Mouse homologues of the human AZF candidate gene RBM are expressed in spermatogonia and spermatids, and map to a Y chromosome deletion interval associated with a high incidence of sperm abnormalities

An RNA-binding motif (RBM) gene family has been identified on the human Y chromosome that maps to the same deletion interval as the 'azoospermia factor' (AZF). We have identified the homologous gene family (Rbm) on the mouse Y with a view to investigating the proposal that this gene family plays a role in spermatogenesis. At least 25 and probably >50 copies of Rbm are present on the mouse Y chromosome short arm located between Sry and the centromere. As in the human, a role in spermatogenesis is indicated by a germ cell-specific pattern of expression in the testis, but there are distinct differences in the pattern of expression between the two species. Mice carrying the deletion Yd1, that maps to the proximal Y short arm, are female due to a position effect resulting in non-expression of Sry ; sex-reversing such mice with an Sry transgene produces males with a high incidence of abnormal sperm, making this the third deletion interval on the mouse Y that affects some aspect of spermatogenesis. Most of the copies of Rbm map to this deletion interval, and the Yd1males have markedly reduced Rbm expression, suggesting that RBM deficiency may be responsible for, or contribute to, the abnormal sperm development. In man, deletion of the functional copies of RBM is associated with meiotic arrest rather than sperm anomalies; however, the different effects of deletion are consistent with the differences in expression between the two species.      

Intertubular capillary changes in kidney allografts: a morphologic investigation on 61 renal specimens.

A total of 61 kidney allograft specimens (51 biopsies and 10 nephrectomies) from 44 patients were investigated. Transplant glomerulopathy (TGP) was found in 38 specimens. Electron microscopy disclosed peculiar intertubular capillary changes (ITCC) in 47 specimens (all those showing TGP and nine without TGP). They were characterized by splitting and multilayering of the basement membranes. TGP and ITCC were graded and found mild (in 23 and 22 specimens), moderate (in 11 and 16 specimens), and severe (in four and nine specimens). Both worsened with time lapse from transplantation. When both were compared in single specimens, their grading was similar or, with a few exceptions, more severe for ITCC. TGP and ITCC were more frequent in biopsies with morphologic evidence of rejection and less frequent in patients treated with Cyclosporine A. Repeated biopsies and nephrectomies were studied in 10 patients and a somewhat more precocious appearance of ITCC than of TGP was found. Earlier and more severe ITCC were found in two patients who had had two transplants as compared to those with one. Morphologic similarities between TGP and ITCC suggest a common pathogenetic mechanism related to endothelial damage. The association to the histological evidence of rejection as well as the apparently protective activity of Cyclosporine A could support the involvement of an immune-mediated mechanism in agreement with what is reported in literature. The constant association of ITCC and TGP and its possible more precocious appearance enforce the diagnostic value of the former, which allows to infer the presence or the subsequent development of TGP even in biopsies where glomeruli are lacking.     

Changes of ovarian microvasculature in hCG stimulated rabbits. A scanning electron microscopic study of corrosion casts

The microvasculature of the ovarian cortex was studied by means of scanning electron microscopy of vascular corrosion casts in estrous and hCG stimulated rabbits in order to establish a better understanding of the structural as well as functional vascular changes which accompany the evolution of the luteofollicular complex. According to the various shape and size seven different morphological types of vascular plexuses corresponding respectively to antral follicles (Types 1-2), atretic follicles (Types 3-4), periovulatory follicles (Type 5), growing pseudopregnant corpora lutea (Type 6) and regressing pseudopregnant corpora lutea (Type 7) were identified. Growing to mature cycling type follicles (Types 1-2) showed a gradual enlargement and proliferation of the theca capillaries. These changes, associated with capillary hyper-permeabilization were observed in ovulatory and post-ovulatory follicles (Types 5), after hCG stimulation. The corpus luteum formation (Types 6) was accompanied by additional capillary dilatation, diffuse angiogenetic sprouts and organization of a more conspicuous venous drainage. The regression of the corpus luteum (Type 7) was characterized by the appearance of avascular areas within the glandular tissue and by regression of vascular dilatation. The atretic follicle (Types 3-4) wall showed large interruptions (avascular areas) and focal invasion of the central cavity by newly formed capillaries randomly arranged. The hCG stimulation did not affect consistently the atretic follicle microvasculature. The present observation shows that both thecal capillary vasodilatation and angiogenetic processes support the gradual increase of ovarian blood flow during follicle growth and corpora lutea formation and that microvascular changes of atretic follicles are possibly related to a type of inflammatory reaction since they seem to be a consequence rather than a primary cause of atresia.     

Variation of the spatial position computed by Roentgen Stereophotogrammetric Analysis (RSA) under non-standard conditions

Roentgen Stereophotogrammetric Analysis (RSA) has been applied to different kinds of research in order to obtain important information in the biomechanics field. Operative requirements change according to the type of investigation and sometimes practical conditions do not always permit one to respect standard specifications. The aim of this paper is to verify the reliability of the system under non-standard conditions, studying the effects of the focus-to-film distance on the determination of 3D co-ordinates of markers. The application of statistical analysis, consisting of two and three way ANalysis Of VAriance (ANOVA), showed that focus-to-film distance is not a significant source of variation, i.e., the system can correctly compute the marker position inside the calibrated space under all conditions of magnification. Since we obtained a fluctuation due to focal distance change of 10(-5) mm (LSD for 95% confidence level), we can assert that this source causes a variation strictly within the limits required by clinical investigations. Thus, this study contributes to improving the flexibility of RSA in clinical applications, making the technical requirements of the radiographic set-up less constraining.     

