No abstract

The subpopulation of lymphoid cells at the different stages of insulitis in BB rats was determined by immunohistochemical techniques with various monoclonal antibodies, including the recently developed OX41, which distinguishes macrophages from T-lymphocytes, OX19 for pan-T-lymphocytes, W3/25 for both helper T-lymphocytes and macrophages, OX8 for cytotoxic T-lymphocytes and natural killer cells, and OX12 for B-lymphocytes. The major population of infiltrated cells found during the early stages of insulitis appeared to be macrophages. This preceded invasion by a mixed population of cells, including both T- and B-lymphocytes and/or natural killer cells. The preferential infiltration of macrophages during the early stages of insulitis strongly suggested that there might be an initial change in the target beta-cells that precedes their immune destruction, although the amplification of immune response by activated T-lymphocytes and natural killer cells at a later stage seemed to be required for the clinical expression of the disease.

Using an antibody (6313/G2) directed against a specific sequence in the extracellular domain of the type 1 angiotensin II receptor (AT1), we demonstrated the presence of angiotensin II (AII) receptors in human fallopian tube. Immunoperoxidase staining for AT1 receptor showed positive staining in the epithelium of the tubal mucosa. The intensity of staining varied depending upon the hormonal status at the time of salpingectomy, being strongest in the proliferative phase of the ovarian cycle and weakest after menopause. Ligand binding assay confirmed that the AII receptor concentration was highest in the mucosa of fallopian tubes from premenopausal women. Mucosa from the ampullary segment had higher concentrations of AII receptor than the fimbrial and isthmic segments in both premenopausal and postmenopausal women. Displacement studies using specific AII receptor subtype antagonists showed that approximately 60% of the total activity could be displaced by CGP42112B (type 2 specific) and 40% by losartan (AT1 specific). Immunoblotting confirmed that the antibody detected a protein of approximately 60 kDa. Functional studies showed that AII had a stimulatory action on tubal ciliary beat frequency, but had no significant effect on myosalpingseal activity. This effect was achieved at nanomolar concentrations of AII; further increases in the AII concentration were without additional effect. The stimulatory effect of AII was inhibited by the specific AT1 antagonist losartan, whereas the type 2 antagonist, CGP42112B, had no effect. The data demonstrate that AII may play an important role in ovum transport and fertility.

Since Intra-uterine insemination IUI requires the isolation of motile spermatozoaatozoa, advances in andrology research has helped understand the physiology of male germ cell and allowed development of better and more sophisticated techniques to separate functional spermatozoaatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. When compared with other techniques, Density Gradients Centrifugation (DGC) technique allows maximum yield of motile spermatozoa. Several density media like IxaPrep, Nycodenz, SilSelect, PureSpermatozoa and Isolate have been developed to replace Percoll which, was banned in 1996 due to risk of contamination with endotoxins. Semen analysis, according to the revised, WHO (2010) criteria should be carried out prior to processing the sample. Although sophisticated testing—such as DNA fragmentation analysis, oxidative stress analysis and spermatozoa evaluation for genomic, proteomic and metabolic factor are in research phase, it is more than likely that in future these will help in assessing the suitability of the sample in certain cases of male factor or unexplained infertility. The isolation of functional spermatozoaatozoa from highly viscous ejaculates is a challenge that can be performed enzymatically to liquefy the ejaculate. Special care should be taken when processing HIV, Hep B and Hep C, positive samples. Prior to insemination, the processed sample should be tested to ensure the absence of HIV, Hep B and Hep C. There is no agreement between Andrologists as to what should be the minimum motile count for IUI to be successful. Pregnancies have been reported with counts in range of 1 – 10 million motile spermatozoas. Morphology of the processed sample has limited influence on the final outcome. Finally, single insemination 40 h after the hCG injection should be performed using a soft catheter. Normally, 2nd insemination 24 hrs later is only indicated when the follicle has not ruptured at 40 h post hCG injection.