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Nina Schaller,1 Helge Krusemark,1 Esther Mende,1 Michael Weiß,1 Bianca Spanier,1 Otto Zelger,1 Jan Bischof,1 Bernhard Haller,2 Martin Halle,1,3,&ast; Monika Siegrist1,&ast; 1Department of Prevention and Sports Medicine, School of Medicine, University Hospital ‘rechts der Isar’, Technical University of Munich, Munich, Germany; 2Institute of AI and Informatics in Medicine, University Hospital ‘rechts der Isar’, Technical University of Munich, Munich, Germany; 3DZHK (German Center for Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany&ast;These authors contributed equally to this workCorrespondence: Nina Schaller, Department of Prevention and Sports Medicine, School of Medicine, University Hospital ‘rechts der Isar’, Technical University of Munich, Georg-Brauchle-Ring 56, Munich, 80992, Germany, Tel +49 89 289 24441, Fax +49 89 289 24451, Email nina.schaller@mri.tum.dePurpose: Mobility is a crucial factor for independence and quality of life in old age. Nevertheless, many old people in retirement homes do not meet the physical activity recommendations. The aim of the Bestform-F – Best Function of Range of Motion feasibility study (bestform-F) was to evaluate the feasibility of implementing a machine-based multimodal exercise training program in older residents in retirement homes.Materials and Methods: The participants (n = 77) were recruited from two retirement homes and took part in a six-month multimodal exercise training program (2x/week, 45 minutes) on pneumatic strength training machines, a balance platform and bicycle ergometers. Feasibility criteria were recruitment number ≥ 35 participants within six months, dropout rate < 40% of participants within six months of exercise, and training adherence ≥ 50% of participants taking part in at least 50% of offered training sessions. Additionally, physical performance, fear of falling, cognitive function, and quality of life were assessed at baseline and after six months.Results: For the bestform-F study, 77 (85.6 ± 6.6 years; 78% women) out of 215 eligible residents from two senior residences were recruited. The dropout rate over six months was 10% (8/77 participants). The training adherence rate for the finishing participants was 77% (53/69 participants). In addition to the achieved feasibility criteria, significant improvements were recorded in the Chair Stand Test, Six-Minute Walk Test, and fear of falling after six months.Conclusion: All feasibility criteria have been fulfilled. The high number of recruited participants, the low dropout rate, and high adherence to the training program confirm the feasibility of a multimodal machine-based exercise training program offered to residents in retirement homes. The results provide a basis for a cluster-randomized controlled trial aimed at further investigating the efficacy of the bestform-F program.Keywords: machine-based exercise, seniors, sarcopenia, mobility, resistance training, balance

A firm biophysical basis for the cryopreservation of human spermatozoa is limited by a lack of knowledge regarding the water permeability characteristics during freezing in the presence of extracellular ice and cryoprotective agents (CPA). Cryomicroscopy cannot be used to measure dehydration during freezing in human spermatozoa because of their highly non-spherical shape and their small dimensions which are at the limits of light microscopic resolution. Using a new shape-independent differential scanning calorimeter (DSC) technique, volumetric shrinkage during freezing of human sperm cell suspensions was obtained at cooling rates of 5 and 10 degrees C/min in the presence of extracellular ice and CPA. Using previously published data, the human sperm cell was modelled as a cylinder of length 40.2 micrometer and a radius of 0.42 micrometer with an osmotically inactive cell volume, V(b), of 0.23V(o), where V(o) is the isotonic cell volume. By fitting a model of water transport to the experimentally obtained volumetric shrinkage data, the best fit membrane permeability parameters (L(pg) and E(Lp)) were determined. The 'combined best fit' membrane permeability parameters at 5 and 10 degrees C/min for human sperm cells in modified media are: L(pg) = 2. 4x10(-14) m(3)/Ns (0.14 micrometer/min-atm) and E(Lp) = 357.7 kJ/mol (85. 5 kcal/mol) (R(2) = 0.98), and in CPA media (with 6% glycerol and 10% egg yolk) are L(pg)[cpa] = 0.67x10(-14) m(3)/Ns (0.04 micrometer/min-atm) and E(Lp)[cpa] = 138.9 kJ/mol (33.2 kcal/mol) (R(2) = 0.98). These parameters are significantly different from previously published parameters for human spermatozoa obtained at suprazero temperatures and at subzero temperatures in the absence of extracellular ice. The parameters obtained in this study also suggest that damaging intracellular ice formation (IIF) could occur in human sperm cells at cooling rates as low as 25-45 degrees C/min, depending on the concentrations of the CPA. This may help to explain the discrepancy between the empirically determined optimal cryopreservation cooling rates (<100 degrees C/min) and the numerically predicted optimal cooling rates (>7000 degrees C/min) obtained using previously published suprazero human sperm permeability parameters which do not account for the presence of extracellular ice. [ABSTRACT FROM AUTHOR]

