Harnessing diversity to study Alzheimer's disease: A new iPSC resource from the NIH CARD and ADNI.

The iDA Project (iPSCs to Study Diversity in Alzheimer's and Alzheimer's Disease-related Dementias) is generating 200 induced pluripotent stem cell lines from Alzheimer's Disease Neuroimaging Initiative participants. These lines are sex balanced, include common APOE genotypes, span disease stages, and are ancestrally diverse. Cell lines and characterization data will be shared openly.

A fully automated FAIMS-DIA mass spectrometry-based proteomic pipeline.

Here, we present a standardized, "off-the-shelf" proteomics pipeline working in a single 96-well plate to achieve deep coverage of cellular proteomes with high throughput and scalability. This integrated pipeline streamlines a fully automated sample preparation platform, a data-independent acquisition (DIA) coupled with high-field asymmetric waveform ion mobility spectrometer (FAIMS) interface, and an optimized library-free DIA database search strategy. Our systematic evaluation of FAIMS-DIA showing single compensation voltage (CV) at -35 V not only yields the deepest proteome coverage but also best correlates with DIA without FAIMS. Our in-depth comparison of direct-DIA database search engines shows that Spectronaut outperforms others, providing the highest quantifiable proteins. Next, we apply three common DIA strategies in characterizing human induced pluripotent stem cell (iPSC)-derived neurons and show single-shot mass spectrometry (MS) using single-CV (-35 V)-FAIMS-DIA results in >9,000 quantifiable proteins with <10% missing values, as well as superior reproducibility and accuracy compared with other existing DIA methods.

iPSC motor neurons, but not other derived cell types, capture gene expression changes in postmortem sporadic ALS motor neurons.

Motor neuron degeneration, the defining feature of amyotrophic lateral sclerosis (ALS), is a primary example of cell-type specificity in neurodegenerative diseases. Using isogenic pairs of induced pluripotent stem cells (iPSCs) harboring different familial ALS mutations, we assess the capacity of iPSC-derived lower motor neurons, sensory neurons, astrocytes, and superficial cortical neurons to capture disease features including transcriptional and splicing dysregulation observed in human postmortem neurons. At early time points, differentially regulated genes in iPSC-derived lower motor neurons, but not other cell types, overlap with one-third of the differentially regulated genes in laser-dissected motor neurons from ALS compared with control postmortem spinal cords. For genes altered in both the iPSC model and bona fide human lower motor neurons, expression changes correlate between the two populations. In iPSC-derived lower motor neurons, but not other derived cell types, we detect the downregulation of genes affected by TDP-43-dependent splicing. This reduction takes place exclusively within genotypes known to involve TDP-43 pathology.

Amino acid homeostasis is a target of metformin therapy.

OBJECTIVE: Unexplained changes in regulation of branched chain amino acids (BCAA) during diabetes therapy with metformin have been known for years. Here we have investigated mechanisms underlying this effect. METHODS: We used cellular approaches, including single gene/protein measurements, as well as systems-level proteomics. Findings were then cross-validated with electronic health records and other data from human material. RESULTS: In cell studies, we observed diminished uptake/incorporation of amino acids following metformin treatment of liver cells and cardiac myocytes. Supplementation of media with amino acids attenuated known effects of the drug, including on glucose production, providing a possible explanation for discrepancies between effective doses in vivo and in vitro observed in most studies. Data-Independent Acquisition proteomics identified that SNAT2, which mediates tertiary control of BCAA uptake, was the most strongly suppressed amino acid transporter in liver cells following metformin treatment. Other transporters were affected to a lesser extent. In humans, metformin attenuated increased risk of left ventricular hypertrophy due to the AA allele of KLF15, which is an inducer of BCAA catabolism. In plasma from a double-blind placebo-controlled trial in nondiabetic heart failure (trial registration: NCT00473876), metformin caused selective accumulation of plasma BCAA and glutamine, consistent with the effects in cells. CONCLUSIONS: Metformin restricts tertiary control of BCAA cellular uptake. We conclude that modulation of amino acid homeostasis contributes to therapeutic actions of the drug.

Mpox Vaccine Interest Survey Prioritization and Data Flow: Maricopa County, Arizona, July-August 2022.

