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Toxin-antitoxin ( TA) systems are small genetic elements found on plasmids or chromosomes of countless bacteria, archaea, and possibly also unicellular fungi. Under normal growth conditions, the activity of the toxin protein or its translation is counteracted by an antitoxin protein or noncoding RNA. Five types of TA systems have been proposed that differ markedly in their genetic architectures and modes of activity control. Subtle regulatory properties, frequently responsive to environmental cues, impact the behavior of TA systems. Typically, stress conditions result in the degradation or depletion of the antitoxin. Unleashed toxin proteins impede or alter cellular processes including translation, DNA replication, or ATP or cell wall synthesis. TA toxin activity can then result in cell death or in the formation of drug-tolerant persister cells. The versatile properties of TA systems have also been exploited in biotechnology and may aid in combating infectious diseases. [ABSTRACT FROM AUTHOR]

Quantitative trait loci (QTL) of longevity identified in human and mouse are significantly colocalized, suggesting that common mechanisms are involved. However, the limited number of strains that have been used in mouse longevity studies undermines the ability to identify longevity genes. We crossed C57BL/6J mice with a new wild-derived strain, Pohn, and identified two life span QTL—Ls1 and Ls2. Interestingly, homologous human longevity QTL colocalize with Ls1. We also defined new QTL for metabolic heat production and body weight. Both phenotypes are significantly correlated with life span. We found that large clone ratio, an in vitro indicator for cellular senescence, is not correlated with life span, suggesting that cell senescence and intrinsic aging are not always associated. Overall, by using Pohn mice, we identified new QTL for longevity-related traits, thus facilitating the exploration of the genetic regulation of aging. [ABSTRACT FROM PUBLISHER]

The strong association of the HLA-B27 with AS was first discovered independently by groups in London and California in 1972 and has recently been confirmed beyond reasonable doubt by fine mapping in the latest and most sophisticated genome-wide association study (GWAS) published this July. Yet, despite nearly four decades of extensive research, the exact role that HLA-B27 plays in pathogenesis remains unknown. However, we believe that recent developments in three fields have allowed us to view this conundrum in a new light and to propose coherent theories of disease pathogenesis. These areas are as follows: (i) GWASs, (ii) studies of B27 biology and (iii) lessons from biologic therapies. In this review we will discuss these recent advances before discussing the current models of AS pathogenesis under investigation. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate measures of boar (Sus scrofa) taint as potential selection criteria to reduce boar taint so that castration of piglets will become unnecessary. Therefore, genetic parameters of boar taint measures and their genetic correlations with finishing traits were estimated. In particular, the usefulness of a human panel assessing boar taint (human nose score) was compared with chemical assessment of boar taint compounds, androstenone, skatole, and indole. Heritability estimates for androstenone, skatole, and indole were 0.54, 0.41, and 0.33, respectively. The heritability for the human nose score using multiple panelists was 0.12, and ranged from 0.12 to 0.19 for individual panelists. Genetic correlations between scores of panelists were generally high up to unity. The genetic correlations between human nose scores and the boar taint compounds ranged from 0.64 to 0.999. The boar taint compounds and human nose scores had low or favorable genetic correlations with finishing traits. Selection index estimates indicated that the effectiveness of a breeding program based on human nose scores can be comparable to a breeding program based on the boar taint compounds themselves. Human nose scores can thus be used as a cheap and fast alternative for the costly determination of boar taint compounds, needed in breeding pigs without boar taint. [ABSTRACT FROM AUTHOR]

Sorbitol-fermenting Escherichia coli O157: NM ( SF O157) is an emerging pathogen suggested to be more virulent than nonsorbitol-fermenting Escherichia coli O157: H7 ( NSF O157). Important virulence factors are the Shiga toxins ( stx), encoded by stx1 and/or stx2 located within prophages integrated in the bacterial genome. The stx genes are expressed from p R′ as a late protein, and anti-terminator activity from the Q protein is necessary for read through of the late terminator t R′ and activation of p R′. We investigated the regulation of stx2EDL933 expression at the genomic level in 17 Norwegian SF O157. Sequencing of three selected SF O157 strains revealed that the anti-terminator q gene and genes upstream of stx2EDL933 were identical or similar to the ones observed in the E. coli O111: H− strain AP010960, but different from the ones observed in the NSF O157 strain EDL933 ( AE005174). This suggested divergent stx2EDL933 -encoding bacteriophages between NSF O157 and the SF O157 strains (). Furthermore, different DNA structures were detected in the SF O157 strains, suggesting diversity among bacteriophages also within the SF O157 group. Further investigations are needed to elucidate whether the qO111:H − gene observed in all our SF O157 contributes to the increased virulence seen in SF O157 compared to NSF O157. An assay for detecting qO111:H − was developed. [ABSTRACT FROM AUTHOR]

The inverted teat defect is characterized by the failure of teats to protrude from the udder surface and has a negative effect on the economic efficiency of pig production. The inverted teat defect is influenced by genetic factors, but the number and identity of relevant genes are unknown. In this study, we compared the mRNA expression of teat tissues from unaffected pigs and affected pigs by using microarrays. Simultaneously, 24,123 probe sets were screened, of which some 15,000 had present calls and were analyzed for differential expression between mesenchymal and epithelial tissue of 3 categories of teats (i.e., normal teats of unaffected and affected animals, and inverted teats of the latter). Differential expression was more pronounced in epithelial than in mesenchymal tissue, and the comparisons among the 3 categories of teats showed that local processes at the side of the affected area as well as processes taking place at the level of the organ contribute to the development of inverted teats. Genes related to biofunctions of cell maintenance, proliferation, differentiation, and replacement; organismal, organ, and tissue development; genetic information and nucleic acid processing; and cell-to-cell signaling and interaction were differentially expressed, depending on the teat phenotype and the status of the animal as affected or unaffected. In particular, genes encoding members of canonical pathways of growth factor signaling were highlighted. Complementary to previous real-time quantitative reverse-transcription PCR experiments showing upregulation of growth factors (epidermal growth factor, fibroblast growth factor, hepatocyte growth factor, platelet-derived growth factor, vascular endothelial growth factor) and their receptors in the inverted teat, here it is shown that the abundance of transcripts encoding subordinated proteins (acid phosphatase 1, soluble; activating transcription factor 2; casein kinase 2, α 1 polypeptide; casein kinase 2, α prime polypeptide; actinin, α 2; and Homo sapiens growth factor receptor-bound protein 2) within the growth factor signaling pathways are also affected. Tuning of the expression of genes of these pathways balances the differentiation and proliferation of epithelial and mesenchymal teat tissue and finally affects the shape and structure of the teats [ABSTRACT FROM AUTHOR]

