Heterozygosity at neutral and immune loci is not associated with neonatal mortality due to microbial infection in Antarctic fur seals

Numerous studies have reported correlations between the heterozygosity of genetic markers and fitness. These heterozygosity–fitness correlations (HFCs) play a central role in evolutionary and conservation biology, yet their mechanistic basis remains open to debate. For example, fitness associations have been widely reported at both neutral and functional loci, yet few studies have directly compared the two, making it difficult to gauge the relative contributions of genome‐wide inbreeding and specific functional genes to fitness. Here, we compared the effects of neutral and immune gene heterozygosity on death from bacterial infection in Antarctic fur seal (Arctocephalus gazella) pups. We specifically developed a panel of 13 microsatellites from expressed immune genes and genotyped these together with 48 neutral loci in 234 individuals, comprising 39 pups that were classified at necropsy as having most likely died of bacterial infection together with a five times larger matched sample of healthy surviving pups. Identity disequilibrium quantified from the neutral markers was positive and significant, indicative of variance in inbreeding within the study population. However, multilocus heterozygosity did not differ significantly between healthy and infected pups at either class of marker, and little evidence was found for fitness associations at individual loci. These results support a previous study of Antarctic fur seals that found no effects of heterozygosity at nine neutral microsatellites on neonatal survival and thereby help to refine our understanding of how HFCs vary across the life cycle. Given that nonsignificant HFCs are underreported in the literature, we also hope that our study will contribute toward a more balanced understanding of the wider importance of this phenomenon.     

1H NMR studies of drugs with achiral and chiral lanthanide shift reagents: Applications to the anticonvulsant pheneturide

The anticonvulsant pheneturide, PNT, has been studied by 300 MHz ¹H NMR in CDCl₃ at ambient temperatures with the achiral lanthanide shift reagent (LSR) Eu(FOD)₃, and with the chiral LSR, Eu(HFC)₃. Both LSRs produced spectral simplification of the aryl proton signal region, and substantial lanthanide‐induced shifts (LIS). With added Eu(HFC)₃, enantiomeric shift differences (ΔΔδ) were induced for most nuclei of PNT, indicating substantial potential for direct determination of enantiomeric excess. Valley heights between corresponding signals in the PNT enantiomers as low as 3.6% were achieved for the meta resonance. Least squares line‐fitting was applied to the variation of chemical shift vs. [LSR]/[PNT] molar ratios for both LSRs. Tentative assignments were made for the NH absorptions based on two‐dimensional NMR (COSY45), as well as their relative magnitudes of LIS, ΔΔδ, and lanthanide‐induced line broadening. The PNT conformation reported in the crystal is believed to be retained in solution with added LSR. The relative senses of magnetic nonequivalence were found to be the same among the three sets of aryl protons, and among the three kinds of protons in the ethyl moiety, with high levels of added chiral LSR, using 2D NMR. Chirality 11:529–535, 1999. © 1999 Wiley‐Liss, Inc.     

No relationship between microsatellite variation and neonatal fitness in Antarctic fur seals, Arctocephalus gazella

Published studies of wild vertebrate populations have almost universally reported positive associations between genetic variation measured at microsatellite loci and fitness, creating the impression of ubiquity both in terms of the species and the traits involved. However, there is concern that this picture may be misleading because negative results frequently go unpublished. Here, we analyse the relationship between genotypic variation at nine highly variable microsatellite loci and neonatal fitness in 1070 Antarctic fur seal pups born at Bird Island, South Georgia. Despite our relatively large sample size, we find no significant association between three different measures of heterozygosity and two fitness traits, birth weight and survival. Furthermore, increasing genetic resolution by calculating parental relatedness also yields no association between genetic variation and fitness. Our findings are consistent with necropsy data showing that most pups die from starvation or trauma, conditions that are unlikely to be influenced strongly by genetic factors, particularly if the benefits of high heterozygosity are linked to immune-related genes.     

