A Pillar-Based High-Throughput Myogenic Differentiation Assay to Assess Drug Safety

High-throughput, pillar-strip-based assays have been proposed as a drug-safety screening tool for developmental toxicity. In the assay described here, muscle cell culture and differentiation were allowed to occur at the end of a pillar strip (eight pillars) compatible with commercially available 96-well plates. Previous approaches to characterize cellular differentiation with immunostaining required a burdensome number of washing steps; these multiple washes also resulted in a high proportion of cellular loss resulting in poor yield. To overcome these limitations, the approach described here utilizes cell growth by easily moving the pillars for washing and immunostaining without significant loss of cells. Thus, the present pillar-strip approach is deemed suitable for monitoring high-throughput myogenic differentiation. Using this experimental high-throughput approach, eight drugs (including two well-known myogenic inhibitory drugs) were tested at six doses in triplicate, which allows for the generation of dose–response curves of nuclei and myotubes in a 96-well platform. As a result of comparing these F-actin (an actin-cytoskeleton protein), nucleus, and myotube data, two proposed differentiation indices—curve-area-based differentiation index (CA-DI) and maximum-point-based differentiation index (MP-DI) were generated. Both indices successfully allowed for screening of high-myogenic inhibitory drugs, and the maximum-point-based differentiation index (MP-DI) experimentally demonstrated sensitivity for quantifying drugs that inhibited myogenic differentiation.

SSR Marker-Based Genetic Resource Assessment of The Rainbow Clam Moerella Iridescens Along The Coasts of China: Implications for Strategy of Conservation Management

This study aims to determine the genetic structure and diversity of rainbow clam Moerella iridescens in different sea areas of China. Seventeen pairs of microsatellite primers (SSR) were used to amplify the SSRs of rainbow clam in Lianyungang of Haizhou Bay, Chongming of Shanghai, Ningde in Fujian, Cixi and Wenzhou in Zhejiang. A total of 1146 alleles were detected in 310 individuals from the 17 SSR loci. The average observed heterozygosity of six populations was 0.4381–0.6139, the average expected heterozygosity was 0.5897–0.7325, and the average Shannon diversity index was 1.2655–1.7998. The clams exhibited rich genetic diversity and the FST of the genetic differentiation index of the six populations was 0.0470, indicating low genetic differentiation amongst the populations. The results indictated that rainbow clam along China coasts exhibited high diversity and low population differentiation.

Geographical Structuring of Quercus robur (L.) Chloroplast DNA Haplotypes in Lithuania: Recolonization, Adaptation, or Overexploitation Effects?

We studied the maternally inherited chloroplast DNA polymorphism at three microsatellite loci of 157 Quercus robur trees from 38 native populations in Lithuania. We found high diversity of eight haplotypes from the Balkan lineage A (frequency 0.75) and the “German” subbranch of the Balkan lineage A (freq. 0.12), western and eastern Italian lineages C (freq. 0.05 and 0.06, respectively), and Iberian lineage B (freq. 0.03). The haplotypes were geographically well structured (among population differentiation index PhiPT = 0.30, the p-value < 0.001) that is unexpected for such a small territory as Lithuania. We raised a hypothesis on historical overexploitation of oaks by eliminating certain haplotypes in Lithuania, following a drastic felling of oak forests over the last few centuries.

Immune response of the harderian gland in chickens to infectious bronchitis coronavirus

Aim. To determine the difference in immune responses of the harderian gland in clinically healthy chickens and the ones with infectious bronchitis based on the content, localization and morphometric estimation of the surface markers of Т- and В-lymphocytes and to determine the differentiation index as an indicator of assessing body defenses. Methods. Histological, immunohistochemical, optical, morphometric and statistical. Results. The histological study of the harderian gland of chickens with infectious bronchitis determined the swelling and proliferation of the connective tissue as well as infiltration of secretory lobules by lymphoid cells. It was found that the immunity of chickens with infectious bronchitis, in which the harderian gland plays a relevant role, depends considerably on the differentiation index of immunocompetent cells. There was a reliable 1.77- and 1.36-fold decrease in this indicator for 40- and 90-day-old chickens, respectively, in case of nephroso-nephritic form of infectious bronchitis which demonstrated a weaker function of the defense cells of this organ. According to the cytomorphometric analysis, the number of cells, expressing CD4+, CD8+, CD20+, CD45RA+ markers in the harderian gland of sick 20-, 40-, and 90-day-old chickens with respiratory and nephroso-nephritic forms of infectious bronchitis was reliably (P < 0.05) increasing compared to the clinically healthy chickens. For instance, the number of mature В-lymphocytes increased in sick 20-day-old chickens – 2.44 times, 40-day-old chickens – 1.88 times, and 90-day-old ones – 2.62 times compared to clinically healthy chickens. Conclusions. The data were obtained about the changes in quantitative and qualitative composition of lymphocytes with surface markers CD4+, CD8+, CD20+, CD45RA+ in the harderian gland of chickens with infectious bronchitis. Our results will supplement current knowledge about the feasibility of immunohistochemical methods in the diagnostics of avian infectious bronchitis.