Objective. A large number of genes, including several not previously implicated in SLE susceptibility, have recently been identified or confirmed by genome-wide association studies (GWAS). In this study, we sought to replicate some of these results in Finnish SLE patients (n = 275) and control individuals (n = 356).Methods. We genotyped 32 single nucleotide polymorphisms (SNPs) in 12 of the best-supported GWAS-identified SLE genes and loci. We further investigated gene–gene interactions between the loci included in the study.Results. The strongest evidence of association was found at the IRF5-TNPO3 locus, with the most significant P-value being 2.0 × 10−7 and an odds ratio of 1.95 (95% CI 1.51, 2.50). Association between SLE and TNFAIP3, FAM167A-BLK, BANK1 and KIAA1542 was also confirmed, although at a lower significance level and contribution to individual risk. No significant association was found with 1q25.1, PXK, ATG5, ICA1, XKR6, LYN and SCUBE1. Furthermore, no significant gene–gene interactions were detected.Conclusion. Replication of previous GWAS findings across diverse populations is of importance to validate these associations and to get a better understanding of potential genetic heterogeneity between populations in SLE susceptibility. Our results attest the importance of B-cell receptor pathway and IFN signalling in SLE pathogenesis. [ABSTRACT FROM PUBLISHER]

We have investigated whether the quality of dietary fat and supplementation with coenzyme Q10 (CoQ) modifies expression of genes related with inflammatory response and endoplasmic reticulum stress in elderly persons. Twenty participants received three diets for 4 weeks each: Mediterranean diet + CoQ (Med + CoQ), Mediterranean diet (Med), and saturated fatty acid–rich diet (SFA). After 12-hour fast, volunteers consumed a breakfast with a fat composition similar to that consumed in each of the diets. Med and Med + CoQ diets produced a lower fasting calreticulin, IL-1b, and JNK-1 gene expression; a lower postprandial p65, IKK-b, MMP-9, IL-1b, JNK-1, sXBP-1, and BiP/Grp78 gene expression; and a higher postprandial IkB-a gene expression compared with the SFA diet. Med + CoQ diet produced a lower postprandial decrease p65 and IKK-b gene expression compared with the other diets. Our results support the anti-inflammatory effect of Med diet and that exogenous CoQ supplementation in synergy with a Med diet modulates the inflammatory response and endoplasmic reticulum stress. [ABSTRACT FROM PUBLISHER]

Objectives. To determine whether increased levels of B-cell activating factor (BAFF) in patients with SLE are due to disease activity or genetic variations in the promoter region of the BAFF gene and BAFF gene expression.Methods. The case–control study included 101 SLE patients and 111 healthy controls. Five single nucleotide polymorphisms (SNPs) in the BAFF promoter region were investigated by melting point analysis: c.-2841 (T > C), c.-2704 (T > C), c.-2701 (A > T), c.-871 (C > T) and c.-514 (A > G). BAFF mRNA levels were determined by real-time PCR (BAFF-RQ) and serum BAFF (s-BAFF) levels were measured by ELISA. Independent predictors that might be correlated with increased s-BAFF in SLE patients were analysed by multivariate regression methods.Results. Although s-BAFF levels were increased in SLE patients (1.73 vs 0.98 ng/μl, P < 0.001), no specific BAFF genotype was found to associate with SLE. The different genotypes defined by the investigated SNPs were identified both in SLE patients and healthy controls with similar frequencies. No association was found between BAFF genotype and BAFF-RQ. s-BAFF was independent of other factors, correlated with CRP (β = 0.40, P < 0.001) and physician's visual analogue score (R = 0.21, P = 0.046) and inversely with haemoglobin (β = −0.32, P < 0.001) and IgA (β = −0.33, P = 0.001).Conclusions. Increased s-BAFF levels in SLE patients are associated with the acute-phase responses, CRP and haemoglobin, but probably not dependent on BAFF genotype or expression. This indicates that s-BAFF production occurs at sites of inflammation. [ABSTRACT FROM PUBLISHER]

Within the last year, all 50 states in the United States have adopted newborn screening (NBS) protocols for cystic fibrosis (CF), the most common fatal autosomal recessive disease among Caucasian populations. In this overview, we discuss the rationale for implementing NBS for CF and discuss the different testing algorithms states have adopted. Based on studies in the United States, Australia, and the United Kingdom, these measures will likely lead to less severe disease, prolonged life, and more cost-effective management of CF in the long run. [ABSTRACT FROM AUTHOR]

Objectives. CD38 participates in lymphocyte ontogeny and function and may be involved in autoimmunity. Absence of CD38 accelerates development of non-obese diabetic (NOD) mice diabetes and anti-CD38 antibodies are good markers of human disease. Little is known regarding systemic autoimmunity. Active SLE patients have higher numbers of CD38+ T and B cells. CD38 is a candidate gene for the murine Lmb2 lupus locus. We aimed to investigate whether CD38 was involved in lupus development.Methods. We developed Cd38−/−-Faslpr/Faslpr mice and monitored them for development of a lupus-like disease through measurement of protein excretion in urine, histological assessment of the kidneys, quantification of circulating immunoglobulins and autoantibodies. We have also immunophenotyped 2- and 6-month old Cd38−/−-Faslpr/Faslpr mice.Results. We found that absence of CD38 accelerated disease development: female Cd38−/−-Faslpr/Faslpr mice presented severe proteinuria, GN, deposition of ICs in the renal medulla and increased amounts of circulating immunoglobulin G (IgG), although anti-dsDNA autoantibodies and RF were not significantly increased at disease onset. We have found that Cd38−/−-Faslpr/Faslpr male mice, similarly to other murine models of lupus, were able to control disease. Absence of CD38 in lpr mice altered differentiation of T cells and dendritic cells (DC).Conclusion. Although the role of CD38 in tolerance is still to be elucidated, we provide evidence that it may play an active role in the control of a murine lupus-like disease. [ABSTRACT FROM PUBLISHER]

Despite vaccinations, hepatitis B virus (HBV) infections are still very common worldwide. The virus replicates by reverse transcription using a viral polymerase lacking proof-reading ability. This results in the emergence of mutant viruses that can be selected out by host immunity or viral therapeutic agents. Several well-characterized HBV variants have been identified that challenge the effectiveness of the current vaccines. Other mutations result in a change within the HBV surface antigen, resulting in a loss of detection by some diagnostic assays. Additionally, a number of mutations have evolved in response to antiviral therapy. This report is an overview of the HBV mutations leading to vaccine failure, loss of HBV detection by diagnostic assays, increased viral replication, and resistance to antiviral agents. [ABSTRACT FROM AUTHOR]