Heterozygosity-fitness correlations in the gilthead sea bream Sparus aurata using microsatellite loci from unknown and gene-rich genomic locations

Heterozygosity-fitness correlations (HFC) were assessed for a sample of a gilthead sea bream Sparus aurata population. Two hundred and seventy-one fish were genotyped at 22 known and novel microsatellite loci, from which correlations between the multilocus heterozygosity index (I(MLH) ) and various fitness traits (fork length, mass and specific growth rates) were calculated. Significant global HFCs were found in this sample (0·02 ≤r(2) ≤ 0·08). In addition, all the significant correlations found in this work were negative, indicating that heterozygotes had lower fitness than their homozygote counterparts. Marker location could not explain the observed HFCs. Evidence of inbreeding, outbreeding or population and family structuring was not found in this work. The presence of undetected general effects that may lead to the appearance of HFCs, however, cannot be ruled out. These results seem to be best explained by the occurrence of local effects (due to linkage) or even by possible direct locus advantages.     

LCA study of air conditioners with an alternative refrigerant

In the refrigeration and air conditioning industry, technologies to reduce environmental impacts, such as global warming, ozone-layer depletion, and discharging industrial wastes, are getting much attention nowadays. This paper reports the Life Cycle Assessment conducted to comparatively analyze two air conditioner units for residential use. One is a traditional type with HCFC22 being used for its refrigerant and the other is with HFC410A. Because the main focus of this analysis is on the comparison of the refrigerants, data of the refrigerants used are taken from the actual measurements in their production and disposal stages. As a result, the ozone layer depleting effect can be eliminated completely by using HFC410A. On the other hand, the global warming effect doesn’t get reduced extensively by using HFC410A. However, it does so by treating used refrigerants with a proper waste management. Moreover, it can be proved that using HFC410A reduces environmental impacts in all the other impact categories assessed, which are acidification, air pollution, water pollution, and energy consumption. To conclude this case study, replacing HCFC22 with HFC410A for the refrigerant is certainly one of the effective methods for reducing environmental impacts given by air conditioners.     

Heterozygosity–fitness correlations and associative overdominance: new detection method and proof of principle in the Iberian wild boar

Heterozygosity-fitness correlations (HFC) may result from a genome-wide process — inbreeding — or local effects within the genome. The majority of empirical studies reporting HFCs have attributed correlations to inbreeding depression. However, HFCs are unlikely to be caused by inbreeding depression because heterozygosity measured at a small number of neutral markers is unlikely to accurately capture a genome-wide pattern. Testing the strengths of localized effects caused by associative overdominance has proven challenging. In their current paper, Amos and Acevedo-Whitehouse present a novel test for local HFCs. Using stochastic simulations, they determine the conditions under which single-locus HFCs arise, before testing the strength of the correlation between the neutral marker and a linked gene under selection in their simulations. They used insights gained from simulation to statistically investigate the likely cause of correlations between heterozygosity and disease status using data on bovine tuberculosis infections in a wild boar population. They discover that a single microsatellite marker is an excellent predictor of tuberculosis progression in infected individuals. The results are relevant for wild boar management but, more generally, they demonstrate how single-locus HFCs could be used to identify coding loci under selection in free-living populations.     

A new test for genotype-fitness associations reveals a single microsatellite allele that strongly predicts the nature of tuberculosis infections in wild boar

There is increasing interest in the basis of commonly observed heterozygosity-fitness correlations (HFCs). Two models appear possible, a genome-wide effect due to inbreeding depression, and a single-locus effect due to chance linkage to a gene(s) experiencing balancing selection. Recent studies suggest that the latter tends to be more important in the majority of studies, but tests for the presence of single-locus effects tend to be rather weak. One of the problems is that the linkage disequilibrium between a microsatellite and a nearby gene experiencing balancing selection is never likely to be 100%. With this in mind, we conduct stochastic simulations aimed at determining the conditions under which single-locus HFCs may develop. We also suggest a new approach that could offer improved detection of HFCs but which also offers a more general method for detecting genotype-fitness correlations. Our method is based on looking for the maximum possible strength of association between genotype and fitness, and then asking whether randomized data sets are able to generate similarly strong associations. This method is tested on both simulated and real data. In both cases, our method generates greater levels of significance than current tests. Applied to previously published data from wild boar affected by tuberculosis, the method uncovers a strong single-allele association that is strongly predictive of whether the disease is localized or spreads throughout the body. We further suggest a simple method for dealing with the problem of population structure, and believe this approach will help to identify genomic regions associated with fitness.     

