Adhesion molecule expression in acute humoral and acute cellular rejection of renal grafts.
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Biomedical subjects
Publications and source records attributed to G Meduri.
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Monoclonal antibodies have been raised against the porcine LH receptor and have allowed to clone the corresponding messenger RNA from testicular cells. The stricture of the LH receptor has been determined. It shows similarities but also differences with other G protein coupled receptors. Specially a large extracellular domain is specific of that new family of receptors. Variant forms of the LH receptor generated by alternative splicing and lacking transmembrane domain have been isolated. Immunochemical and immunocytochemical studies have been performed. Three different forms of the LH receptor are physiologically expressed: a mature 85kDa transmembrane species, a 68 kDa high mannose containing species corresponding to a precursor which accumulates inside the cells, and truncated 45-48kDa molecular weight species corresponding to the variant messenger RNAs identified during the cloning of the receptor. A novel zonation of the ovary has been described by immunocytochemical studies. Cross hybridization with the LH receptor clone allowed to isolate the related TSH receptor from human thyroid tissue. The human LH and FSH receptor genes have been localized to chromosome 2p21 and the TSH receptor gene to chromosome 14q31. The genes are very large (> 60 kbp) and have introns only within the 5' part encoding the extracellular domain of the receptor. Immunoelectron microscopic studies performed in Leydig cells and in stably transfected L cells have allowed to study intracellular traffic of the LH receptor. The same approach was used to study the transendothelial transfer of hCG in testicular microvasculature.
The entire prostate of a 10 year old boy was cut with a microtome in to 4300 serial slices. The nerves were stained using a monoclonal antibody called anti PS 100. All information was recorded using a computer reconstruction programme. The prostatic nerve supply is very abundant. The nerve fibers of the cranial prostate (central zone) follow a pathway parallel to the anterior surface of the seminal vesicles going towards the caudal prostate. The periurethral zone is widely innervated by nerves arising from the periphery. The caudal prostate also contains many nerve fibers of variable size. We identified many nerve fibers along the anterior surface of the seminal vesicles and surrounding the lateral aspect of the prostatic capsule. They penetrate the capsule and the whole circumference of the caudal prostate. The prostatic capsule is covered by numerous nerve fibers and ganglia, which form a true periprostatic nerve network. The urethra is supplied by numerous thick fibers of more than 30 microns in diameter.
Using 99Tcm-diethylenetriaminepentaacetate (DTPA) an acceptable estimate of glomerular filtration rate (GFR) can be obtained in adult patients by the equation GF (ml/min) = (0.14 x W + 5) x 1000 x k, where W is the 'ideal' body mass (kg) and k the slope determined by two plasma samples. This has been verified in comparison with the results of the Russell (two samples) and Christensen (one sample) methods in 50 patients. The procedure, which does not require determination of the injected dose, could be useful in patients undergoing sequential renal scintigraphy with 99Tcm-DTPA, for example, to check a doubtful value of glomerular filtration obtained by external counting.
Although progesterone and estrogens are essential to maintain human pregnancy after implantation, the localization of their specific receptors in different uterine cell types during pregnancy has not been investigated. We studied uteri (n = 40) obtained during the first 3 months of pregnancy (n = 21) and in late pregnancy (n = 9) as well as from women 5-14 weeks pregnant (n = 10) who had received the antiprogestagen RU 38486 (Roussel-UCLAF) to induce cervical dilation. Frozen tissues were processed for indirect immunocytochemical staining with specific monoclonal antibodies against estrogen receptors (ER; Abbott Laboratories) and progesterone receptors (PR; Li 417). Specific staining for steroid receptors was only detected in the nucleus. In the endometrium, PR staining remained fairly constant throughout pregnancy, whereas ER staining was initially weak and then undetectable. PR was widely expressed in stromal cells and in spiral arterial wall cells, whereas ER was expressed in scattered stromal cells and arterial cells. Both PR and ER were absent from glandular epithelium, contrasting with the secretory activity during the first trimester. Spiral arteries of the endometrium and myometrial smooth muscle cells showed intense PR and moderate ER staining in early pregnancy. The progesterone antagonist RU 38486 (mifepristone), given in early pregnancy at a dose of 200 mg, caused a marked increase in ER staining and a smaller increase in PR staining in stromal cells, whereas the glandular epithelium remained negative for both ER and PR (except for one and two specimens, respectively). We conclude the following. 1) Stromal cells retain PR despite the high progesterone levels during pregnancy, in keeping with the role of progesterone in stromal decidualization. The absence of PR from the secretory glandular epithelium suggests a paracrine link between decidualized stromal cells and epithelial cells. 2) Significant PR down-regulation by progesterone during pregnancy occurs only in epithelial cells of the endometrium. 3) In contrast, the absence or low level of ER staining in the various cell types of the endometrium during gestation concurs with the known effect (down-regulation) of steroid hormones on ER mRNA or protein levels. The increase in ER in human decidua after RU 38486 treatment indicates that the main cause of the low ER levels is progesterone secretion. 4) The intense PR staining in smooth muscle cells of spiral arteries during early pregnancy suggests that progesterone is essential for modulating blood flow during pregnancy.