Objectives. To investigate whether maintaining the chondrocyte’s native pericellular matrix prevents collagen-induced up-regulation of collagenase-3 (MMP-13) and whether integrin α1 (ITGα1) and/or discoidin domain receptor 2 (DDR2) modulate MMP-13 expression and which signalling pathway plays a role in collagen-stimulated MMP-13 expression.Methods. Goat articular chondrocytes and chondrons were cultured on collagen coatings. Small interfering RNA (siRNA) oligonucleotides targeted against ITGα1 and DDR2 were transfected into primary chondrocytes. Chemical inhibitors for mitogen-activated protein kinase kinase (MEK1) (PD98059), focal adhesion kinase (FAK) (FAK inhibitor 14), mitogen-activated protein kinase 8 (JNK) (SP600125) and protein kinase C (PKC) (PKC412), and a calcium chelator (BAPTA-AM) were used in cell cultures. Real-time PCR was performed to examine gene expression levels of MMP-13, ITGα1 and DDR2 and collagenolytic activity was determined by measuring the amount of hydroxyproline released in the culture medium.Results. Maintaining the chondrocyte’s native pericellular matrix prevented MMP-13 up-regulation and collagenolytic activity when the cells were cultured on a collagen coating. Silencing of ITGα1 and DDR2 reduced MMP-13 gene expression and collagenolytic activity by primary chondrocytes cultured on collagen. Incubation with the PKC inhibitor strongly reduced MMP-13 gene expression levels. Gene expression levels of MMP-13 were also decreased by chondrocytes incubated with the MEK, FAK or JNK inhibitor.Conclusion. Maintaining the native pericellular matrix of chondrocytes prevents collagen-induced up-regulation of MMP-13. Both ITGα1 and DDR2 modulate MMP-13 expression after direct contact between chondrocytes and collagen. PKC, FAK, MEK and JNK are involved in collagen-stimulated expression of MMP-13. [ABSTRACT FROM PUBLISHER]

The integrity of the vascular system is essential for the efficient functioning of the brain. Aging-related structural and functional disturbances in the macro- or microcirculation of the brain make it vulnerable to cognitive dysfunction, leading to brain degeneration and dementing illness. Several faltering controls, including impairment in autoregulation, neurovascular coupling, blood-brain barrier leakage, decreased cerebrospinal fluid, and reduced vascular tone, appear to be responsible for varying degrees of neurodegeneration in old age. There is ample evidence to indicate vascular risk factors are also linked to neurodegenerative processes preceding cognitive decline and dementia. The strongest risk factor for brain degeneration, whether it results from vascular or neurodegenerative mechanisms or both, is age. However, several modifiable risks such as cardiovascular disease, hypertension, dyslipidemia, diabetes, and obesity enhance the rate of cognitive decline and increase the risk of Alzheimer's disease in particular. The ultimate accumulation of brain pathological lesions may be modified by genetic influences, such as the apolipoprotein E ε4 allele and the environment. Lifestyle measures that maintain or improve cardiovascular health, including consumption of healthy diets, moderate use of alcohol, and implementation of regular physical exercise are important factors for brain protection. [ABSTRACT FROM AUTHOR]

Information regarding several papers discussed at a symposium on the functions and adaptations of the human brain and its relation with age is presented. Topics include the receptor biology of the brain, the impact of environmental factors into the brain, and the secular changes of the brain in aging. The symposium featured several international nutrition experts including Tom Kirkwood, Marshal Folstein, and Richard Wurtman.

Intrinsic aging of the brain and its relationship to age-related neurodegenerative diseases need to be understood as part of global aging, which results from gradual accumulation of a variety of kinds of cellular and molecular damage. While certain kinds of molecular lesions are particularly associated with specific diseases, there is substantial overlap between the pathways causing disease and those contributing to global aging. Growing evidence indicates that the global aging process is more malleable than used to be thought. This needs to be taken into account in efforts to improve health and retention of mental capital across the life course. [ABSTRACT FROM AUTHOR]

This article reviews developmental processes in the human brain and basic principles underlying typical and atypical motor development. The Neuronal Group Selection Theory is used as theoretical frame of reference. Evidence is accumulating that abundance in cerebral connectivity is the neural basis of human behavioral variability (ie, the ability to select, from a large repertoire of behavioral solutions, the one most appropriate for a specific situation). Indeed, typical human motor development is characterized by variation and the development of adaptive variability. Atypical motor development is characterized by a limited variation (a limited repertoire of motor strategies) and a limited ability to vary motor behavior according to the specifics of the situation (ie, limited variability). Limitations in variation are related to structural anomalies in which disturbances of cortical connectivity may play a prominent role, whereas limitations in variability are present in virtually all children with atypical motor development. The possible applications of variation and variability in diagnostics in children with or at risk for a developmental motor disorder are discussed. [ABSTRACT FROM AUTHOR]

Several combinations of genes encoding KIR3DL1 alleles and their HLA-Bw4 ligands have been linked with favorable outcomes upon exposure to or infection with human immunodeficiency virus (HIV). Some protective KIR3DL1/HLA-Bw4 combinations confer elevated natural killer (NK) cell functional potential. The K562-stimulated functionality of NK cells from KIR3DL1*004/HLA-Bw4 and control genotype carriers was assessed by flow cytometry and found to be higher in KIR3DL1*004/HLA-Bw4 carriers. However, a comparison of the frequency of this combined genotype among HIV-exposed uninfected and HIV-infected subjects revealed no between-group differences. Thus, despite its ability to license NK cells, KIR3DL1*004/HLA-Bw4 is not associated with a reduced risk of infection. [ABSTRACT FROM AUTHOR]