A panel of new microsatellite loci for genetic studies of antarctic fur seals and other otariids

Nine dinucelotide microsatellite loci were developed in the Antarctic fur seal Arctocephalus gazella. Each locus possessed between 4 and 9 alleles in a sample of twenty individuals sampled from Bird Island, South Georgia, and expected heterozygosity ranged from 0.52 to 0.84. All but one of the loci conformed to Hardy–Weinberg equilibrium and no evidence was found for genotypic disequilibrium. Additionally, all of the loci successfully cross-amplified in the South American fur seal Arctocephalus australis, the New Zealand fur seal Arctocephalus forsteri and the Steller’s sea lion Eumetopias jubatus, and six also yielded products in the more distantly related harbour seal Phoca vitulina and walrus Odobenus rosmarus. These loci should prove useful for studies of the population genetics of Antarctic fur seals and other important otariid species.     

SELECTION ON OVERDOMINANT GENES MAINTAINS HETEROZYGOSITY ALONG MULTIPLE CHROMOSOMES IN A CLONAL LINEAGE OF HONEY BEE

Correlations between fitness and genome‐wide heterozygosity (heterozygosity‐fitness correlations, HFCs) have been reported across a wide range of taxa. The genetic basis of these correlations is controversial: do they arise from genome‐wide inbreeding (“general effects”) or the “local effects” of overdominant loci acting in linkage disequilibrium with neutral loci? In an asexual thelytokous lineage of the Cape honey bee (Apis mellifera capensis), the effects of inbreeding have been homogenized across the population, making this an ideal system in which to detect overdominant loci, and to make inferences about the importance of overdominance on HFCs in general. Here we investigate the pattern of zygosity along two chromosomes in 42 workers from the clonal Cape honey bee population. On chromosome III (which contains the sex‐locus, a gene that is homozygous‐lethal) and chromosome IV we show that the pattern of zygosity is characterized by loss of heterozygosity in short regions followed by the telomeric restoration of heterozygosity. We infer that at least four selectively overdominant genes maintain heterozygosity on chromosome III and three on chromosome IV via local effects acting on neutral markers in linkage disequilibrium. We conclude that heterozygote advantage and local effects may be more common and evolutionarily significant than is generally appreciated.     

Secretome analysis using a hollow fiber culture system for cancer biomarker discovery

Secreted proteins, collectively referred to as the secretome, were suggested as valuable biomarkers in disease diagnosis and prognosis. However, some secreted proteins from cell cultures are difficult to detect because of their intrinsically low abundance; they are frequently masked by the released proteins from lysed cells and the substantial amounts of serum proteins used in culture medium. The hollow fiber culture (HFC) system is a commercially available system composed of small fibers sealed in a cartridge shell; cells grow on the outside of the fiber. Recently, because this system can help cells grow at a high density, it has been developed and applied in a novel analytical platform for cell secretome collection in cancer biomarker discovery. This article focuses on the advantages of the HFC system, including the effectiveness of the system for collection of secretomes, and reviews the process of cell secretome collection by the HFC system and proteomic approaches to discover cancer biomarkers. The HFC system not only provides a high-density three-dimensional (3D) cell culture system to mimic tumor growth conditions in vivo but can also accommodate numerous cells in a small volume, allowing secreted proteins to be accumulated and concentrated. In addition, cell lysis rates can be greatly reduced, decreasing the amount of contamination by abundant cytosolic proteins from lysed cells. Therefore, the HFC system is useful for preparing a wide range of proteins from cell secretomes and provides an effective method for collecting higher amounts of secreted proteins from cancer cells. This article is part of a Special Issue entitled: An Updated Secretome.