Molecular Reclassification of Crohn’s Disease: A Cautionary Note on Population Stratification

Complex human diseases commonly differ in their phenotypic characteristics, e.g., Crohn’s disease (CD) patients are heterogeneous with regard to disease location and disease extent. The genetic susceptibility to Crohn’s disease is widely acknowledged and has been demonstrated by identification of over 100 CD associated genetic loci. However, relating CD subphenotypes to disease susceptible loci has proven to be a difficult task. In this paper we discuss the use of cluster analysis on genetic markers to identify genetic-based subgroups while taking into account possible confounding by population stratification. We show that it is highly relevant to consider the confounding nature of population stratification in order to avoid that detected clusters are strongly related to population groups instead of disease-specific groups. Therefore, we explain the use of principal components to correct for population stratification while clustering affected individuals into genetic-based subgroups. The principal components are obtained using 30 ancestry informative markers (AIM), and the first two PCs are determined to discriminate between continental origins of the affected individuals. Genotypes on 51 CD associated single nucleotide polymorphisms (SNPs) are used to perform latent class analysis, hierarchical and Partitioning Around Medoids (PAM) cluster analysis within a sample of affected individuals with and without the use of principal components to adjust for population stratification. It is seen that without correction for population stratification clusters seem to be influenced by population stratification while with correction clusters are unrelated to continental origin of individuals.     

Evolution of pseudogenes in the immunoglobulin V H-gene family of the mouse

A quantitative analysis of the complexity of the J558 V _H -gene family in the mouse immunoglobulin heavy chain (Igh) gene locus has been performed. Considerable variations in the degree of complexity are observed in various Igh haplotypes derived from laboratory mice and wild mice. The BALB/c strain shows the highest degree of complexity of the J558 V _H -gene family when all mice are compared. Multiple gene duplications seem to have occurred in the BALB/c-derived J558 V_H-gene family less than 1–2 million years ago. This dating is supported by the divergence in coding and flanking regions of three strongly homologous V _H -region genes. Two of these genes were generated by the duplication of a pseudogene about 1.5 × 10⁵ years ago. A recent expansion of the J558 V _H -gene family and therefore little time for evolutionary drift may explain why most of the pseudogenes in this family exhibit a largely intact structure. We also describe two V _H -region genes which represent older pseudogenes in states of progressive disintegration.     

Phylogenetic distribution in the genus Mus of t-complex-specific DNA and protein markers: inferences on the origin of t-haplotypes

We have examined the phylogenetic distribution of two t-specific markers among representatives of various taxa belonging to the genus Mus. The centromeric TCP-1a marker (a testicular protein variant specific for all t-haplotypes so far studied) has also been apparently detected in several non-t representatives of the Mus IVA, Mus IVB, and probably M. cervicolor species. By contrast, a t-specific restriction- fragment-length polymorphism allele (RFLP) of the telomeric alpha- globin pseudogene DNA marker alpha-psi-4 was found only in animals belonging to the M. musculus-complex species either bearing genuine t- haplotypes or, like the M. m. bactrianus specimen studied here, likely to do so. This t-specific alpha-psi-4 RFLP allele was found to be as divergent from the RFLP alleles of the latter, non-t, taxonomical groups as it is from Mus 4A, Mus 4B, or M. spretus ones. These results suggest the presence of t-haplotypes and of t-specific markers in populations other than those belonging to the M. m. domesticus and M. m. musculus subspecies, implying a possible origin for t-haplotypes prior to the radiation of the most recent offshoot of the Mus genus (i.e., the spretus/domesticus divergence), some 1-3 Myr ago.      

A theoretical analysis of radiation interception in a two-species plant canopy.

Classical radiation interception laws for monospecific canopies cannot be used directly for bispecific canopies. They are always based on the gap frequency concept (i.e., the probability of no interception), which does not provide any information about the sharing of intercepted radiation between species. A theoretical analysis is reported that relates the radiation interception probabilities to the geometrical structure of the crop (i.e., the leaf area density and the leaf angle distribution of each component) and the foliage dispersion. The leaf dispersion globally describes the spatial relations between the leaf elements; it may be regular if the leaves avoid mutual shading, random, or clumped if they tend to overlap. For such two-species canopies, the leaf dispersions within each component (WSLD: within-species leaf dispersion) and between two species (BSLD: between-species leaf dispersion) are distinguished. Using bivariate multinomial distributions, general expressions for the gap frequency and the interception probabilities of a homogeneous vegetation layer were set as exponential functions of the foliage thickness, taking into account a number of dispersion parameters as small as possible. First, one WSLD for each species describes the rate of foliage overlap between the leaves of this species; it is quite similar to the leaf dispersion of single-species canopies. Second, the rate of foliage overlap between species is characterized by one BSLD. As in monospecific canopies, this parameter is positive, zero, or negative, respectively, for regular, random, or clumped BSLD. Third, another BSLD parameter has to be used if the foliage overlap between species is more than random (i.e., in the case of clumped BSLD); the latter shows the direction of overlap between species and may be taken as the probability of finding a leaf element of the first species in the case of marked overlapping. Suggestions for estimating the leaf dispersion parameters and possible uses of such relations are also discussed.     

