブルシューティングもしっかりと書かれているのでPCRが動かない時はしっかりプロトコルを読もう！特にP.6からのPCRの原理の部分はPCR原理の理解の助けになると思います。 dessanによるPCRまとめ図 Editorial notes 私の部屋で撮ったのでハウスメイトにビビって最初のほうは音声が小さいです。申し訳ありません。(dessan) 生物の機構とPCRの対比を行うことでわかりやすくなったのではないかと期待しております。 (tadasu)

Volume 24 Issue 6, December 2019Dave Pechter discusses with Chao Li the article, "Automated System for Small-Population Single-Particle Processing Enabled by Exclusive Liquid Repellency." Lossless processing and culture of rare cells (e.g., circulating tumor cells, drug-persistent microorganisms) at single-cell level is of great significance in understanding the heterogeneity of carcinogenesis or human pathogenesis caused by microbial infection. Current single-cell isolation techniques like fluorescence-activated cell sorting (FACS) require relatively large sample volume and cell number to work with, and inflict sample loss and reduced cell viability from the processing. While microfluidic and single-cell printing techniques allow the handling of minute amounts of cellular samples, they either come with limited physical access to the sample of interest due to the closed-channel design (e.g., droplet microfluidics) or sample loss during aspiration, transfer, and sample retrieval from culture.Recently, we reported an extreme wettability phenomenon, named exclusive liquid repellency or ELR. ELR is observed in solid-liquid-liquid three phase systems, where a solid surface shows complete repellency to a liquid (with Young's contact angle, CA = 180o) when exposed to a second immiscible liquid. This phenomenon is observed when a particular thermodynamic boundary condition is satisfied (i.e., γS/Lcp + γLdp/Lcp ≤ γS/Ldp, where γ - interfacial tension, S - solid, Lcp - liquid of continuous phase, and Ldp - liquid of dispersed phase). Neither surfactant nor flow condition is required, e.g., compared with droplet microfluidics. ELR enables additional fluidic control, robust on-chip cell culture, and improved processing of rare cell samples in open aqueous fluid under oil. ELR is distinct from other liquid repellent systems with CA < 180o (i.e., non-ELR), showing no compromise of liquid adhesion on solid surfaces and enabling unique applications.In this work, we developed an automated platform using ELR microdrops for lossless single-particle (or single-cell) isolation, identification, and retrieval. It features the combined use of a robotic liquid handler, an automated microscopic imaging system, and real-time image-processing software for single-particle identification. The automated ELR technique enables rapid, hands-free, and robust isolation of microdrop-encapsulated rare cellular samples, and further on-chip cell culture or down-stream analysis (e.g., RNA extraction and RT-qPCR).

Classical and African swine fever belong to the most important contagious diseases of pigs worldwide and are notifiable to the World Organization for Animal Health (OIE). While outbreaks of classical swine fever (CSF) have a long and ongoing history in Europe, up to now African swine fever (ASF) was considered an exotic disease within EU Member States. However, very recently the disease has been introduced into the European wild boar and domestic pig population in Poland, Lithuania, Latvia, and Estland. Hence, both diseases pose a high risk to the whole European pig industry and wildlife. Regardless of very similar clinical pictures that are not discriminable without laboratory diagnosis, the causative agents differ greatly. Classical swine fever is caused by a small enveloped positive single-stranded RNA virus belonging to the genus Pestivirus within the Flaviviridae family. The causative agent of ASF is a complex DNA virus of the genus Asfivirus within the Asfarviridae family representing the only DNA arthropod-borne (ARBO) virus. Classical swine fever virus (CSFV) isolates of recent European outbreaks are characterized by their moderate virulence. The clinical picture can range from an almost inapparent infection to a lethal hemorrhagic fever like illness. High variability in disease course and outcome are a challenge for both disease surveillance and pathogenetic research. Although several studies aimed to analyze basic pathogenetic mechanisms, responsible factors have never been elucidated entirely. While on the host’s side, age and immune status are acknowleged parameters, the virulence of the isolate seems to be decisive on the agent’s side. Moreover, the influence of the genetic background of the host has been discussed. To define host responses linked to different disease courses and outcome, a first animal trial was conducted with a moderately virulent CSFV strain and different pig breeds including European wild boar. In a second trial, the impact of the age was revisited in combination with the assessment of tools for active swine fever surveillance. Dysregulation of immune responses, especially cytokine reactions, seems to play a crucial role in CSF pathogenesis. Up to now, there has been a serious lack of appropriate and reliable tools for cytokine gene expression analyses, especially in pigs. To overcome this shortcoming, a harmonized TaqMan-based RT-qPCR protocol for the detection of seven swine fever relevant cytokines was developed and fully validated. This assay is now available for future studies and could be implemented also for other swine diseases including ASF. Beyond the studies focusing on underlying mechanisms, diagnostic approaches for active and passive disease surveillance in wild boar have been targeted. Sample submission is usually the bottleneck of wildlife surveillance under field conditions, even in times of increased risk. Pragmatic approaches for sampling and transport could improve the compliance of hunters. In view of the fact that an introduction of ASF and CSF is usually accompanied by an increased mortality, animals found dead have to be sampled even if they are already in various stages of decay. To this means, a dry-/ semi-dry blood swab method was implemented enabling an easy handling under field and laboratory conditions. Moreover, a simple approach for sampling of live animals for active surveillance, i.e. a “rope-in-a-bait” method, was evaluated with respect to the frequently observed subclinical CSF forms in older animals.