Tree Species Diversity and Spatial Distribution of Aleppo Pine Stands in Northeastern Algeria

The spatial pattern of species is one of the key studied parameters in ecology so as to better understand the ecological processes and the functioning of forest ecosystems. This paper describes the classification of structural indices measuring the alpha diversity and examines typical representatives of the classification groups such as the Shannon’s index, aggregation index by Clark and Evans, the mingling index, the diameter differentiation index and the coefficient of segregation by Pielou. The tree inventory made it possible to count 7 species that are divided into six (06) families. Only Pinus halepensis Mill. trees were taken into account via calculation in spatial distribution. Western exposure shows the most regular tree patterns (1.6±0.1) according to the aggregation index by Clark and Evans, while the species mingling index for south- and east-facing stands indicates segregation of Pinus halepensis Mill. groups. The diameter differentiation index for the majority of the studied stands is assumed through estimated values within the range that starts from 0.4 to 0.9 for the four exposures. The distribution shows that western and eastern exposures belong to the fourth class of differentiation (very large differentiation), which means that the trees with the smallest DBH have less than 30% of the size of the neighbouring trees since the diameter differentiation index for the two exposures is 0.9±0.05 and 0.7±0.2.

High-quality carnivoran genomes from roadkill samples enable comparative species delineation in aardwolf and bat-eared fox

In a context of ongoing biodiversity erosion, obtaining genomic resources from wildlife is essential for conservation. The thousands of yearly mammalian roadkill provide a useful source material for genomic surveys. To illustrate the potential of this underexploited resource, we used roadkill samples to study the genomic diversity of the bat-eared fox (Otocyon megalotis) and the aardwolf (Proteles cristatus), both having subspecies with similar disjunct distributions in Eastern and Southern Africa. First, we obtained reference genomes with high contiguity and gene completeness by combining Nanopore long reads and Illumina short reads. Then, we showed that the two subspecies of aardwolf might warrant species status (P. cristatus and P. septentrionalis) by comparing their genome-wide genetic differentiation to pairs of well-defined species across Carnivora with a new Genetic Differentiation index (GDi) based on only a few resequenced individuals. Finally, we obtained a genome-scale Carnivora phylogeny including the new aardwolf species.

Momentary Emotion Differentiation: The Derivation and Validation of an index to Study Within-Person Fluctuations in Emotion Differentiation

Emotion differentiation refers to the tendency to label emotions in a granular way. While differentiation is an important individual difference in the context of psychological well-being, it is unknown how it fluctuates within individuals. Such a within-person measure is important, since it would allow the study of how changes in differentiation predict subsequent levels of other variables of interest. Here, we present a framework to study emotion differentiation at the within-person level by introducing a momentary emotion differentiation index. This index is directly derived from the classical emotion differentiation index, the intraclass correlation. We first give a theoretical derivation of this index. Next, using data from two experience sampling studies, we show how this new momentary index is related to other momentary indicators of well-being, and take the first steps in building its nomological network. A better understanding of within-person fluctuations in emotion differentiation will allow us to identify the causes and consequences of these fluctuations, and search for ways to teach individuals to increase their level of emotion differentiation.

Momentary Emotion Differentiation: The derivation and validation of a framework to study within-person fluctuations in emotion differentiation

Emotion differentiation refers to the tendency to label emotions in a granular way. While differentiation is an important individual difference in the context of psychological well-being (Kashdan et al., 2015), it is unknown how it fluctuates within individuals. Such a within-person measure is important, since it would allow the study of how changes in differentiation predict subsequent levels of other variables of interest. Here, we present a framework to study emotion differentiation at the within-person level by introducing a momentary emotion differentiation index. This index is directly derived from the classical emotion differentiation index, the intra-class correlation. We first give a theoretical derivation of this index. Next, using data from two experience sampling studies, we show how this new momentary index is related to other momentary indicators of well-being, and take the first steps in building its nomological network. A better understanding of within-person fluctuations in emotion differentiation will allow us to identify the causes and consequences of these fluctuations, and search for ways to teach individuals to increase their level of emotion differentiation.