Abstract Background Chondrosarcomas are malignant cartilage tumors that do not respond to traditional chemotherapy or radiation. The 5-year survival rate of histologic grade III chondrosarcoma is less than 30%. An animal model of chondrosarcoma has been established - namely, the Swarm Rat Chondrosarcoma (SRC) - and shown to resemble the human disease. Previous studies with this model revealed that tumor microenvironment could significantly influence chondrosarcoma malignancy. Methods To examine the effect of the microenvironment, SRC tumors were initiated at different transplantation sites. Pyrosequencing assays were utilized to assess the DNA methylation of the tumors, and SAGE libraries were constructed and sequenced to determine the gene expression profiles of the tumors. Based on the gene expression analysis, subsequent functional assays were designed to determine the relevancy of the specific genes in the development and progression of the SRC. Results The site of transplantation had a significant impact on the epigenetic and gene expression profiles of SRC tumors. Our analyses revealed that SRC tumors were hypomethylated compared to control tissue, and that tumors at each transplantation site had a unique expression profile. Subsequent functional analysis of differentially expressed genes, albeit preliminary, provided some insight into the role that thymosin-β4, c-fos, and CTGF may play in chondrosarcoma development and progression. Conclusion This report describes the first global molecular characterization of the SRC model, and it demonstrates that the tumor microenvironment can induce epigenetic alterations and changes in gene expression in the SRC tumors. We documented changes in gene expression that accompany changes in tumor phenotype, and these gene expression changes provide insight into the pathways that may play a role in the development and progression of chondrosarcoma. Furthermore, specific functional analysis indicates that thymosin-β4 may have a role in chondrosarcoma metastasis.

Abstract Background Proximal chromosome 15q is implicated in neurodevelopmental disorders including Prader-Willi and Angelman syndromes, autistic disorder and developmental abnormalities resulting from chromosomal deletions or duplications. A subset of genes in this region are subject to genomic imprinting, the expression of the gene from only one parental allele. Results We have now identified the NDNL2 (also known as MAGE-G) gene within the 15q autistic disorder susceptibility region and have mapped its murine homolog to the region of conserved synteny near necdin (Ndn) on mouse Chr 7. NDNL2/MAGE-G is a member of a large gene family that includes the X-linked MAGE cluster, MAGED1 (NRAGE), MAGEL2 and NDN, where the latter two genes are implicated in Prader-Willi syndrome. We have now determined that NDNL2/Ndnl2 is widely expressed in mouse and human fetal and adult tissues, and that it is apparently not subject to genomic imprinting by the PWS/AS Imprinting Center. Conclusion Although NDNL2/MAGE-G in the broadly defined chromosome 15 autistic disorder susceptibility region, it is not likely to be pathogenic based on its wide expression pattern and lack of imprinted expression.

Breast cancer is the most common cause of death from cancer in women. Here, we present the case of a 43-year-old woman, who received a diagnosis of claudin-low luminal B breast cancer. The lesion revealed to be a poorly differentiated high-grade infiltrating ductal carcinoma, which was strongly estrogen receptor (ER)/progesterone receptor (PR) positive and human epidermal growth factor receptor (HER2) negative. Her tumor underwent in-depth chromosomal, mutational and gene expression analyses. We found a pathogenic protein truncating mutation in the TP53 gene, which is predicted to disrupt its transcriptional activity. The patient also harbors germline mutations in some mismatch repair (MMR) genes, and her tumor displays the presence of immune infiltrates, high tumor mutational burden (TMB) status and the apolipoprotein B mRNA editing enzyme catalytic polypeptide 3 (APOBEC3) associated signatures, which, overall, are predictive for the use of immunotherapy. Here, we propose promising prognostic indicators as well as potential therapeutic strategies based on the molecular characterization of the tumor., (© 2024. The Author(s).)