ATX-II wurde ursprünglich aus dem Gift der Seeanemone Anemonia sulcata isoliert und ist als potenter Aktivator spannungsgesteuerter Natriumionenkanäle (Navs) bekannt, da es einen persistierenden Na+-Strom induziert. Vor Kurzem wurden bestimmte Nav Subtypen mit menschlichem Schmerz in Verbindung gebracht. Somit liegt es nahe, dass detaillierte Kenntnisse über die Regulierung von Navs für die Entwicklung neuer Pharmaka im Bereich der Schmerztherapie von Vorteil sind. Auch der durch einen alternativen Inaktivierungsmechanismus entstehende resurgent current spielt nach Erkenntnissen der jüngsten Zeit eine Rolle im Schmerzgeschehen. Er kann einen Angriffspunkt für neue Analgetika darstellen, so man seine Entstehung und mögliche Modifikationen entschlüsseln kann. Das Ziel dieser Arbeit war es, zu untersuchen, ob ATX-II resurgent currents von Navs in großen und kleinen murinen DRGs verändern bzw. induzieren kann. Zudem sollte beleuchtet werden, ob und wenn ja, welche Wirkung das Toxin auf periphere A- und C Schmerzfasern hat. Die Auswirkungen von ATX-II auf Navs wurden in großen und kleinen DRGs der Maus sowie in Zelllinien mit whole-cell voltage-clamp Messungen untersucht. Gesunde humane Probanden bewerteten Empfindungsänderungen (Schmerz und Juckreiz) nach intrakutaner Injektion von ATX-II. Mit Hilfe des "Laser Doppler Imagers" wurde das mögliche Auftreten eines Axon Reflex Erythems untersucht. Die mittels des FACS Zellsortierers nach Zellgröße sortierten DRGs wurden mit RT-qPCR auf ihren Gehalt an Nav1.6, Nav1.7 und 4 mRNA untersucht. ATX-II verstärkte resurgent currents in großen DRGs, zeigte in kleinen jedoch keinen Effekt. Im heterologen Expressionssystem trat ein resurgent current erst auf, wenn 4 Peptid über die Intrazellulärflüssigkeit zur Verfügung stand. Korrelierend zu den Befunden auf Zellebene rief ATX-II bei intrakutaner Injektion im Menschen nur solange stechenden Schmerz und veränderte mechanische Empfindung hervor, wie die A-Fasern für die Schmerzleitung verfügbar waren. Nachdem ein Nervenkompressionsblock angelegt und die Leitung via A-Fasern blockiert war, waren die ATX-II vermittelten Effekte verschwunden. Zudem konnte kein Axon Reflex Erythem gefunden werden, das ein Zeichen für eine C-Schmerzfaser-Aktivierung wäre. Die Ergebnisse dieser Arbeit legen nahe, dass geringe Konzentrationen von ATX-II eine selektive Wirkung auf große DRGs haben. Erst durch Hinzufügen von 4-Peptid ist es möglich mit dem Toxin resurgent current in kleinen DRGs sowie Zelllinien mit heterolog exprimierten Nav1.6 und Nav1.7 zu induzieren. Dies lässt darauf schließen, dass es kleinen DRGs an 4 in ausreichender Menge mangelt und deshalb kein resurgent current sichtbar ist. Ebenso übt ATX-II einen Effekt auf A Fasern aus, die mit großen DRGs verbunden sind, wohingegen C-Fasern weitgehend unbeeinflusst bleiben. Die intrakutane Injektion von ATX-II führt zu stechendem Schmerz und einer Juckreiz ähnlichen Empfindung, die sich durch A-Faser-Block verhindern lässt. Daraus kann man schließen, dass ATX-II in niedrigen Konzentrationen als spezifischer A-Faser Aktivator fungiert.

