Microsatellite variation of ESR1, ESR2, and AR in Serbian women with primary ovarian insufficiency.

OBJECTIVE: This study aimed to investigate the potential role of microsatellite polymorphisms of the estrogen receptor alpha gene (ESR1) TA repeat, estrogen receptor beta gene (ESR2) CA repeat, and androgen receptor gene (AR) CAG and GGN repeats among Serbian women with primary ovarian insufficiency (POI). These microsatellites have been reported to be associated with POI in different racial/ethnic populations. METHODS: A cohort of 196 POI cases matched with 544 fertile controls was recruited by the Institute for Endocrinology, Diabetes and Metabolic Disorders of Serbia between 2007 and 2010. DNA was extracted from saliva. The four microsatellites were genotyped using a PCR-based assay to determine the repeat lengths. RESULTS: POI patients carried shorter repeat lengths of ESR2 (CA)n than controls (P = 0.034), but the difference was small. ESR1 (TA)n was on the borderline of statistical differences between groups (P = 0.059). AR (CAG)n and (GGN)n showed no association with POI. CONCLUSIONS: We cautiously conclude that microsatellite polymorphisms of gonadal steroid receptor genes might contribute to the genetic basis of POI in Serbian women, but a larger-scale study and family-based studies are warranted to validate our findings even though the sample size in this study is larger than any previously published in this field.

Lack of association between ESR1 gene polymorphisms and premature ovarian failure in Serbian women.

OBJECTIVE: It has previously been reported that estrogen receptor-alpha (ERα) gene (ESR1: estrogen receptor 1) polymorphisms are associated with premature ovarian failure (POF). The aim of this study was to investigate whether these genetic polymorphisms of ESR1 are associated with POF in Serbian women. METHODS: A series of 197 POF cases matched with 547 fertile controls was recruited by the Institute for Endocrinology, Diabetes and Metabolic Disorders of Serbia between 2007 and 2010. Genomic DNA was extracted from saliva using Oragene® DNA sample collection kits. Two single-nucleotide polymorphisms (SNPs), PvuII and XbaI, in ESR1 were genotyped by dynamic allele-specific hybridization. Haplotype analyses were performed with the restriction fragment length polymorphism method. SNP and haplotype effects were analyzed by logistic regression models. RESULTS: No significant difference was found in the distribution of ESR1 PvuII and XbaI polymorphisms or haplotypes between the POF and control groups. CONCLUSION: The two ESR1 SNPs, PvuII and XbaI, are not commonly associated with POF in Serbian women and may not contribute to the genetic basis of the condition.

The divergence between human and baboon globin genes.

Complementary DNA (cDNA) was prepared with viral RNA-directed DNA polymerase from purified baboon globin messenger RNA (mRNA). Homologous and heterologous hybrids between human and baboon mRNAs and cDNAs were compared for extent of hybridisation and thermal stability. Higher mRNA inputs to the hybridizations were required to reach saturation in the heterologous cases. The melting temperature of the heterologous hybrid was 5 degrees C lower than the homologous hybrid. Between these two primates, divergence has occurred in the globin gene to a smaller extent than that possible from third position changes in the coding sequences of the divergence of total DNA. Globin cDNA prepared from baboon will not in general be useful as a probe for human globin mRNA or human globin gene sequences.

Confocal immunofluorescence localization of collagen types I, III, IV, V and VI and their ultrastructural organization in term human fetal membranes.

The distribution of collagen types I, III, IV, V and VI in term human fetal membranes was examined using conventional and confocal indirect immunofluorescence techniques. Collagens I and III were present in most of the layers of fetal membranes except in the trophoblast layer contrary to what has been previously reported. Although collagen IV is considered to be a basement membrane component our study, using monoclonal and polyclonal antibodies, showed its consistent presence in the spongy and reticular layers in high intensity. This was first report on the distribution of type V collagen in the chorion where it was found in the reticular and in the trophoblast layers. Type VI collagen was present mainly in the amnion and the reticular layer. The ultrastructural examination of the extracellular matrix showed that the main fibrous skeleton of the fetal membranes was formed of large banded fibres (Ultrastructurally identical to collagens types I and III) connected together and to the epithelial basement membranes by networks of unbanded filaments (collagen types V, VI and other components). The extensive and continuous networks formed by these collagens may be a major factor responsible for the mechanical integrity of the fetal membranes.

Discordant expression of the G syndrome in monozygotic twins.