Research Infrastructures (RIs) are strategic assets facilitating innovation and knowledge advancement across all scientific disciplines. They provide researchers with advanced tools and resources that go beyond individual or institutional capacities and promote collaboration, community-building and the application of scientific standards. Remote and virtual access to RIs enables scientists to use these essential resources without the necessity of being physically present. The COVID-19 pandemic restrictions where a catalyst for the expansion and further development of remote and virtual access models, particularly in fields where physical access had been the predominant model. The eRImote project explores pathways for digital and remote RI access through targeted surveys, stakeholder workshops, expert groups discussions, and the analysis of specific use cases. This paper provides a definition of remote and virtual access and remote training and explores their implementation across various RIs, highlighting the implications for their operational processes and the dynamics of interaction between RIs and their user communities. It presents the identified advantages, obstacles, and best-practices, alongside strategies and recommendations to navigate and mitigate challenges effectively. Key issues and recommendations are summed up separately for remote access, virtual access, and remote training, complemented by general recommendations for facilitating remote and virtual access to RIs. These relate to budgeting and funding, the balancing of RI access models, the need for regulatory frameworks for sample shipments, collaboration among RIs, impact assessment of remote and virtual access on user interactions, operational efficiency and the environment footprint of RIs, and the adaption of data sharing policies. Stakeholders are broadly invited to give their feedback on the paper's findings and conclusions, which will be integrated into improved versions of this paper., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 RAESS M et al.)

The increasing incidence of urothelial bladder cancer is a notable global concern, as evidenced by the epidemiological data in terms of frequency, distribution, as well as mortality rates. Although numerous molecular alterations have been linked to the occurrence and progression of bladder cancer, currently there is a limited knowledge on the molecular signature able of accurately predicting clinical outcomes. In this report, we present a case of a pT3b high-grade infiltrating urothelial carcinoma with areas of squamous differentiation characterized by very high tumor mutational burden (TMB), with up-regulations of immune checkpoints. The high TMB, along with elevated expressions of PD-L1, PD-L2, and PD1, underscores the rationale for developing a personalized immunotherapy focused on the use of immune-checkpoint inhibitors. Additionally, molecular analysis revealed somatic mutations in several other cancer-related genes, including TP53, TP63 and NOTCH3. Mutations of TP53 and TP63 genes provide mechanistic insights on the molecular mechanisms underlying disease development and progression. Notably, the above-mentioned mutations and the elevated hypoxia score make the targeting of p53 and/or hypoxia related pathways a plausible personalized medicine option for this bladder cancer, particularly in combination with immunotherapy. Our data suggest a requirement for molecular profiling in bladder cancer to possibly select appropriate immune-checkpoint therapy., (© 2024. The Author(s).)

Microinjection is a technique used for transgenesis, mutagenesis, cell labeling, cryopreservation, and in vitro fertilization in multiple single and multicellular organisms. Microinjection requires specialized skills and involves rate-limiting and labor-intensive preparatory steps. Here, we constructed a machine-vision guided generalized robot that fully automates the process of microinjection in fruit fly (Drosophila melanogaster) and zebrafish (Danio rerio) embryos. The robot uses machine learning models trained to detect embryos in images of agar plates and identify specific anatomical locations within each embryo in 3D space using dual view microscopes. The robot then serially performs a microinjection in each detected embryo. We constructed and used three such robots to automatically microinject tens of thousands of Drosophila and zebrafish embryos. We systematically optimized robotic microinjection for each species and performed routine transgenesis with proficiency comparable to highly skilled human practitioners while achieving up to 4× increases in microinjection throughput in Drosophila. The robot was utilized to microinject pools of over 20,000 uniquely barcoded plasmids into 1,713 embryos in 2 days to rapidly generate more than 400 unique transgenic Drosophila lines. This experiment enabled a novel measurement of the number of independent germline integration events per successfully injected embryo. Finally, we showed that robotic microinjection of cryoprotective agents in zebrafish embryos significantly improves vitrification rates and survival of cryopreserved embryos post-thaw as compared to manual microinjection. We anticipate that the robot can be used to carry out microinjection for genome-wide manipulation and cryopreservation at scale in a wide range of organisms., Competing Interests: Conflicts of interest ADA, ASJ, DMG, and SBK are co-founders of Objective Biotechnology Inc. which is commercializing the technology described in this manuscript., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America.)