Breast cancer is still the most frequent cause of cancer-related death in women worldwide. Often death is not caused only by the primary tumour itself, but also by metastatic lesions. Today it is largely accepted, that these remote metastases arise out of cells, which detach from the primary tumour, enter circulation, settle down at secondary sites in the body and are called Circulating Tumour Cells (CTCs). The occurrence of such minimal residual diseases in the blood of breast cancer patients is mostly linked to a worse prognosis for therapy outcome and overall survival. Due to their very low frequency, the detection of CTCs is, still a technical challenge. RT-qPCR as a highly sensitive method could be an approach for CTC-detection from peripheral blood of breast cancer patients. This assumption is based on the fact that CTCs are of epithelial origin and therefore express a different gene panel than surrounding blood cells. For the technical approach it is necessary to identify appropriate marker genes and to correlate their gene expression levels to the number of tumour cells within a sample in an in vitro approach. After that, samples from adjuvant and metastatic patients can be analysed. This approach may lead to new concepts in diagnosis and treatment

Introduction: This study aimed to characterize the glycophenotype of osteoarthritic cartilage and human chondrocytes. Methods: Articular knee cartilage was obtained from nine osteoarthritis (OA) patients. mRNA levels for 27 glycosyltransferases were analyzed in OA chondrocytes using RT-qPCR. Additionally, N- and O-glycans were quantified using mass-spectrometry. Histologically, two cartilage areas with Mankin scores (MS) either

Allergic asthma has a high prevalence and is characterized by airway inflammation, tissue remodeling and a decline in respiratory function. Although the pathogenesis is well known, the underlying mechanisms are still poorly understood. It is believed that a fine interplay exists between the exposure to environmental stimuli and relatively small changes in expression of several genes with inter-individual variation. As microRNAs (miRNAs) are known to be responsive to environmental exposures and show dysregulated levels in diseased states, their function as regulators of gene expression might be a missing link for the changes seen in asthma. In this project, changes in miRNA expression in a mouse model for allergic asthma were investigated and the interaction with possible target genes was analyzed. Female Balb/c mice were i. p. sensitized with ovalbumin (OVA) followed by aerosol challenge on two consecutive days. An asthmatic phenotype was confirmed by elevated total cell numbers due to a rise in inflammatory cells, as well as increased CCL17 levels in broncho-alveolar lavage (BAL). High titres of OVA-specific serum IgE were measured and lung histopathology revealed infiltration of inflammatory cells with eosinophilia. To study changes in miRNA expression, whole lung RNA was subjected to miRNA-microarray analysis (Exiqon). From 580 screened miRNAs, 319 were found to be expressed, of which 36 were differentially regulated in the allergic asthma group compared to healthy control mice. A second, TaqMan® chemistry based array was performed for validation. Based on the overlap between the two arrays in addition to fold changes and p-values (Exiqon), eight miRNAs were selected for single RT-qPCR measurement. Dysregulated expression of six miRNAs could be confirmed (miRNA-21, -142-3p, -144, -205, -208, -451). Due to relatively low fold changes and in order to monitor possible co-regulation, the top 100 differentially regulated miRNAs from the Exiqon array were included in an in situ target prediction. Applying a “full consensus” approach of five prediction algorithms, 961 putative target genes were identified. Based on the assumption, that target genes harboring multiple miRNA sites might be more relevant, 11 targets containing more than four miRNA binding sites were selected. From these, the transcription factor cAMP-responsive element-binding protein 1 (CREB1) was chosen for further analysis because of its previous association with asthmatic disease. Moreover, four miRNAs (miRNA-17, -22, -144, -181a) were predicted to bind at eight different sites, one of them being miRNA-144, a significantly up-regulated miRNA identified in the initial asthma profile. To experimentally test the functional interaction between CREB1 and the predicted miRNAs, a CREB1 3´-untranslated region (UTR) containing luciferase based reporter plasmid vector was constructed and co-transfected with precursor (pre-) miRNAs into human bronchial epithelial cells. Binding of all four miRNAs could be confirmed by measuring luciferase expression. Furthermore, three of four miRNAs, when transfected alone, were able to down-regulate endogenous CREB1 expression in vitro. In the lung tissue of asthma mice, CREB1 mRNA levels were significantly reduced compared to healthy controls, in contrast to two miRNAs, miRNA-17 and -144, which showed up-regulation. To gain further insight into expression patterns during sensitization and after challenge, expression of CREB1, the two validated binding partners miRNA-17, and -144, as well as the two miRNAs (miRNA-21, -451) with most significant p-values and high fold changes from the Exiqon array were analyzed. Clear expression changes happened after OVA aerosol challenge with CREB1 levels being steadily decreased, whereas all tested miRNAs showed elevated levels at 24 h post challenge, which further intensified after 120 h. This increase resembles measurements of inflammatory cell counts in BAL pointing at a possible origin within this population. In order to test whether findings can be translated into the human situation, miRNA changes in whole blood samples of mice were compared to miRNA patterns in peripheral blood of asthmatic children. In contrast to measurements in lung tissue, all four miRNAs showed markedly decreased expression in murine blood. In human samples this reduction was mirrored and significant for miRNA-144 and -451.