Genetic confounding of the relationship between father absence and age at menarche

Research in evolutionary psychology, and life history theory in particular, has yielded important insights into the developmental processes that underpin variation in growth, psychological functioning, and behavioral outcomes across individuals. Yet, there are methodological concerns that limit the ability to draw causal inferences about human development and psychological functioning within a life history framework. The current study used a simulation-based modeling approach to estimate the degree of genetic confounding in tests of a well-researched life history hypothesis: that father absence (X) is associated with earlier age at menarche (Y). The results demonstrate that the genetic correlation between X and Y can confound the phenotypic association between the two variables, even if the genetic correlation is small—suggesting that failure to control for the genetic correlation between X and Y could produce a spurious phenotypic correlation. We discuss the implications of these results for research on human life history, and highlight the utility of incorporating genetically sensitive tests into future life history research.     

Marked seasonal variation in the wild mouse gut microbiota

Recent studies have provided an unprecedented view of the microbial communities colonizing captive mice; yet the host and environmental factors that shape the rodent gut microbiota in their natural habitat remain largely unexplored. Here, we present results from a 2-year 16 S ribosomal RNA gene sequencing-based survey of wild wood mice (Apodemus sylvaticus) in two nearby woodlands. Similar to other mammals, wild mice were colonized by 10 bacterial phyla and dominated by the Firmicutes, Bacteroidetes and Proteobacteria. Within the Firmicutes, the Lactobacillus genus was most abundant. Putative bacterial pathogens were widespread and often abundant members of the wild mouse gut microbiota. Among a suite of extrinsic (environmental) and intrinsic (host-related) factors examined, seasonal changes dominated in driving qualitative and quantitative differences in the gut microbiota. In both years examined, we observed a strong seasonal shift in gut microbial community structure, potentially due to the transition from an insect- to a seed-based diet. This involved decreased levels of Lactobacillus, and increased levels of Alistipes (Bacteroidetes phylum) and Helicobacter. We also detected more subtle but statistically significant associations between the gut microbiota and biogeography, sex, reproductive status and co-colonization with enteric nematodes. These results suggest that environmental factors have a major role in shaping temporal variations in microbial community structure within natural populations.     

Comparison of marker types and map assumptions using Markov chain Monte Carlo-based linkage analysis of COGA data

We performed multipoint linkage analysis of the electrophysiological trait ECB21 on chromosome 4 in the full pedigrees provided by the Collaborative Study on the Genetics of Alcoholism (COGA). Three Markov chain Monte Carlo (MCMC)-based approaches were applied to the provided and re-estimated genetic maps and to five different marker panels consisting of microsatellite (STRP) and/or SNP markers at various densities. We found evidence of linkage near the GABRB1 STRP using all methods, maps, and marker panels. Difficulties encountered with SNP panels included convergence problems and demanding computations.     

Cleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cells

Linking the heavy chain (HC) and light chain (LC) genes required for monoclonal antibodies (mAb) production on a single cassette using 2A peptides allows control of LC and HC ratio and reduces non-expressing cells. Four 2A peptides derived from the foot-and-mouth disease virus (F2A), equine rhinitis A virus (E2A), porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A), respectively, were compared for expression of 3 biosimilar IgG1 mAbs in Chinese hamster ovary (CHO) cell lines. HC and LC were linked by different 2A peptides both in the absence and presence of GSG linkers. Insertion of a furin recognition site upstream of 2A allowed removal of 2A residues that would otherwise be attached to the HC. Different 2A peptides exhibited different cleavage efficiencies that correlated to the mAb expression level. The relative cleavage efficiency of each 2A peptide remains similar for expression of different IgG1 mAbs in different CHO cells. While complete cleavage was not observed for any of the 2A peptides, GSG linkers did enhance the cleavage efficiency and thus the mAb expression level. T2A with the GSG linker (GT2A) exhibited the highest cleavage efficiency and mAb expression level. Stably amplified CHO DG44 pools generated using GT2A had titers 357, 416 and 600 mg/L for the 3 mAbs in shake flask batch cultures. Incomplete cleavage likely resulted in incorrectly processed mAb species and aggregates, which were removed with a chromatin-directed clarification method and protein A purification. The vector and methods presented provide an easy process beneficial for both mAb development and manufacturing.     