With increasing mpox cases in Maricopa County, Arizona, the county's health department launched a survey on July 11, 2022, to gather eligibility and contact data and provide clinic information to those interested in JYNNEOS as postexposure prophylaxis (PEP) or expanded postexposure prophylaxis(PEP++). Survey data were matched to case and vaccination data. Overall, 343 of the 513 respondents (66.9%) who reported close contact with an mpox case patient received PEP and 1712 of the 3379 respondents (50.7%) who were unsure of their contact status received PEP++. This outreach intervention connected potential close contacts unknown to MCDPH with PEP or PEP++. (Am J Public Health. 2023;113(5):504-508. https://doi.org/10.2105/AJPH.2023.307224).

Plakophilin 2 gene therapy prevents and rescues arrhythmogenic right ventricular cardiomyopathy in a mouse model harboring patient genetics.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a fatal genetic heart disease characterized by cardiac arrhythmias, in which fibrofatty deposition leads to heart failure, with no effective treatments. Plakophilin 2 (PKP2) is the most frequently mutated gene in ARVC, and although altered RNA splicing has been implicated, there are no models to study its effect and therapeutics. Here, we generate a mouse model harboring a PKP2 mutation (IVS10-1G>C) affecting RNA splicing, recapitulating ARVC features and sudden death starting at 4 weeks. Administering AAV-PKP2 gene therapy (adeno-associated viral therapy to drive cardiac expression of PKP2) to neonatal mice restored PKP2 protein levels, completely preventing cardiac desmosomal and pathological deficits associated with ARVC, ensuring 100% survival of mice up to 6 months. Late-stage AAV-PKP2 administration rescued desmosomal protein deficits and reduced pathological deficits including improved cardiac function in adult mice, resulting in 100% survival up to 4 months. We suggest that AAV-PKP2 gene therapy holds promise for circumventing ARVC associated with PKP2 mutations, including splice site mutations.

A reference human induced pluripotent stem cell line for large-scale collaborative studies.

Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between lines makes it challenging to replicate key findings and integrate data across research groups. To address this issue, we sub-cloned candidate human iPSC lines and deeply characterized their genetic properties using whole genome sequencing, their genomic stability upon CRISPR-Cas9-based gene editing, and their phenotypic properties including differentiation to commonly used cell types. These studies identified KOLF2.1J as an all-around well-performing iPSC line. We then shared KOLF2.1J with groups around the world who tested its performance in head-to-head comparisons with their own preferred iPSC lines across a diverse range of differentiation protocols and functional assays. On the strength of these findings, we have made KOLF2.1J and its gene-edited derivative clones readily accessible to promote the standardization required for large-scale collaborative science in the stem cell field.

Effect of chronic administration of 17ß-estradiol on the vasopressor responses induced by the sympathetic nervous system in insulin resistance rats.

Several studies have demonstrated that the underlying mechanism of insulin resistance (IR) is linked with developing diseases like diabetes mellitus, hypertension, metabolic syndrome, and polycystic ovary syndrome. In turn, the dysfunction of female gonadal hormones (especially 17ß-estradiol) may be related to the development of IR complications since different studies have shown that 17ß-estradiol has a cardioprotector and vasorelaxant effect. This study aimed was to determine the effect of the 17ß-estradiol administration in insulin-resistant rats and its effects on cardiovascular responses in pithed rats. Thus, the vasopressor responses are induced by sympathetic stimulation or i.v. bolus injections of noradrenaline (α1/2), methoxamine (α1), and UK 14,304 (α2) adrenergic agonist were determined in female pithed rats with fructose-induced insulin resistance or control rats treated with: 1) 17ß-estradiol or 2) its vehicle (oil) for 5 weeks. Thus, 17ß-estradiol decreased heart rate, prevented the increase of blood pressure induced by ovariectomy, but with the opposite effect on sham-operated rats; and decreased vasopressor responses induced by i.v. bolus injections of noradrenaline on sham-operated (control and fructose group) and ovariectomized (control) rats, and those induced by i.v. bolus injections of methoxamine (α1 adrenergic agonist). Overall, these results suggest 17ß-estradiol has a cardioprotective effect, and its effect on vasopressor responses could be mediated mainly by the α1 adrenergic receptor. In contrast, IR with ovariectomy 17ß-estradiol decreases or loses its cardioprotector effect, this could suggest a possible link between the adrenergic receptors and the insulin pathway.