We present clinical manifestations of monozygotic male twins with different degrees of expression of the G syndrome. Monozygosity was confirmed using DNA mini-satellite "fingerprint" analysis. The findings in these twins suggest that expression of the G syndrome can be strongly influenced by the prenatal developmental environment.

A colourimetric, microplate assay for the leucotoxin of Pasteurella haemolytica.

Culture supernates of Pasteurella haemolytica, which contain leucotoxin, inhibited the reduction of nitroblue tetrazolium (NBT) by bovine and ovine but not rabbit leucocytes in response to phorbol 12-myristate 13-acetate (PMA). Culture supernates of P. multocida, which contain no leucotoxin, had no inhibitory effect on the response of leucocytes from any species. The inhibition of NBT reduction was assessed visually or spectrophotometrically in the wells of microplates and used as a simple assay for leucotoxin. It was as sensitive as the trypan blue dye-exclusion method and did not require the use of radioisotopes. In addition, sera from P. haemolytica-infected calves inhibited leucotoxin activity in the microplate assay. Thus, inhibition of NBT reduction after stimulation of ruminant leucocytes with PMA can be used as a simple, specific assay for leucotoxin and for anti-leucotoxin antibodies.

Distribution of Pasteurella haemolytica in the respiratory tracts of carrier calves and those subsequently infected experimentally with Dictyocaulus viviparus.

Distribution of Pasteurella haemolytica in the respiratory tracts of calves with no apparent clinical signs of illness and those infected experimentally with Dictyocaulus viviparus was determined so as to define carrier sites for this organism. The calves had been positive by nasopharyngeal swab for either P haemolytica A2 or A1 for at least two months or for over a month, respectively, before slaughter. P haemolytica A1 was acquired following horizontal spread from other infected calves. It was observed post mortem that P haemolytica A1 or A2 resided in the tonsils and retropharyngeal lymph nodes of calves of both groups. In addition to these sites, P haemolytica A1 was also isolated from the right cranial lung lobe of one of the calves from the D viviparus infected group although there was no evidence of pasteurella associated pneumonia. It was concluded that tonsil and retropharyngeal lymph nodes appear to be the most important carrier sites for P haemolytica when compared to other tissues of the bovine respiratory tract.

An atypical case of lymphosarcoma (sporadic bovine leukosis) in a heifer.

An 18-month-old Friesian heifer had several unusual, raised, black, cutaneous plaques, some of which were up to 20 cm in diameter, on its head and neck, limbs, thorax and perineum. There was also generalised lymphadenopathy. A clinical diagnosis of lymphosarcoma (sporadic bovine leukosis) was derived from a fine needle aspiration of a skin lesion. Post mortem and histological examinations confirmed a multicentric lymphosarcoma with widespread infiltration into many of the tissues recognised as predilection sites for this type of tumour. However, in the authors' experience, the presence of tumour masses in the trachea and the right mainstem bronchus was atypical.

An outbreak of strangles in young ponies.

A natural outbreak of strangles occurred in a group of 19 young experimental ponies. The disease was diagnosed in 11 of them within two days of their arrival at Glasgow University veterinary school and five others developed clinical signs within a further four days, a morbidity rate of 84 per cent. All of the affected ponies had typical signs of strangles including dullness, anorexia, pyrexia, regional lymphadenitis, occasionally with rupture of the lymph node, conjunctivitis and a mucopurulent nasal discharge. Nine of the affected ponies were destroyed during the clinical phase of the disease for post mortem studies. The clinical disease in the remaining animals lasted approximately 21 days although one pony had to be destroyed 10 days after the onset of clinical signs because of the development of septic arthritis. All 16 affected animals exhibited peripheral blood neutrophilia and high plasma fibrinogen levels. beta haemolytic streptococci were isolated by nasopharyngeal swabbing from 18 of the 19 ponies. Streptococcus equi was confirmed only in three animals within the first four days of the outbreak. The majority of the other isolates identified to species were S zooepidemicus. beta haemolytic streptococci were still present in six ponies 40 days after they had clinically recovered and were isolated regularly from three ponies which did not develop clinical strangles but remained in contract with affected animals throughout the study.

Collagen gene structure.

The collagens of vertebrates may be divided into three groups according to chain size and whether or not the helical domains are continuous. Present evidence suggests that, at least within one of these groups, similarity between collagens is reflected in the organization of the genes that encode them. Early evidence suggested that collagen genes evolved on the basis of exons which are multiples of a primordial building block of 54 bp, separated by much larger introns. This model of collagen gene evolution is contradicted by the recent discovery of a collagen gene with a single long open reading frame.