The tendency to fight is a well-known behavior in Valdostana cattle, and noncruel traditional contests are organized yearly by farmers to identify the most dominant cow. Cow battles consist of elimination matches that have important economic implications for both tourism and farmers. The aims of this study are 1) to validate a scoring system to express fighting ability, and 2) to carry out a genetic analysis for this trait using different data sets and models. A data set accounting for 16,509 fighting records of 5,981 cows relevant to contests over 6 yr was retained after editing (data set 1). Data on placements were used to compute a placement score accounting for wins, tournament size, and difficulty, and differentiating the 20 preliminary battles each year from the final match. A second data set was created using only the individual best yearly placement scores, that is, deleting repeats with a smaller placement score for the same animal within each year (data set 2; n 10,367 records, corresponding to a single datum per year per cow). Compared with the placement or position of each cow, the placement score proved to be less skewed (-1.45 for placement position and 1.25 for placement score, respectively) and exhibited better coefficients for the probability of a normal distribution. An animal model REML method analysis (accounting for 13,456 animals in the pedigree) was carried out, with consideration given to different combinations of fixed and random nongenetic factors other than the random animal and permanent environmental effects. Results indicated that random factors other than additive genetic and permanent environment effects did not improve the model fit; therefore, it was not useful to take them into account. Heritability estimates obtained with the model showing the best fit were 0.078 (data set 1) and 0.098 (data set 2). Results of this study indicate that selection for fighting ability in Valdostana cattle using data on battle performance is possible. [ABSTRACT FROM AUTHOR]

Apoptosis is a form of cell death that involves the changes of mitochondrial function and the regulated activation of caspase cascades, which selectively cleave cytoskeleton proteins and catalyze the changes of cell organelles and morphological structure. The changes of mitochondrial function, cell morphological structure, and degradation of cytoskeleton are considered to be responsible for the development of meat qualities. The LM, semitendinosus, and psoas minor (PM,) muscles of 5 crossbred bulls were used to observe the morphologic and quantitative changes of apoptosis, as well as the change of caspase-3 activity during 7 d storage. Transmission electron microscopy shows that the typical features of apoptosis appeared in muscles between d 1 and 4. Terminal deoxynucleotidyl transferase-mediated deoxyuricline triphosphate nick end-labeling (TUNEL) positive nuclei were detected at d 4 and increased subsequently. The count of TUNEL-positive nuclei was different in 3 muscles at d 7 (P < 0.001). There was a significant increase in caspase-3 activities at 4 h postmortem relative to the activities at the first 30 mm in 3 muscles (P 0.0147 in LM; P = 0.0058 in PM; P = 0.0306 in sernitendinosus), and the apexes had 2.9 to 6.5 times more activities than activities at the first 30 mm postmortem. Apoptosis did exist in 3 types of muscles during the conditioning period. Apoptosis and caspase cascades system could be associated with the postmortem development of meat quality in skeletal muscles of bulls. [ABSTRACT FROM AUTHOR]

BACKGROUND: The relationship between fibroids and infertility remains an unsolved question, and management of intramural fibroids is controversial. During the implantation phase, uterine peristalsis is dramatically reduced, which is thought to facilitate embryo implantation. Our aims were to evaluate (i) the occurrence and frequency of uterine peristalsis in infertile women with intramural fibroids and (ii) whether the presence of uterine peristalsis decreases the pregnancy rate. [ABSTRACT FROM AUTHOR]

The aim of this study was to estimate genetic parameters for pork intramuscular fatty acid (FA) composition and indices for desaturase and elongase activities involved in n-3 and n-6 PUFA metabolism. The LM of 437 slaughter pigs was analyzed for FA composition (expressed as g/100 g of FA). Indices for enzyme activities were calculated from product to precursor FA ratios. Genetic parameters were estimated with singleand multi-trait animal models. The total FA content, reflecting the intramuscular fat content, was either included or not in the model. Results from the models without total FA content showed relatively high heritability estimates, generally above 0.50, for the proportion of the most important MUFA and PUFA, compared with much smaller values for the SFA. When total FA content was included in the models, heritability values decreased (P < 0.001) for most individual FA and for all sums of FA groups, except for C18:0, C18:3n-6, and C18:3n-3. Heritability estimates for the ratios C20:4n-6/C18:2n-6 and C22:6n-3/C18:3n-3, reflecting the overall conversion in the n-6 and n-3 PUFA pathway, respectively, were 0.29 and 0.35, respectively, with total FA content in the model and increased to 0.38 and 0.49, respectively, if total FA content was not in the model. Heritabilities for other more specific indices were of the same order. Genetic correlations between PUFA proportions and indices for enzyme activities with ADG were mostly negative, whereas the correlations with carcass lean meat percentage were mostly positive. It was concluded that there is meaningful genetic variation for long-chain PUFA metabolism that is only partly dependent on the carcass and muscle fat content. This may allow selection for improved FA composition of pork. [ABSTRACT FROM AUTHOR]

Little is known about mutational variability in livestock, among which only a few mutations with relatively large effects have been reported. In this manuscript, mutational variability was analyzed in 1,765 litter size records from 404 Ripollesa ewes to characterize the magnitude of this genetic source of variation and check the suitability of including mutational effects in genetic evaluations of this breed. Threshold animal models accounting for additive genetic mutations were preferred to models without mutational contributions, with an average difference in the deviance information criterion of more than 5 units. Moreover, the statistical relevance of the additive genetic mutation term was checked through a Bayes factor approach, which showed that the models with mutational variability were 8.5 to 22.7 times more probable than the others. The mutational heritability (percentage of the phenotypic variance accounted for by mutational variance) was 0.6 or 0.9%, depending on whether genetic dominance effects were accounted for by the analytical model. The inclusion of mutational effects in the genetic model for evaluating litter size in Ripollesa ewes called for some minor modifications in the genetic merit order of the individuals evaluated, which suggested that the continuous uploading of new additive mutations could be taken into account to optimize the selection scheme. This study is the first attempt to estimate mutational variances in a livestock species and thereby contribute to better characterization of the genetic background of productive traits of interest. [ABSTRACT FROM AUTHOR]

Debate surrounding the integration of phenotypic plasticity within the neo-Darwinian paradigm has recently intensified, but is largely dominated by conceptual abstractions. Advances in our capacities to identify candidate genes, and quantify their levels of expression, now facilitate the study of natural variation in inherently plastic traits, and may lead to a more concrete understanding of plasticity's role in adaptive evolution. We present data from parapatric threespine stickleback ( Gasterosteus aculeatus) demes inhabiting geologically recent, freshwater and saltwater zones of a large estuary. Reaction norms for survival confirm adaptation to local salinity conditions. Analysis of osmoregulatory candidate gene expression within an ecological quantitative genetics framework suggests putative mechanisms underlying adaptive variation, and provides insights into the role of ancestral trait plasticity in this divergence. A sodium–potassium ATPase (ATP1A1) is identified as a candidate gene for freshwater adaptation. In addition to heritable variation for gene expression, we infer significant correlation between measures of expression and individual fitness. Overall results indicate a loss of plasticity in the freshwater deme. We discuss how this is consistent with adaptation facilitated by ancestral plasticity as a heuristic example that may prove useful for future, explicit tests of the genetic assimilation hypothesis. [ABSTRACT FROM AUTHOR]