In order to assess the practical reliability of glomerular filtration rate (GFR) determination with 99mTc-DTPA and plasma sampling, the authors compared the results obtained with 51Cr-EDTA and 99mTc-DTPA in 50 patients using five easily applied methods (two double-plasma-sample methods and three single-plasma-sample methods), and two kits with different compositions. It was observed that: 1) there is no difference between the results obtained with the two different kits. 2) Compared with 51Cr-EDTA the 99mTc-DTPA overestimates the result by about 2 mL/min: precision is slightly lower with 99mTc-DTPA than with 51Cr-EDTA but is sufficient for practical use. 3) The method recommended by the authors on the basis of this experience is the Russell's method with two samples. 4) The simplified methods with one sample give comparable results to the Russell's method for GFR levels between 50 and 115 mL/min, while the results are unsatisfactory below 50 mL/min. 5) Among the single-sample methods, the authors suggest that of Christensen and Groth. 6) A preliminary estimate of GFR (from the serum creatinine level, for instance) is useful for the choice between double-plasma-sample methods and simplified methods. In conclusion, the authors consider that the estimation of GFR with 99mTc-DTPA can be performed efficiently in clinical practice even when operating in absolutely routine conditions.
Monoclonal antibodies have been raised against porcine LH receptor and allowed to clone the corresponding messenger RNA from testicular cells. The structure of the LH receptor have been determined. It shows similarities but also differences to other G protein coupled receptors. In particular a large extracellular domain is specific for that family of receptors. Variants forms of the LH receptor generated by alternative splicing and lacking transmembrane domains have been isolated. Immunochemical and immunocytochemical studies have been performed. Three different forms of the LH receptor are physiologically expressed: a mature 85 kDa transmembrane species, a 68 kDa high mannose containing species corresponding to a precursor which accumulate inside the cells, and truncated 45-48 kDa molecular weight species corresponding to the variant messenger RNAs identified during the cloning of the receptor. A novel zonation of the ovary has been described by immunocytochemical studies. Cross hybridisation with the LH receptor clone allowed to isolate the related human TSH receptor from thyroïds. The human LH and FSH receptor genes have been localized to chromosome 2p21 and the TSH receptor gene to chromosome 14q31. The genes are very large and have introns only within their 5' part corresponding to the extracellular domain of the receptor.
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High endometrial receptivity has been achieved with physiological oestradiol and progesterone replacement cycles in women with ovarian failure. To understand whether different protocols using the oral route or the transdermal route can influence the endometrial maturation and the regulation of sex steroid receptors, we studied 33 women with ovarian failure treated by two commonly used protocols and assessed endometrial receptivity using light microscopy, scanning electron microscopy and immunohistochemistry for oestrogen and progesterone receptors on biopsies taken to include different periods of the luteal phase. The morphology in these patients was similar to that observed in women with normal ovulatory cycles, indicating that the morphological response is not dependent on the type of oestradiol, oral or transdermal, in the replacement cycles as compared to the endogenous oestradiol in the menstrual cycle. The relative distribution of steroid receptors between the epithelium and stroma varies similarly to that observed during the luteal phase of the menstrual cycle. These results confirm the role of progesterone, especially the importance of the number of days of exposure to it, in the disappearance of steroid receptors from endometrial glands. These observations give a better understanding of endometrial receptivity around the time of presumed implantation and confirm clinical results concerning the best timing of oocyte transfer.
