Cell Journal (Yakhteh)
Volume:2 Issue: 3, 2000

MCP (membrane cofactor protein or CD46) is a complement regulatory proteins (CRPs), which is present in most of body cell membranes (except erythrocytes), and also in most biologycal body fliuds. Due to the important role of this protein in the complement system, specially in fertility, the aim of this study was to determine and isolate this protein in human seminal plasma. 6 semen samples which were considered normal according to W.H.O ceriteria were selected, and then seminal plasma of each sample was separated from sperms and debris by centrifugation. Prostasomes were obtained by ultracentrifugation from seminal plasma then the protein present in prostasome was isolated by FPLC cheromatoghraphy without using any detergent. The presence of MCP was determined by mAb (J4-48) againest CD46 using Dot blot technique. The molecular weight of CD46 was 55KD in first fraction at 6.5ml position by using SDS-PAGE. The presence of MCP in CSDS bound was proved using Western blot technique. Due to usage of such proteins (CD46) in clinical trials, such as infertility, this study shows that it is possible to isolate such proteins without introduction of detergents wich may change physiologycal functions of the protein.

In this study human follicular fluid has been used as an enzyme- rich solution to prevent embryo implantation in mice which resulted in reduction of pregnancy rate. 2-6 month NIH female mice were used to study the effect of interauterine follicular fluid on the fertility rate. 226 plugged female mice were caged in three groups: control, placebo, and experimental groups. Filterated active human follicular fluid was injected on the 4th day of pregnancy transvaginally to the uterine horns of the experimental group. In the pucebo group Ham's F10 medium was injected on the a day and in the same method, and the control group got no injection. After delivery, the number of embryos per mice and pregnancy rate were evaluated. The statistical analysis indicated that: 1) There was a signitificant difference (P<0.05) between the experimental and the other groups, regarding the embryos per mice. ii) In the pergnancy rate, a significant difference (P<0.001) was observed between the experimental and the other groups. No significant difference was observed between the control and the placebo groups in any case. Cunclusion: The transvaginal injection of human fresh follicular flouid in mice at the expected time of implantation may interecept pregnancy and reduce both embryos per mice and the rate of pregnancy. This effect of the pollicular fluid could be due to its negative effects on the endometrial rey in the time of implantation window or per-embryo development or both. The exact mechanism of these and the effective fraction(s) of the folicular fluid in pregnancy interception need mors studies.

Herogeneous proliferative activity of glioma cells (ie non-cycling cells) is on of the main barriers to the therapeutic of radiolabelled IudR. Therefore it is important to evaluate the proliferative heterogeneity on IUdR theoretical because theoretical calculation suggests that this could be the dominant factor in the efficacy of treatment. The uptake of IUdR in solutions of conceteration ranging from 10µM to 100 µM was studied using UVW glioma cell line cultured as monolayer in exponential and plateau phases also spheroids in different range of size (100-200 µm) and 700-1000 µm diameter) and different incubation times, from one to four doubling times by was studied by flow cytometry. The results of the study confirm that there is an inverse relationship between the proportion of cycling cells and spheroid diameter. In monolayer cultures, more than 95% of exponentially growing cells and 62% of plateau phase cells, were labelled with IudR after one doubling time. However, the labelling index in the small spheroids (100-200 µm) was approximately 76% and 28% for large spheroids (700-1000 µm) after one volume doubling time incubation (52 hours). The proporetion of cells that incorporate IUdR in small and large sizes of spheroids inceased with the increasing of the incubation period with IUdR from one to four volume doubling times. Cunclusion: The success of curative targeting strategies is governed by the ability to sterilise all clonogens. Therefore, theraputic regimes must be designed to overcome the limitations imposed by proliterative heterogenety, including the presence of viable tumour cells that are temporarily out of cycle or cycling very slowly.

Listeria monosytogenes is a Gram-positive organism, frequently found in the environment and is responsible for serious food-born diseases such as perinatal infections, septicaemia and meningoencephalitis in humans and animals. Distribution of the CTPA determinant among L. monosytogenes isolated from environmental and clinical samples was investigated using PCR to identify the homologous DNA in 69 isolates, 38% of tested isolates contained the CTPA determinant. DNA homologous to CTPA was not detected in all strains. Our results showed that 90% of clinical and dairy isolates, 85% of environmental isolates and 7% of poultry isolates op L. monocytogenes contained CTPA in chromosomal DNA. It is possible that the increased incidence of CtpA in strains contributes to their association with clinical infection. The occurrence of CtpA was less frequent in poultry isolates and may explain why these strains are not associated with clinical case.