Background Epidermolytic ichthyosis (EI), previously termed bullous congenital ichthyosiform erythroderma or epidermolytic hyperkeratosis, is a clinically heterogeneous genodermatosis caused by mutations in the genes encoding the suprabasal keratins 1 and 10. Classical EI is clinically characterized by severe neonatal erythroderma, blistering and fragile skin in infancy, quickly subsiding with subsequent development of generalized scaling hyperkeratosis. We report three Dutch families with palmoplantar keratoderma and mild blistering, but without neonatal erythroderma and generalized scaling. A novel heterozygous missense mutation in the linker L12 domain of KRT1:c.1019A>G, p.Asp340Gly was found associated with this phenotype in these families. Objectives To investigate the effects of the novel KRT1:p.Asp340Gly and the one other previously reported KRT1:p.Asp340Val mutations on keratinocyte cytoskeleton formation and stress resistance. Methods Wild-type and mutant pEGFP- KRT1 fusion constructs were transfected into HaCaT cells and exposed to hypo-osmotic shock. Haplotyping and genealogical studies were performed to investigate the possibility of a common founder for p.Asp340Gly. Results Cells transfected with either one of the keratin 1 L12 domain mutations showed significantly increased tonofilament aggregation. The haplotype around the KRT1 gene was shared in all affected family members of two families and a common founder was traced. Conclusions Our study supports the pathogenicity of the keratin 1 L12 domain mutations in vitro. These mutations are associated with a milder EI phenotype with pronounced palmoplantar keratoderma, and without neonatal erythroderma and scaling. The KRT1:p.Asp340Gly mutation in the Dutch families is likely to have arisen from a common founder. [ABSTRACT FROM AUTHOR]

The lysosomal protease cathepsin D is likely involved in β-amyloidogenesis in Alzheimer’s disease (AD). There is evidence for a single nucleotide polymorphism (rs17571) of the cathepsin D gene to be associated with increased AD risk. However, little is known about gender-specific differences. Therefore, we performed a genetic association study focusing on gender-specific differences in 434 participants (219 AD and 215 controls). Screening of the rs17571 shows a significantly higher proportion of T-allele carriers among male Alzheimer patients (28.5%) when compared with male controls (13.8%, p = .013, pcorr = .039). The odds ratio was 2.48 (95% confidence interval: 1.14–5.58). There was no significant difference in the T-allele distribution in women. Including APOE4 status and age did not have an additional effect on the morbidity risk. Thus, our results support the idea that rs17571 confers an increased risk for AD in men but not in women. Further investigation should substantiate the role of gender for AD risk of rs17571. [ABSTRACT FROM PUBLISHER]

Cell division depends on the correct localization of the cyclin-dependent kinases that are regulated by phosphorylation, cyclin proteolysis, and protein-protein interactions. Although immunological assays can define cell cycle protein abundance and localization, they are not suitable for detecting the dynamic rearrangements of molecular components during cell division. Here, we applied an in vivo approach to trace the subcellular localization of 60 Arabidopsis (Arabidopsis thaliana) core cell cycle proteins fused to green fluorescent proteins during cell division in tobacco (Nicotiana tabacurn) and Arabidopsis. Several cell cycle proteins showed a dynamic association with mitotic structures, such as condensed chromosomes and the preprophase band in both species, suggesting a strong conservation of targeting mechanisms. Furthermore, colocalized proteins were shown to bind in vivo, strengthening their localization-function connection. Thus, we identified unknown spatiotemporal territories where functional cell cycle protein interactions are most likely to occur. [ABSTRACT FROM AUTHOR]

Sequencing of the human genome has established that our DNA harbours many endogenous retrovirus (ERV) sequences, remnants of ancestral exogenous retroviral infections fixed in the germline DNA. In recent years, human ERVs (HERVs) have been implicated in melanomagenesis. Retrovirus-like particles and the expression of HERV mRNA and proteins have been demonstrated in melanoma tissue. In addition, antibodies to HERV proteins have been observed in patients with melanoma. In vitro and mouse models have provided fascinating insights into the potential mechanisms of HERVs in melanomagenesis. This review considers the evidence associating HERVs with melanoma. [ABSTRACT FROM AUTHOR]

Two genetic lines of barrows and gilt pigs with lean BW gain averages of 280 and 375 g/d were used to evaluate their macro- and micromineral contents at BW intervals from 20 to 125 kg of BW. The experiment was a 2 × 2 × 5 factorial arrangement of treatments (i.e., 2 sexes, 2 genetic lines, and 5 BW intervals) conducted in a completely randomized design in 6 replicates using a total of 120 pigs. Initially, 12 pigs (3 from each generic line and sex) were killed, and then at approximately 25 kg of BW intervals to 125 kg. Pigs were fed vitamin and mineral fortified corn-soybean meal diets. At slaughter the total body (except digesta and blood) of each pig was ground and analyzed for their macro- and micromineral contents. The high-lean genetic line (P <0.03) pigs and barrows (P < 0.01) reached their targeted BW an average 3 d earlier than the low-lean genetic line and guts. Total macro- and micromineral contents increased as BW increased, generally in a linear on' quadratic (P < 0.01) manner. There was an increasing difference between genetic lines in some minerals as BW increased. Total body Ca content was greater in the low-lean genetic line with increasing differences occurring as BW increased resulting in a BW × genetic line interaction (P < 0.05), whereas P was similar for both genetic lines. The quantity of K (P < 0.01) and S (P < 0.01) increased at a greater rate in the high-lean genetic line as BW increased, resulting in BW × genetic line interactions (P < 0.01). Body Cl (P < 0.01), Mg (P < 0.06), Mn (P < 0.05), Se (P < 0.01), and Zn (P < 0.01) were greater in the high-lean genetic. As BW increased, the Ca:P and the P:K ratios were increasingly greater (P < 0.01) in the low-lean genetic line, whereas the K:Na ratio was greater (P < 0.01) in high-lean genetic line. Although K and Fe were greater (P < 0.05) in gilts than in barrows, other mineral content differences were not significant. When minerals were expressed on a per kilogram of empty BW basis, the macro- and microminerals differed (P < 0.01) as BW increased indicating a response by body maturity. Genetic line had a greater effect on mineral content per kilogram of empty BW than sex. These results indicate that differences in mineral content are largely affected by BW or physiological age and by genetic line. Best-fitting equations were developed to determine macro- and micromineral contents of both genetic lines. [ABSTRACT FROM AUTHOR]

