A TALEN Genome-Editing System for Generating Human Stem Cell-Based Disease Models

SummaryTranscription activator-like effector nucleases (TALENs) are a new class of engineered nucleases that are easier to design to cleave at desired sites in a genome than previous types of nucleases. We report here the use of TALENs to rapidly and efficiently generate mutant alleles of 15 genes in cultured somatic cells or human pluripotent stem cells, the latter for which we differentiated both the targeted lines and isogenic control lines into various metabolic cell types. We demonstrate cell-autonomous phenotypes directly linked to disease—dyslipidemia, insulin resistance, hypoglycemia, lipodystrophy, motor-neuron death, and hepatitis C infection. We found little evidence of TALEN off-target effects, but each clonal line nevertheless harbors a significant number of unique mutations. Given the speed and ease with which we were able to derive and characterize these cell lines, we anticipate TALEN-mediated genome editing of human cells becoming a mainstay for the investigation of human biology and disease

Prevalence of Dementia and Mild Cognitive Impairment in Indigenous Bolivian Forager-Horticulturalists

Introduction We evaluated the prevalence of dementia and mild cognitive impairment (MCI) in indigenous Tsimane and Moseten, who lead a subsistence lifestyle. Methods Participants from population-based samples ≥ 60 years of age (n = 623) were assessed using adapted versions of the Modified Mini-Mental State Examination, informant interview, longitudinal cognitive testing and brain computed tomography (CT) scans. Results Tsimane exhibited five cases of dementia (among n = 435; crude prevalence = 1.2%, 95% confidence interval [CI]: 0.4, 2.7); Moseten exhibited one case (among n = 169; crude prevalence = 0.6%, 95% CI: 0.0, 3.2), all age ≥ 80 years. Age-standardized MCI prevalence was 7.7% (95% CI: 5.2, 10.3) in Tsimane and 9.8% (95% CI: 4.9, 14.6) in Moseten. Cognitive impairment was associated with visuospatial impairments, parkinsonian symptoms, and vascular calcification in the basal ganglia. Discussion The prevalence of dementia in this cohort is among the lowest in the world. Widespread intracranial medial arterial calcifications suggest a previously unrecognized, non-Alzheimer\u27s disease (AD) dementia phenotype

A TALEN Genome-Editing System for Generating Human Stem Cell-Based Disease Models

Transcription activator-like effector nucleases (TALENs) are a new class of engineered nucleases that are easier to design to cleave at desired sites in a genome than previous types of nucleases. We report here the use of TALENs to rapidly and efficiently generate mutant alleles of 15 genes in cultured somatic cells or human pluripotent stem cells, the latter for which we differentiated both the targeted lines and isogenic control lines into various metabolic cell types. We demonstrate cell-autonomous phenotypes directly linked to disease—dyslipidemia, insulin resistance, hypoglycemia, lipodystrophy, motor-neuron death, and hepatitis C infection. We found little evidence of TALEN off-target effects, but each clonal line nevertheless harbors a significant number of unique mutations. Given the speed and ease with which we were able to derive and characterize these cell lines, we anticipate TALEN-mediated genome editing of human cells becoming a mainstay for the investigation of human biology and disease.Stem Cell and Regenerative Biolog

T cell phenotypes in COVID-19 - a living review

COVID-19 is characterized by profound lymphopenia in the peripheral blood, and the remaining T cells display altered phenotypes, characterized by a spectrum of activation and exhaustion. However, antigen-specific T cell responses are emerging as a crucial mechanism for both clearance of the virus and as the most likely route to long-lasting immune memory that would protect against re-infection. Therefore, T cell responses are also of considerable interest in vaccine development. Furthermore, persistent alterations in T cell subset composition and function post-infection have important implications for patients’ long-term immune function. In this review, we examine T cell phenotypes, including those of innate T cells, in both peripheral blood and lungs, and consider how key markers of activation and exhaustion correlate with, and may be able to predict, disease severity. We focus on SARS-CoV-2-specific T cells to elucidate markers that may indicate formation of antigen-specific T cell memory. We also examine peripheral T cell phenotypes in recovery and the likelihood of long-lasting immune disruption. Finally, we discuss T cell phenotypes in the lung as important drivers of both virus clearance and tissue damage. As our knowledge of the adaptive immune response to COVID-19 rapidly evolves, it has become clear that while some areas of the T cell response have been investigated in some detail, others, such as the T cell response in children remain largely unexplored. Therefore, this review will also highlight areas where T cell phenotypes require urgent characterisation