Renal cell carcinoma originates from the lining of the proximal convoluted renal tubule and represents the most common type of kidney cancer. Risk factors and comorbidities might be associated to renal cell carcinoma, while a small fraction of 2-3% emerges from patients with predisposing cancer syndromes, typically associated to hereditary mutations in VHL, folliculin, fumarate hydratase or MET genes. Here, we report a case of renal cell carcinoma in patient with concurrent germline mutations in BRCA1 and RAD51 genes. This case displays an unusual high mutational burden and chromosomal aberrations compared to the typical profile of renal cell carcinoma. Mutational analysis on whole genome sequencing revealed an enrichment of the MMR2 mutational signature, which is indicative of impaired DNA repair capacity. Overall, the tumor displayed a profile of unusual high genomic instability which suggests a possible origin from germline predisposing mutations in the DNA repair genes BRCA1 and RAD51. While BRCA1 and RAD51 germline mutations are well-characterised in breast and ovarian cancer, their role in renal cell carcinoma is still largely unexplored. The genomic instability detected in this case of renal cell carcinoma, along with the presence of unusual mutations, might offer support to clinicians for the development of patient-tailored therapies., (© 2024. The Author(s).)

Competing Interests: Competing interests statement:U.A.G. and Case Western Reserve University have financial interests in Hemex Health Inc. E.K., U.A.G., and Case Western Reserve University have financial interests in BioChip Labs Inc. U.A.G. and Case Western Reserve University have financial interests in Xatek Inc. U.A.G. has financial interests in DxNow Inc. Financial interests include licensed intellectual property, stock ownership, research funding, employment, and consulting. Hemex Health Inc. offers point-of-care diagnostics for hemoglobin disorders, anemia, and malaria. BioChip Labs Inc. offers commercial clinical microfluidic biomarker assays for inherited or acquired blood disorders. Xatek Inc. offers point-of-care global assays to evaluate the hemostatic process. DxNow Inc. offers microfluidic and bio-imaging technologies for in vitro fertilization, forensics, and diagnostics. Competing interests of Case Western Reserve University employees are overseen and managed by the Conflict of Interests Committee according to a Conflict-of-Interest Management Plan. The study was conducted prior to E.K. employment at IDEXX Laboratories, and the ideas expressed are not in his capacity as an IDEXX employee. A.D. serves on the Scientific Advisory Board of Hemanext Inc. and Macopharma Inc., and his competing interests are managed by the University of Colorado in accordance with their conflict-of-interest policies. S.N.T. and M.T. serve on the Scientific Advisory Board of Sylvatica Biotech Inc., a company focused on developing high subzero organ preservation technology. Competing interests for all authors affiliated with the Massachusetts General Hospital are managed by the MGH and Mass General Brigham in accordance with their conflict-of-interest policies. The remaining authors declare no competing interests.

The monogenetic disease epidermolysis bullosa (EB) is characterised by the formation of extended blisters and lesions on the patient's skin upon minimal mechanical stress. Causal for this severe condition are genetic mutations in genes, leading to the functional impairment, reduction, or absence of the encoded protein within the skin's basement membrane zone connecting the epidermis to the underlying dermis. The major burden of affected families justifies the development of long-lasting and curative therapies operating at the genomic level. The landscape of causal therapies for EB is steadily expanding due to recent breakthroughs in the gene therapy field, providing promising outcomes for patients suffering from this severe disease. Currently, two gene therapeutic approaches show promise for EB. The clinically more advanced gene replacement strategy was successfully applied in severe EB forms, leading to a ground-breaking in vivo gene therapy product named beremagene geperpavec (B-VEC) recently approved from the US Food and Drug Administration (FDA). In addition, the continuous innovations in both designer nucleases and gene editing technologies enable the efficient and potentially safe repair of mutations in EB in a potentially permanent manner, inspiring researchers in the field to define and reach new milestones in the therapy of EB.