OBJECTIVE: 17-Hydroxylase/17,20-lyase deficiency (17OHD) results from mutations in the CYP17A1 gene, leading to failure to synthesize cortisol, adrenal androgens, and gonadal steroids. Adrenarche is a consequence of the increased production of adrenal androgens. Here, we report a case carrying novel R239Q mutation causing complete functional loss of CYP17A1, and thus absence of adrenal and gonadal sex hormone production. The patient has had unexpected pubic hair development and insufficient breast development with estrogen replacement therapy. Possible mechanisms leading to pubic hair development and breast underdevelopment are discussed. PATIENT AND METHODS: A 15-year-old female born to consanguineous parents presented with the lack of full breast development and irregular menses after the age of 14 years. She had Tanner III breast development on one side, Tanner I on the other side and Tanner I pubic hair and, no axillary hair development. The serum levels of FSH, LH, and progesterone were high and, estradiol was low. The measurement of basal and ACTH-stimulated steroids was consistent with the diagnosis of 17OHD. Genetic analysis revealed novel homozygous mutation R239Q in CYP17A1 gene. Therapy with hydrocortisone was initiated and followed by the addition of conjugated estrogen. Her breast development did not improve considerably, however, pubic hair development started after estrogen treatment in spite of undetectable serum levels of androgens. CONCLUSION: This case study suggests that estrogen exerts a permissive effect on pubic hair development in girls, even in the presence of very low-circulating androgens, and impaired breast development might be due to estrogen/progesterone imbalance in breast tissue. [ABSTRACT FROM AUTHOR]

It is generally assumed that familial aggregation of lipids relates to both genetic and shared environmental factors. To determine the degree to which familial similarities in lifestyle habits explain familial aggregation of high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol, the authors analyzed 1994–1996 data from 2,284 US adult participants in the National Heart, Lung, and Blood Institute Family Heart Study. For men and women, respectively, HDL cholesterol correlated with alcohol consumption (r= 0.27, p < 0.001; r = 0.21, p < 0.001), exercise (r = 0.06, p = 0.05; r = 0.10, p = 0.002), and smoking (r = −0.09, p = 0.005; r = −0.13, p < 0.001). There was strong familial aggregation of HDL cholesterol (parent-child, r = 0.32; sibling-sibling, r = 0.29), but less than 10% was explained by lifestyle habits. For LDL cholesterol, weak correlations were found for intake of total fat (r = 0.06, p = 0.07) and fruits/vegetables (r = −0.09, p = 0.005) among men and for smoking (r = 0.10, p = 0.002) among women. LDL cholesterol correlated strongly among family members (parent-child, r = 0.24; sibling-sibling, r = 0.31), but essentially none of this aggregation related to the lifestyle factors studied. This study suggests that lifestyle factors have little effect on the familial aggregation of HDL and LDL cholesterol. Am J Epidemiol 1999;150:910-18. [ABSTRACT FROM PUBLISHER]

Objective. The purpose of this study was to examine the association of the alpha-actinin-3 (ACTN3) R577X polymorphism on muscle function and physical performance in older adults. Methods. We measured knee extensor torque, midthigh muscle cross-sectional area, muscle quality, short physical performance battery score, and 400-meter walk time at baseline and after 5 years in white older adults aged 70-79 years in the Health, Aging and Body Composition Study cohort (n = 1367). Incident persistent lower extremity limitation (PLL) over 5 years was additionally assessed. We also examined white men in the Osteoporotic Fractures in Men Study, a longitudinal, observational cohort (n = 1152) of men 65 years old or older as a validation cohort for certain phenotypes. Results. There were no significant differences between genotype groups in men or women for adjusted baseline phenotypes. Male X-homozygotes had a significantly greater adjusted 5-year increase in their 400-meter walk time compared to R-homozygotes and heterozygotes (p = .03). In women, X-homozygotes had a ∼-35% greater risk of incident PLL compared to R-homozygotes (hazard ratio = 0.65, 95% confidence interval = 0.44-0.94). There were no other significant associations between any of the phenotypes and ACTN3 genotype with aging in either cohort. Conclusions. The ACTN3 polymorphism may influence declines in certain measures of physical performance with aging in older white adults, based on longitudinal assessments. However, the influence of the ACTN3 R577X polymorphism does not appear to have a strong effect on skeletal muscle-related phenotypes based on the strength and consistency of the associations and lack of replication with regard to specific phenotypes. [ABSTRACT FROM AUTHOR]

Clinally varying traits in Drosophila melanogaster provide good opportunities for elucidating the genetic basis of adaptation. Resistance to ethanol, a natural component of D. melanogaster's breeding sites, increases with latitude on multiple continents, indicating that the trait is under selection. Although the well-studied Alcohol dehydrogenase ( Adh) polymorphism makes a contribution to the clines, it accounts for only a small proportion of the phenotypic variation. We describe an amino acid replacement polymorphism in Aldehyde dehydrogenase ( Aldh), the gene encoding the second enzyme in the ethanol degradation pathway, that shows hallmarks of also contributing to the clines. The derived Aldh allele, like the Adh-Fast allele, increases in frequency in laboratory populations selected for ethanol resistance, and increases in frequency with latitude in wild populations. Moreover, strains with the derived allele have significantly higher ALDH enzyme activity with acetaldehyde (the breakdown product of ethanol) as a substrate than strains with the ancestral allele. As is the case with the Adh-Fast allele, chromosomes with the derived Aldh allele show markedly reduced molecular variation in the vicinity of the replacement polymorphism compared to those with the ancestral allele, suggesting a single, relatively recent origin. Nonetheless, the Aldh polymorphism differs from the Adh polymorphism in that the ethanol-associated allele remains in relatively low frequency in most populations. We present evidence that this is likely to be the result of a trade-off in catalytic activity, with the advantage of the derived allele in acetaldehyde detoxification being offset by a disadvantage in detoxification of other aldehydes. [ABSTRACT FROM AUTHOR]

Medicago truncatula has become a model system to study legume biology. It is imperative that detailed growth characteristics of the most commonly used cultivar, line A17 cv Jemalong, be documented. Such analysis creates a basis to analyze phenotypic alterations due to genetic lesions or environmental stress and is essential to characterize gene function and its relationship to morphological development. We have documented morphological development of M. truncatula to characterize its temporal developmental growth pattern; developed a numerical nomenclature coding system that identifies stages in morphological development; tested the coding system to identify phenotypic differences under phosphorus (P) and nitrogen (N) deprivation;. and created visual models using the L-system formalism. The numerical nomenclature coding system, based on a series of defined growth units, represents incremental steps in morphological development. Included is a decimal component dividing growth units into nine substages. A measurement component helps distinguish alterations that may be missed by the coding system. Growth under N and P deprivation produced morphological alterations that were distinguishable using the coding system and its measurement component. N and P deprivation resulted in delayed leaf development and expansion, delayed axillary shoot emergence and elongation, decreased leaf and shoot size, and altered root growth. Timing and frequency of flower emergence in P-deprived plants was affected. This numerical coding system may be used as a standardized method to analyze phenotypic variation in M. truncatula due to nutrient stress, genetic lesions, or other factors and should allow valid growth comparisons across geographically distant laboratories. [ABSTRACT FROM AUTHOR]