Monoclonal anti-LH receptor antibodies were used to study receptor distribution in the various structures of porcine ovary. Primordial and primary follicles were not labeled. Immunoreactivity appeared on the thecal cells of secondary follicles of approximately 100-200 microns in diameter. In large antral follicles and preovulatory follicles granulosa cells were also labeled, whereas two zones were observed in the theca interna. The region close to the lamina basalis (approximately 1/3 of the thecal cells) appeared devoid of LH receptors, whereas a strong labeling was observed on the most external region. Thecal cells were also labeled in atretic follicles. There was no labeling of the oocyte. In cyclic corpora lutea only the most external cells, probably of thecal origin, were stained. The most abundant cells, of granulosa origin, were not labeled. In the interstitium few cells were immunolabeled; they proceeded either from remnants of atretic follicles or existed as groups of isolated cells. Thus both the theca interna of preovulatory follicles and the corpus luteum are heterogenous structures composed of two zones of which only one is directly responsive to LH. No LH immunoactivity could be detected in nontarget organs (liver, lung, thyroid gland, small intestine, fallopian tube, and uterus).
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Monoclonal antibodies have been raised against porcine LH receptor and allowed to clone the corresponding messenger RNA from testicular cells. Cross hybridisation with the LH receptor clone allowed to isolate a clone corresponding to the human TSH receptor from thyroids. The structure of both receptors have been determined. They show similarities but also differences to other G protein coupled receptors. In particular a large extracellular domain is specific of that new family of receptors. Variant forms of the LH receptor lacking transmembrane domains have been isolated. The obtention of monoclonal antibodies against both receptors allowed immunochemical and immunocytochemical studies to be performed. The human LH receptor gene have been localized to chromosome 2p21 and TSH receptor gene to chromosome 14q31. The complete organisation of the human TSH receptor gene has been determined.
We have observed typical cytomegalovirus cytopathology associated with multifocal inflammatory and necrotic lesions of peripheral nerve in biopsy specimens from 4 patients who developed a rapidly progressive, multifocal neuropathy late in the course of human immunodeficiency virus infection. The inflammatory infiltrates, which contained numerous polymorphonuclear cells, were associated with mixed, axonal, and demyelinative lesions of nerve fibers. One of these patients improved on treatment with DHPG (9-[2-hydroxy-l(hydroxymethyl) ethoxymethyl] guanine) and remains stable after 18 months. The other 3 died soon after the onset of the neuropathy. In another patient with acquired immunodeficiency syndrome, who developed a severe, predominantly motor neuropathy of the lower limbs, the nerve biopsy did not reveal cytomegalovirus inclusions, but the neurological deficit improved on treatment with DHPG. The patient died from cachexia 2 months later; numerous cytomegalovirus lesions were found in the spinal cord at the time of postmortem examination. The multifocal necrotic endoneurial nerve lesions with polymorphonuclear cell infiltration we describe may help identify cytomegalovirus neuropathy when characteristic inclusions are not present in the biopsy specimen.
Proacrosin biosynthesis timing during human spermatogenesis has been studied using the monoclonal antibody 4D4 (mAb 4D4). Frozen and paraffin-embedded sections of testicular biopsies were labelled by standard indirect immunofluorescence and avidin-biotin immunoperoxidase procedures. The labelling specificity was checked by immunochemistry assays on unrelated tissues and by western blotting of testis extracts showing that only the 50-55 x 10(3) Mr proacrosin was recognized by mAb 4D4. Proacrosin was first observed in the Golgi region of midpachytene primary spermatocytes. In late pachytene primary spermatocytes, proacrosin was observed in two regions located at opposite nuclear poles. During the subsequent steps of the first meiotic division, the two bodies containing proacrosin were located: (i) on opposite sides of the equatorial plate during metaphase; (ii) along the microtubular spindle during anaphase; and (iii) close to each chromosomal aggregate during telophase. Two bodies containing proacrosin were still observed in interphasic secondary spermatocytes. The single labelled area observed in early spermatids was found to increase considerably in size during spermiogenesis. Anomalies of proacrosin scattering were observed in patients with Golgi complex partitioning failure. These data reveal proacrosin biosynthesis during diploid and haploid phases of human spermatogenesis and the proacrosin partitioning pattern during meiosis.
A method for the purification of epithelial cells from the three anatomical regions of the rat epididymis (corpus, caput and cauda) is described. An enzymic digestion followed by sedimentation of crude cell suspension on discontinuous Percoll gradient yielded quite pure active epithelial cell population as judged by morphological and functional studies. Electron microscopy analysis showed that cells from bands corresponding to densities 1.055 and 1.06 g/ml the gradient preserved a morphology compatible with their epithelial origin and their absorptive and secretory functions. Moreover, they stained positively with anticytokeratin antibody (95-97%) and were negative for antidesmin antibody. They selectively bound L-carnitine through a time-dependent and saturable system and differences in the rate of binding were apparent according to the three anatomical regions of the epididymis.
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