An organ culture of adult mouse skin: An in vitro model for studying the molecular mechanism of skin tumor promotion

A simple method to culture explants of adult mouse skin in a modified Eagle's HeLa cell medium was developed in order to further study the biochemical responses to the tumor promoting phorbol esters. The skin explants remained viable for at least 48 hr, as determined by their ability to incorporate ³H-thymidine into DNA as well as to induce epidermal ornithine decarboxylase (EC 4.1.1.17) activity following 12-0-tetradecanoylphorbol-13-acetate addition. The time course of induction of ornithine decarboxylase activity by the tumor promoter was similar to that observed with intact mice. Furthermore, the addition of retinoic acid and indomethacin, the agents that are known to inhibit the induction of ornithine decarboxylase activity by topically applied TPA, also inhibited the induction of ornithine decarboxylase activity by TPA in skin explants.     

Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system.     

The Cytoplasmic pH Influences Hyphal Tip Growth and Cytoskeleton-Related Organization

The regulatory role of protons in hyphal tip growth was investigated by using membrane-permeant weak acids to acidify cytoplasm of the oomycete Saprolegnia ferax. Acetic acid decreased cytoplasmic pH from approximately pH 7.2 to 6.8, as shown by SNARF-1 measurements of cytoplasmic pH. Inhibition of growth in a dose-dependent manner by acetic, propionic, and isobutyric acid was accompanied by changes in positioning and morphology of mitochondria and nuclei, condensation of chromatin, disruptions in peripheral actin, and increases in hyphal diameter. These cellular alterations were fully reversible, and during recovery, major cytoplasmic movements and extensive apical vacuolations were observed. The results are consistent with proton regulation of the cytoskeleton, nuclear matrix, and/or chromosomes. However, a macroscopic cytoplasmic gradient of H+ in hyphae was not revealed by SNARF-1, indicating that if such a H+ gradient were required, it must occur at a finer level than we detected.     

Absence of phosphorylation of retinoid-binding proteins by protein kinase C in vitro

Cellular retinol-binding protein, cellular retinoic acid-binding protein, and fetal cellular retinol-binding protein were purified to homogeneity and each polypeptide had a molecular weight of 16,000. Their apoproteins were not phosphorylated under the same conditions. Their holoproteins did not inhibit the phosphorylation of histone III-S by protein kinase C. Each of these observations is contrary to the results reported by Cope et al. (Biochem. Biophys. Res. Commun., 120, 593-601, 1984).     

Landscape genetics of an endangered lemur (Propithecus tattersalli) within its entire fragmented range

Habitat fragmentation may strongly reduce individuals’ dispersal among resource patches and hence influence population distribution and persistence. We studied the impact of landscape heterogeneity on the dispersal of the golden‐crowned sifaka (Propithecus tattersalli), an endangered social lemur species living in a restricted and highly fragmented landscape. We combined spatial analysis and population genetics methods to describe population units and identify the environmental factors which best predict the rates and patterns of genetic differentiation within and between populations. We used non‐invasive methods to genotype 230 individuals at 13 microsatellites in all the main forest fragments of its entire distribution area. Our analyses suggest that the Manankolana River and geographical distance are the primary structuring factors, while a national road crossing the region does not seem to impede gene flow. Altogether, our results are in agreement with a limited influence of forest habitat connectivity on gene flow patterns (except for North of the species’ range), suggesting that dispersal is still possible today among most forest patches for this species. Within forest patches, we find that dispersal is mainly among neighbouring social groups, hence confirming previous behavioural observations.