The article offers information on Equus asinus or wild ass, including its diagnosis, general characteristics, distribution, fossil record, form and function, ontogeny and reproduction, behavior, genetics, and conservation status. The authors believe that the first domestication of wild ass, based on morphology, may have occurred either in Near East or based on historical evidence, in northeastern Egypt or northwestern Africa. However, its oldest specimen is claimed to be from Syria which was dated to 9,000 B.C. In addition, the large populations of feral Equus asinus exist in the western U.S.

Two malic enzyme alleles, Men113A and Men113G, occur at approximately equal frequency in North American populations of Drosophila melanogaster, while only Men113A occurs in African populations. We investigated the population genetics, biochemical characteristics, and selective potential of these alleles. Comparable levels of nucleotide polymorphism in both alleles suggest that the Men113G allele is not recently derived, but we find no evidence in the DNA sequence data for selection maintaining the polymorphism. Interestingly, the alleles differ in both Vmax and Km for the substrate malate. Triglyceride concentration and isocitrate dehydrogenase (IDH) and glucose-6-phosphate dehydrogenase (G6PD) activities are negatively correlated with the in vivo activities of the Men alleles. We examined the causality of the observed correlations using P-element excision-derived knockout alleles of the Men gene and found significant changes in the maximum activities of both IDH and G6PD, but not in triglyceride concentration, suggesting compensatory interactions between MEN, IDH, and G6PD. Additionally, we found significantly higher than expected levels of MEN activity in knockout heterozygotes, which we attribute to transvection effects. The distinct differences in biochemistry and physiology between the naturally occurring alleles and between the engineered alleles suggest the potential for selection on the Men locus. [ABSTRACT FROM AUTHOR]

We studied the potential for response to selection in typical physiological-thermoregulatory traits of mammals such as maximum metabolic rate (MMR), nonshivering thermogenesis (NST) and basal metabolic rate (BMR) on cold-acclimated animals. We used an animal model approach to estimate both narrow-sense heritabilities (h²) and genetic correlations between physiological and growth-related traits. Univariate analyses showed that MMR presented high, significant heritability (h² = 0.69 ± 0.35, asymptotic standard error), suggesting the potential for microevolution in this variable. However, NST and BMR presented low, nonsignificant h², and NST showed large maternal/common environmental/nonadditive effects (c² = 0.34 ± 0.17). Heritabilities were large and significant (h² > 0.5) for all growth-related traits (birth mass, growth rate, weaning mass). The only significant genetic correlations we found between a physiological trait and a growth-related trait was between NST and birth mass (r = -0.74; P < 0.05). Overall, these results suggest that additive genetic variance is present in several bioenergetic traits, and that genetic correlations could be present between those different kinds of traits. [ABSTRACT FROM AUTHOR]

Laboratory selection experiments play a prominent role in understanding organismal adaptation. Although bacteria are not yet commonly used for such experiments, they are well suited for analyses of both the organismic and the genetic basis of adaptation. Bacteria can be maintained in large populations while occupying limited laboratory space, have short generation times, are well characterized physiologically, biochemically, and genetically, and are readily frozen and revived from the freezer. In addition, the genomes of many species are completely sequenced and knowledge of gene function is unparalleled. Here we review general aspects of selection experiments, the history of using selection experiments in combination with thermal biology and genomics, and highlight findings from six lines of Escherichia coli adapted to high temperature (41.5°C), including changes in organismal fitness, physiological performance, gene complement and gene expression. Our results are an example of the powerful insights that can be discovered by combining the tools and analyses of many biological disciplines including genomics, evolutionary biology, genetics, and evolutionary physiology. [ABSTRACT FROM AUTHOR]

Natural selection typically acts on multiple traits simultaneously. Quantitative genetics provides the theory for predicting the response to selection of multiple traits and predicts symmetrical responses to selection (the response to upward selection on both traits is equal to their response to downward selection). In reality, however, the response to simultaneous selection on two traits is often asymmetrical. We provide a physiology-based framework to explain the asymmetrical response to simultaneous selection on two important life history traits: body size and development time. The tobacco hornworm, Manduca sexta, is particularly well suited for such a study, as the physiological control of body size and development time is well known in this species. Three physiological factors control both life history traits in M. sexta: growth rate, the critical weight that measures the timing of the onset of the cessation of juvenile hormone secretion (which initiates the processes leading to pupation) and the time interval between the critical weight and secretion of the molting hormone 20-hydroxyecdysteroid (the interval to cessation of growth, ICG). Asymmetry in the response to simultaneous selection on the two life history traits is due to the different types of selection acting on the three physiological factors. The critical weight and ICG are always under synergistic selection when both focal traits are selected in the same direction and under antagonistic selection when the focal traits are selected in opposite directions. Growth rate follows the opposite pattern. We propose a general model to explain the asymmetric response to simultaneous selection. This model emphasizes the importance of physiological processes in understanding evolutionary responses to selection and the control of complex traits. [ABSTRACT FROM AUTHOR]

The Bayes factor (BF) procedure was applied to examine the additive genetic component of several physiological and vitality variables for newborn pigs. Nine variables were studied: heart rate, arterial oxygen saturation, rectal temperature (all at birth and 60 mm later), birth weight, interval between birth and first teats contact, and interval between birth and first colostrum intake. The available numbers of data ranged from 288 (heart rate at 60 mm) to 839 records (birth weight) from F2 Iberian × Meishan newborn pigs. We compared a model with zero heritability (nonheritable) with the one where the additive genetic background was included. The BF was used to discriminate between both candidate models. Very strong evidence of genetic background was detected for heart rate 60 mm after birth (BF = 48.90), and strong evidence was detected for rectal temperature at birth (BF = 13.82). Posterior modes (means) of heritabilities were 0.29 (0.32) and 0.40 (0.39), respectively. In addition, substantial evidence of absence of genetic background was detected for arterial oxygen saturation at birth. [ABSTRACT FROM AUTHOR]