Recombinant congenic strains of mice from B10.D2 and DBA/2: Their contribution to behavior genetic research and application to audiogenic seizures

Recombinant congenic strains (RCS) represent a series of related strains, each of which carries a small fraction of the genome of one strain (donor strain) on the genetic background of another strain (background strain). Recombinant inbred strains (RIS) are commonly used to identify major gene segregation and linkage and associations between behavior and quantitative trait loci, whereas recombinant congenic strains (RCS) open other complementary leads. The variability in the reactivity of RCS to a trait is thus the expression of few minor-effect genes originating from the donor strain, because the probability that major genes are present in any one RCS is low. Unlike RIS in which minor-effect genes are often masked by major genes, RCS enable the effects of minor genes to be studied. With our method, for a given trait, an estimate can be made of the gene strength distribution as well as an estimate of the minimal number of genes involved having a certain strength.This study was supported by the Centre National de la Recherche Scientifique (URA 1924 and CSEAL-UPS 44, CNRS), Université René-Descartes, Paris V UFR Biomédicale, and the Fondation pour la Recherche Médicale.     

Succinyl derivatives of N-tris (hydroxymethyl) methyl-2-aminoethane sulphonic acid: their effects on the frog neuromuscular junction.

Succinic anhydride (SA) dissolved in Ringer solution buffered with N-tris (hydroxymethyl) methyl-2-aminoethane sulphonic acid (SA-TES solution) potentiates the depolarizing action of acetylcholine (ACh, 10-40 microM) on frog muscle and the tension induced by bath application of this agonist. Applied from one side of a double-barrelled micropipette, SA-TES increases the amplitude of iontophoretically elicited ACh potentials. The potentiation of the effects of ACh by SA-TES does not involve changes in either the activity of the ACh esterase or the input resistance of the muscle membrane. For depolarizations of frog sartorius muscle, dose-response relationships obtained for ACh concentrations from 0.5 to 20 microM indicate that SA-TES increases the apparent affinity of ACh by a factor of 3. SA-TES exerts an "accelerating' effect on the responses elicited by bath-applied ACh; i.e., it increases the rate of depolarization when ACh is added to the bath and the rate of repolarization upon washing out. These effects are particularly marked in preparations treated with neostigmine (3 microM). SA-TES does not potentiate the depolarizing action of agonists which do not contain an ester group. Moreover, the time course of the responses elicited by these compounds is not influenced by SA-TES. SA-TES fails to influence significantly the effects of the neurally released transmitter. Only a 10% increase in the average amplitude of the endplate potentials was observed. SA hydrolyzes in about 30 min at room temperature; however the SA-TES solution retains its activity for several weeks. Succinate is inactive, and so is SA in Ringer buffered with phosphate. The SA-TES solution contains seven succinyl-TES derivatives, which were separated by ion-exchange chromatography and paper chromatography. At concentrations between 1 to 150 microM, these succinyl-TES derivatives affected the ACh-induced contraction of frog rectus abdominus muscle. The most abundant derivative potentiated the action of high doses of ACh, but was inhibitory at lower ones. The other derivatives were mostly inhibitory. These results are discussed in terms of two hypotheses. One postulates the presence of a diffusion barrier formed by groups that bind ACh and are saturated by SA-TES. The other assumes that SA-TES acts directly on the ACh receptor exerting its potentiating effect through a cooperative mechanism.     

Human prostatic-carcinoma cell-line lncap degrades luteinizing-hormone-releasing hormone

Recent evidence suggests a direct antiproliferative effect of LHRH agonists on the prostatic carcinoma cell line LNCaP. In the present study the possible presence of a LHRH degrading activity (LHRH-DA) in soluble fractions of LNCaP cell homogenates has been investigated. The results obtained show that an LHRH-DA is present in the soluble fraction of LNCaP cells with apparent Km and Vmax values of 31.6 mu M and 4.5 pmol/min/mu g protein respectively. The degradation pattern of LHRH is characterized by two major initial degradation products identified as LHRH 1-5 and LHRH 1-6 fragments. The degradation of the tracer [pGlu-H-3]LHRH, used as a substrate, is inhibited by synthetic unlabelled LHRH (IC50 7.9 mu M) and by several LHRH agonists with different kinetics and potencies; the LHRH agonist [DSer-(tBu)(6),Gly(10)-Aza]LHRH was the most potent blocker of LHRH-DA present in LNCaP cells; this enzymatic activity is also inhibited in a dose dependent manner by somatostatin, TRH, bacitracin and dithiothreitol. The LHRH-DA present in the soluble fraction of LNCaP cells does not seem to be modified by the deprivation of steroids from the culture medium, In conclusion, the presence in LNCaP cells of a soluble peptidase able to degrade LHRH might reinforce the possibility that the prostate is a target for the action of LHRH and of LHRH analogs.     

Cross-sectional study of intestinal parasites and Chagas' disease in the Municipality of Novo Air?o, State of Amazonas, Brazil]

A cross-sectional study was performed on the residents of one in every ten households in the town of Novo Air?o, in the northern meso-region of the State of Amazonas, 250 kilometers from Manaus by riverboat. A family cluster sample of 89 dwellings was studied. A stool sample was requested from each of the inhabitants for examination using the Lutz sedimentation and Baermann-Moraes-Coutinho techniques, and blood was taken by venous puncture for Trypanosoma cruzi Elisa antibody testing and immunofluorescence. From a total of 316 stool samples, 87.6% had one or more parasites: Ascaris lumbricoides (35.1%), Entamoeba histolytica (29.1%), Giardia lamblia (17.4%), and other parasites with lower prevalence rates. These results were directly correlated with lack of sanitation and clean water supply. Of the 346 sera examined, 16 (4.6%) were reactive to T. cruzi antibodies, but only three showed a correlation between this result and human contact with wild triatomines, known locally as "piassava lice".