There have been many studies in recent years on the role of nitric oxide (NO) in acute renal failure. In this study, the effects of inhibition or interoduction of No synthase on renal toxicity of gentamicin were investegated. Isolated kidneys of male rats were perfused with Tyrode buffer (T) as the control group, T+L-argrinine as group 2, T+L-NAME as group 3, T+gentamicin (G) as group 4, T+L-arginine + urinary enzymes, lactate dehydrogenace and alkaline phosphatase (markers of cell death) were measured. Enzymes activity in group 2, 3 and 5 were not significantly different from the control group, in contrast groups 4, 6 and 5, 4 were significanthy different. Group 6 showed significant increase and group 5 showed a significant decrease in the enzymes activity as compared with group 4. This study suggests that NO can protect kidney from gentamicin toxicity.

The cell surface glycoconjugate and extracellular matrix may mediate extracellular signals which decide the fate of the cells during tissue interactions. The proper development of cornea needs cellular interaction between surface epithelium, neural crest and developing lens. 50 pregnant rats were chosen and the embryos from pernatal day11-to 20 and postnatal rat from day 1 to 15 were collected. Histochemical staining and lectin histochemistry (PNA, BSA1-B4,S/PNA) were carried out. The presence of D-Gal and Gal/GalNac were confirmed in developing cornea. Sialidase digestion/PNA method did not increase the reaction of cornea to PNA. The statistical analysis showed a significant difference only for neutral saccharide substance. (Mann-Whithey P<0.05). It seems that the pattern of distribution of extracellular matrix and cell surface glycoconjugate in cornea follows a spatiotemporal pattern.

It is clear that the proliferation, migration and differentiation processes of embryonic fetal neural tissue are dependent on special trophic factors. These factors are absent or lessl seen in adult tissue. In this research the effects of Fetal Brain Extract (FBE) on the regenerational processes of lesioned prepheral nerve were investigated. The facial nerve of 16 Wistar rats on the anterior border of the garotid gland was unilaterally cut by a sharp blade. The rats were then divided in to experimental and control groups (n=8). The experimental rats were injected with 0.2 ml of fetal brain extract on 7, 14 and 21 day post operation, while the control rats received normal saline in the same manner. The rats were anesthetized and sacrificed on day 28 post operation and the lesioned region of the facial nerve was sampled (5 mm lenght). The samples were then fixed, processed, embedded in paraffin and cut serially in 7µ thickness and then stained with H & E and picofuchsin. In comparison with the controlgroup a high number of capilary and high amount of connective tissues around the lesiond region existed in the experimental sampels. These results indicate that in the experimental group the trophic factors of fetal extract probably were, retrogradly transported to perikaryons and, in addition to the maintenance of cell body, help them to induce regeneration in the proximal end of nevre fibers. FBE has probably local effects on the connective tissue which are necessary for regeneration.

This investigation was undertaken to examine the influence of thyroid hormone on the development of embryonic CNS during embryogenesis pattern and compare the microscopical and macroscopical structure of the brain in the control and experimental fetuses. 40 Rats, made hypothyroid by chemical thyroidectomy with (PTU). Three weeks later, after T4 and TSH examination, hypothyroid rats mated with euthyroid male rats. Therefore the fetuses of E14, E16 and E20 were collected in two groups as exprimental and control then macroscopical and microscopical studies were carried out. Brain sections with thickness of 10-15mm were pared and stained with H&E and Alcian blue with critical electrolyte concentration with four concentration cations, of Mg. Significant differences were seen in body and brain weight, and Crown Ramp (CR) of embryos in hypothyroid groups. In hypothyroid pregnant rats, it has been reported to cause fetal growth retardation and in this study stillborns, spontaneous abortion and miscarriage were observed in hypothyroid dams. Significant changes in thickness of different layers of pallium and specially in plexiform layer were seen. Thickness of pallium in hypothyroid fetuses in E14 and E16 shows significant difference with P<0.005. It seems that maternal hypothyroidism has an effect on the number of fetuses per dams, and unstability of pregnancy, also the increase of abortion and absorbtion of fetuses may be due to the decrease in the level of serum T4 of the pregnant dams. There is evidence that thyroid hormone in fluences the neuronal development of vertebrate. Thyroid hormone deprivation may delay acquisition of the normal number of cells and abnormal maturation of cerebellar cells.