Reactive oxygen species play a critical role in mediating the oxidative damage that causes senescence in a variety of aerobic organisms, from yeast to mammals. Genetic studies of these organisms have revealed that extended longevity is frequently associated with an increased resistance to stress. However, the relationship between life span and oxidative stress tolerance in plants is poorly understood. We have investigated the responses to oxidative stress in the delayed leaf senescence mutants of Arabidopsis thaliana, ore1, ore3, and ore9. The detached leaves of these mutants exhibit increased tolerance to various types of oxidative stress. The ore1, ore3, and ore9 mutants were also more tolerant to oxidative stress at the level of the whole plant, as determined by measuring physiological and molecular changes associated with oxidative stress. However, the activities of antioxidant enzymes were similar or lower in the mutants, as compared to wild type. These results suggest that the increased resistance to oxidative stress in the ore1, ore3, and ore9 mutants is not due to enhanced activities of these antioxidant enzymes. Taken together, our findings provide genetic evidence that oxidative stress tolerance is linked to control of leaf longevity in plants. [ABSTRACT FROM AUTHOR]

Tocopherols (vitamin E) are lipophilic antioxidants presumed to play a key role in protecting chloroplast membranes and the photosynthetic apparatus from photooxidative damage. Additional nonantioxidant functions of tocopherols have been proposed after the recent finding that the Suc export defective maize (Zea mays) mutant (sxd1) carries a defect in tocopherol cyclase (TC) and thus is devoid of tocopherols. However, the corresponding vitamin E deficient Arabidopsis mutant (vte1) lacks a phenotype analogous to sxd1, suggesting differences in tocopherol function between C4 and C3 plants. Therefore, in this study, the potato (Solanum tuberosum) ortholog of SXD1 was isolated and functionally characterized. StSXD1 encoded a protein with high TC activity in vitro, and chloroplastic localization was demonstrated by transient expression of green fluorescent protein-tagged fusion constructs. RNAi-mediated silencing of StSXD1 in transgenic potato plants resulted in the disruption of TC activity and severe tocopherol deficiency similar to the orthologous sxd1 and vte1 mutants. The nearly complete absence of tocopherols caused a characteristic photoassimilate export-defective phenotype comparable to sxd1, which appeared to be a consequence of vascular-specific callose deposition observed in source leaves. CO2 assimilation rates and photosynthetic gene expression were decreased in source leaves in close correlation with excess sugar accumulation, suggesting a carbohydrate-mediated feedback inhibition rather than a direct impact of tocopherol deficiency on photosynthetic capacity. This conclusion is further supported by an increased photosynthetic capacity of young leaves regardless of decreased tocopherol levels. Our data provide evidence that tocopherol deficiency leads to impaired photoassimilate export from source leaves in both monocot and dicot plant species and suggest significant differences among C3 plants in response to tocopherol reduction. [ABSTRACT FROM AUTHOR]

Examines the effect of feeding spray-dried plasma (SDP) after weaning on the stress response of the pig to a lipopolysaccharide challenge. Quantification of messenger RNA; Potential mechanisms by which SDP may prevent activation of the hypothalamic-pituitary-adrenal axis.

Presents a study that tested a prediction of the ecological model concerning the fitness of hybrids between two young, sympatric species of threespine sticklebacks. Description of the design of the transplant experiment; Growth rate of each species; Average body mass and other morphological traits of the samples.

Presents a study that used sequencing in assessing the genetic diversity of the mitochondrial DNA ND4 gene in the parasite Teladorsagia circumcincta from France and Morocco. Susceptibility and resistance to benzimidazole and nucleotide diversity of the ND4 gene; Correlation to the development of resistance in a worm population and a decrease in genetic variation; Details on the genetic structure.

Presents a study which determined whether the ovine metallothionein 1a-ovine growth hormone (oMt1a-oGH) transgene shows normal Mendelian segregation and whether oMt1a-oGH mice exhibit normal growth in mice even without zinc supplementation. Materials and methods; Relative fitness estimates of transgenic mice in backcross and hemizygous matings; Implications of the results on a growth hormone transgene.

Examines the role of FluG protein in asexual development of Aspergillus nidulans. Genetic screens for dominant suppressors of deltafluG mutation; Identification of components of conidiation signaling pathway acting downstream of FluG; Morphology of dsg1 mutant strains.

The presentation of hypogammaglobulinaemia in young males without a family history of immunodeficiency can pose a diagnostic problem. In the past, the presence of B-cells has suggested a diagnosis of common variable immunodeficiency (CVID), although genotypic analysis has now clarified that individuals with B cells may have mutations in their Btk gene. In order to address the issue of how many male individuals with a clinical diagnosis of CVID do in fact have mutations in the Btk gene, we analysed a group of 24 male patients. Single-strand conformation polymorphism (SSCP) analysis was used to screen the patient cohort for mutations in the Btk gene. Given the size of the Btk gene, the number of patients in the cohort and the amount of available DNA, multiplex PCR reactions were utilized to span the 19 exons and promoter region of the gene. Where abnormal migration patterns were observed with multiplex PCR reactions, in nine of the 24 patients, the individual Btk gene fragments were re-amplified and analysed again by SSCP. Following this analysis, four patients continued to demonstrate abnormal SSCP migration patterns. However, direct sequencing of the relevant Btk gene fragments for these four CVID patients revealed a mutation in only one patient. The mutation was the previously described polymorphism at position 2031 of Btk gene within exon 18. These results indicate that caution should be taken with the application of SSCP analysis to mutation detection. While it has a role to play in screening large patient cohorts, direct sequencing is a necessary adjunct to such analysis. Finally, the clinical diagnosis of CVID in this cohort successfully excluded males with Btk mutations. [ABSTRACT FROM AUTHOR]

cDNA clone (TP5) with significant homology to β-galactosidases has been isolated from a mature tobacco pollen cDNA library by differential screening. The predicted protein of 715 aa shows high levels of homology to plant β-galactosidases expressed during fruit ripening and senescence. Northern analysis shows that the TP5 transcript is expressed exclusively in developing anthers and mature pollen. The transcript is present at very low levels at meiosis and increases dramatically, late in microspore development after mitosis suggesting that the primary role for the protein is during pollen tube growth. β-galactosidase activity, measured by scanning densitometry of histochemically stained tobacco microspores, is first detectable in the early to mid-vacuolate stage, and reaches a peak at microspore mitosis, thereafter decreasing as the microspores reach maturity. Southern analysis indicates that the TP5 gene is present in two copies, probably corresponding to the two ancestral genomes of N. tabacum . [ABSTRACT FROM AUTHOR]