The role and uses of antibodies in COVID-19 infections: a living review

Coronavirus disease 2019 has generated a rapidly evolving field of research, with the global scientific community striving for solutions to the current pandemic. Characterizing humoral responses towards SARS-CoV-2, as well as closely related strains, will help determine whether antibodies are central to infection control, and aid the design of therapeutics and vaccine candidates. This review outlines the major aspects of SARS-CoV-2-specific antibody research to date, with a focus on the various prophylactic and therapeutic uses of antibodies to alleviate disease in addition to the potential of cross-reactive therapies and the implications of long-term immunity

MARK chick survival models

Zipped file contains two files: 1) a MARK DBF file, and 2) a MARK FTP file. Both are required to open and view MARK chick survival models. To view, unzip files into the same folder. Requires Program MARK (www.phidot.org) to open

Data from: Mismatches between breeding phenology and resource abundance of resident alpine ptarmigan negatively affect chick survival

1. Phenological mismatches – defined here as the difference in reproductive timing of an individual relative to the availability of its food resources – occur in many avian species. Mistiming breeding activities in environments with constrained breeding windows may have severe fitness costs due to reduced opportunities for repeated breeding attempts. Therefore, species occurring in alpine environments may be particularly vulnerable. 2. We studied fitness consequences of timing of breeding in an alpine-endemic species, the white-tailed ptarmigan (Lagopus leucura), to investigate its influence on chick survival. We estimated phenological mismatch by measuring plant and arthropods used by ptarmigan in relation to their timing of breeding. 3. We monitored 120 nests and 67 broods over a three-year period (2013–2015) at three alpine study sites in the Rocky Mountains of Colorado. During this same period we actively monitored food resource abundance in brood-use areas to develop year and site specific resource phenology curves. We developed several mismatch indices from these curves that were then fit as covariates in mark-recapture chick survival models. 4. A correlation analysis between seasonal changes in arthropod and food plant abundance indicated that a normalized difference vegetation index (NDVI) was likely the best predictor for food available to hens and chicks. A survival model that included an interaction between NDVI mismatch and chick age received strong support and indicated young chicks were more susceptible to mismatch than older chicks. 5. We provide evidence that individual females of a resident alpine species can be negatively affected by phenological mismatch. Our study focused on individual females and did not examine if phenological mismatch was present at the population level. Future work in animal populations occurring in mountain systems focusing on a combination of both individual- and population-level metrics of mismatch will be beneficial

Estimates of Southern White-tailed Ptarmigan daily nest survival from multiple sites in the Southern Rocky Mountains of Colorado

Estimating vital rates of avian species is important to understand population dynamics and develop potential conservation strategies that target rates for management. Avian species have reduced potential for high annual fecundity in alpine ecosystems due to a short breeding window and harsh weather conditions. We located nests from Southern White-tailed Ptarmigan ( Lagopus leucura altipetens ) across six study sites in the Southern Rocky Mountains of Colorado to estimate daily nest survival from 2013–2017. We used a known-fate hierarchical nest survival model and fit several covariates, including environmental conditions representing daily weather events and shrub cover, to describe variation in daily survival and derive estimates of nest success. We located and monitored 198 nests from 128 radio-marked ptarmigan hens. The mean nest success estimated as a derived parameter from daily nest survival was 45.6% (95% credible interval [CI]: 31.2–59.6%) and ranged from 40.3% to 50.3% across sites. Variation in daily nest survival was poorly described by the covariates we fit (95% CI of most slope coefficients overlapped 0), although there was some support for a negative effect of relative elevation (nests at lower elevations within a site survived at higher rates) and a positive effect of nest age (older nests survived at higher rates). We examined how variation in nest success was likely to influence the finite rate of population growth using a simple simulation with an age-transition matrix parameterized with previously reported fecundity and survival estimates. We found that the finite growth rate was predicted to increase 18.7% when evaluated from the lower to upper 95% CI estimated values of nest success, conditional on the other vital rates used in our simulation. We discuss the broader implications of these findings in the context of managing for nest survival of Southern White-tailed Ptarmigan