H-2 Polymorphisms Are More Uniformly Distributed than Allozyme Polymorphisms in Natural Populations of House Mice

Patterns of H-2 and allozyme polymorphism in natural populations of house mice from Europe, North Africa and South America were analyzed. The purpose of the analysis was to determine whether H-2 and allozyme polymorphisms were similarly distributed both geographically and temporally in wild mice. Two subspecies of house mice, Mus musculus domesticus and M. m. musculus were sampled and the polymorphisms of two H-2 class I genes, H-2K and H-2D, and 34 allozyme-encoding genes were surveyed. The three kinds of analyses that were conducted included a hierarchical gene diversity analysis, an analysis of the effects of barriers to gene flow, and an analysis of similarity networks. Each of the comparisons demonstrated that H-2 polymorphisms were more uniformly distributed than allozyme polymorphisms and provided additional evidence that H-2 and allozyme polymorphisms are subject to different evolutionary pressures. The analysis of similarity networks also demonstrated that H-2 genes provide little information about the phylogeny of wild mice.     

The separation of the α-chains of collagen by free-flow electrophoresis

1. The separation of the alpha-components of codfish-skin and calf-skin collagen from the higher-molecular-weight components was accomplished by gel filtration with Bio-Gel P-300 resin in 5m-lithium chloride and at pH7.4. 2. From this fraction three distinct alpha-components, corresponding to the alpha1-, alpha2- and alpha3- chains of collagen, were isolated by free-flow electrophoresis in 6m-urea at 4 degrees . At the temperatures and pH values used there was no detectable degradation of the alpha-chains.     

Increase in nuclear aluminum concentration during hepatic regeneration: study by analytical ionic microscopy

20 hrs. after partial hepatectomy, a significant increase in the aluminum concentration of liver cell nuclei has been measured by analytical ion microscopy. These results indicate a possible role of aluminium in an in vivo induced proliferative response.     

The distribution of coastal fish eDNA sequences in the Anthropocene

Aim

Coastal fishes have a fundamental role in marine ecosystem functioning and contributions to people, but face increasing threats due to climate change, habitat degradation and overexploitation. The extent to which human pressures are impacting coastal fish biodiversity in comparison with geographic and environmental factors at large spatial scale is still under scrutiny. Here, we took advantage of environmental DNA (eDNA) metabarcoding to investigate the relationship between fish biodiversity, including taxonomic and genetic components, and environmental but also socio-economic factors.

Location

Tropical, temperate and polar coastal areas.

Time period

Present day.

Major taxa studied

Marine fishes.

Methods

We analysed fish eDNA in 263 stations (samples) in 68 sites distributed across polar, temperate and tropical regions. We modelled the effect of environmental, geographic and socio-economic factors on α- and β-diversity. We then computed the partial effect of each factor on several fish biodiversity components using taxonomic molecular units (MOTU) and genetic sequences. We also investigated the relationship between fish genetic α- and β-diversity measured from our barcodes, and phylogenetic but also functional diversity.

Results

We show that fish eDNA MOTU and sequence α- and β-diversity have the strongest correlation with environmental factors on coastal ecosystems worldwide. However, our models also reveal a negative correlation between biodiversity and human dependence on marine ecosystems. In areas with high dependence, diversity of all fish, cryptobenthic fish and large fish MOTUs declined steeply. Finally, we show that a sequence diversity index, accounting for genetic distance between pairs of MOTUs, within and between communities, is a reliable proxy of phylogenetic and functional diversity.

Main conclusions

Together, our results demonstrate that short eDNA sequences can be used to assess climate and direct human impacts on marine biodiversity at large scale in the Anthropocene and can further be extended to investigate biodiversity in its phylogenetic and functional dimensions.     

A major difference between the divergence patterns within the lines-1 families in mice and voles

L1 retroposons are represented in mice by subfamilies of interspersed sequences of varied abundance. Previous analyses have indicated that subfamilies are generated by duplicative transposition of a small number of members of the L1 family, the progeny of which then become a major component of the murine L1 population, and are not due to any active processes generating homology within preexisting groups of elements in a particular species. In mice, more than a third of the L1 elements belong to a clade that became active approximately 5 Mya and whose elements are > or = 95% identical. We have collected sequence information from 13 L1 elements isolated from two species of voles (Rodentia: Microtinae: Microtus and Arvicola) and have found that divergence within the vole L1 population is quite different from that in mice, in that there is no abundant subfamily of homologous elements. Individual L1 elements from voles are very divergent from one another and belong to a clade that began a period of elevated duplicative transposition approximately 13 Mya. Sequence analyses of portions of these divergent L1 elements (approximately 250 bp each) gave no evidence for concerted evolution having acted on the vole L1 elements since the split of the two vole lineages approximately 3.5 Mya; that is, the observed interspecific divergence (6.7%-24.7%) is not larger than the intraspecific divergence (7.9%-27.2%), and phylogenetic analyses showed no clustering into Arvicola and Microtus clades.