Prevalencia de afecciones musculoesqueléticas y factores asociados en adultos mayores en una Fundación de Duran / Prevalence of musculoskeletal conditions and associated factors in older adults in a Duran Foundation / Prevalência de condições musculoesqueléticas e fatores associados em idosos de uma Fundação Duran

Las afecciones musculoesqueléticas comprenden trastornos degenerativos que aparecen principalmente en personas de edad avanzada, deteriorando significativamente la salud, debido a que están relacionadas con dolor muscular u óseo, alteraciones del movimiento, mayor riesgo de caídas, fracturas y capacidad alterada para realizar las actividades del diario vivir. Objetivo. Determinar la prevalencia de afecciones musculo esqueléticas y factores asociados en el adulto mayor que asistieron a la Fundación. Metodología. Con enfoque cuantitativo, descriptivo, transversal, no experimental y retrospectivo. Los instrumentos a utilizar incluyeron el dinamómetro, índice de masa corporal y la antropometría. Resultados. se determinó que la prevalencia de las alteraciones musculoesqueléticas fue del 59%, de los cuales, la artrosis de rodilla fue la más común con una prevalencia del 90%, asociados con un índice de masa corporal elevado con un valor mayor a 25 kg/m2 y una fuerza muscular disminuida con un valor menor a 28.2 kg para los hombres y menor a 15.4 kg para las mujeres. En contraste, las medidas antropométricas fueron normales, con valores mayor o igual a 31 cm para la circunferencia de la pantorrilla y mayor o igual a 22 cm para la circunferencia del brazo. Conclusión. Existe una alta prevalencia de alteraciones musculoesqueléticas, El sobrepeso y la debilidad muscular fueron los únicos factores asociados.

Musculoskeletal disorders include degenerative disorders that appear mainly in elderly people, significantly deteriorating their health, because they are related to muscle or bone pain, movement disorders, increased risk of falls, fractures and impaired ability to perform daily activities. Objective. To determine the prevalence of musculoskeletal disorders and associated factors in older adults attending the Fundación. Methodology. With a quantitative, descriptive, cross-sectional, non-experimental and retrospective approach. The instruments to be used included the dynamometer, body mass index and anthropometry. Results. It was determined that the prevalence of musculoskeletal alterations was 59%, of which knee osteoarthritis was the most common with a prevalence of 90%, associated with an elevated body mass index with a value greater than 25 kg/m2 and decreased muscle strength with a value of less than 28.2 kg for men and less than 15.4 kg for women. In contrast, anthropometric measurements were normal, with values greater than or equal to 31 cm for calf circumference and greater than or equal to 22 cm for arm circumference. Conclusion. There is a high prevalence of musculoskeletal alterations. Overweight and muscle weakness were the only associated factors

As condições musculoesqueléticas compreendem desordens degenerativas que ocorrem principalmente em pessoas idosas, prejudicando significativamente a saúde, pois estão associadas a dores musculares ou nos ossos, comprometimento dos movimentos, aumento do risco de quedas, fraturas e incapacidade de realizar atividades da vida diária. Objetivo. Para determinar a prevalência de distúrbios musculoesqueléticos e fatores associados em adultos idosos que frequentam a Fundação. Metodologia. Com uma abordagem quantitativa, descritiva, transversal, não-experimental e retrospectiva. Os instrumentos utilizados incluíam o dinamômetro, o índice de massa corporal e a antropometria. Resultados. Foi determinado que a prevalência de distúrbios musculoesqueléticos foi de 59%, dos quais a osteoartrose do joelho foi a mais comum com uma prevalência de 90%, associada a um alto índice de massa corporal com um valor superior a 25 kg/m2 e uma diminuição da força muscular com um valor inferior a 28,2 kg para homens e inferior a 15,4 kg para mulheres. Em contraste, as medidas antropométricas foram normais, com valores maiores ou iguais a 31 cm para a circunferência da barriga da perna e maiores ou iguais a 22 cm para a circunferência do braço. Conclusão. Há uma alta prevalência de distúrbios musculoesqueléticos. O excesso de peso e a fraqueza muscular foram os únicos fatores associados.