Cell Journal (Yakhteh)
Volume:1 Issue: 4, 2000

Standard semen analysis is routinly used to evaluate the male fertility, however some researchers belive that sperm functional test should also be used with standard semen analysis for more precise evaluation of male fertility. The aim of this study was to consider the relation between different nuclear maturity tests and in vitro fertilization, in order to select the nuclear maturity test for prediction of invitro fertilization. 101 infertil couples were randomly selected form the IVF candidate referring to Isfahan Fertility and Infertility Centre. Semen samples were collected on the day of oocyte recovery. Major portions of the semen were prepared for routine IVF Insemination and the rest was used for following sperm nuclear maturity tests: chromomycin A3 (CMA3), simple acridine orange test- with heat shock, aniline blue and SDS-test. 200 sperm were evaluvated for each test. The result of each test was recorded and were analyzed for its relation to fertilization rate using correlation coefficient, logistic regression analysis, student's t-test and ROC curve. Among these tests only CMA3 and aniline blue showed a signigicant correlation coeficient with in vitro fertilization. How ever, using logistic regression analysis only CMA3 analysis was the independent factor related to in vitro fertilization rate. The results of student's t-test between fertilizing and non-fertilizing groups was also only significant for CMA3. The area under ROC curves suggest that CMA3 is the most spicific and sensitive test among these nuclear maturity tests. This study shows that CMA3 is the most specific and sensitive test among other nuclear maturity tests and independently related to the in vitro fertilization rate. Therefore, CMA3 analysis can be used as a nuclear maturity test along with standard semen analysis for prediction of male infertility.

The contraception has been practiced for over 4000 years and the prevalence of contraceptive use has increased worldwide.The Existing methods of contraception are not perfect and their acceptability is limited by side effects and inconvenience.Thus There is a real need for new methods of contraception to be developed. The effects of the follicular fluid on the pregnancy rate has been studied which can be intoduced as a possible method in interception or implantation control. 201, 3-7 month old, female Sprague Dawley and Albino NMRI rats were obtained from Razi Research Centre and were maintained under standard husbandry conditions with controlled temperature and 12 h light: 12 h dark photoperiod. Four females were caged overnight with males of the same age of proven fertility. Mating was evidenced by the appearance of vaginal plug on the following morning. Plugged females designated day 1 pregnant were caged separately and were devided randomly in three groups: 1) The experimental group which were injected filterated active follicular fluid transvaginally to the uterus on 5th days of pregnancy, 2) The placebo group that were injected Ham's F-10 on the same day, and 3) control group. The number of embryos per animal and percentage of pregnancy were evaluated. The results indicate that statistically significant decrease in the pregnancy rate and the number of embryos per animal in the experimental (follicular fluid) group in comparison with the control and Ham's F-10 groups As implantation is the result of an interaction between the receptive endometrium and the invasive blastocyst; follicular fluid which is an enzym-rich solution may act on either and leads to decrease of the pregnancy rate and number of embryos per rat. The exact mechanisms of these effects need elucidated studies. The results of this study show that the exposure of the endomertium/ or perimplantation embryo to the active follicular fluid might be considered as one of the interceptive methods.

The sexual maturity is a major factor of fertility and reproduction activity. Since no information was availabe about microscopic developement of seminiferous tubules structure including spermatogenic cells, sertoli, PGC (Primordial Germ Cell) and laydig cells, the present detailed study on buffalo testis was undertaken. The present study was made on 30 buffaloes testes, divided into four groups Viz. 1) 3.5-6 months 2) 7-11 months 3) 12-23 months 4) 24-36 months. Tissue samples were processed for paraffin section and stained by Haematoxylin and Eosin stains. The microscopic observation showed that the seminiferous tubules had tubular structure with a define lumen at 3.5 months of age and their epithelium was consisted of abundant spermatogonia, PGC and sertoli cells. The primary and secondary spermatocyte, spermatid and spermatozoa were not seen upto 9 months of age. The coiling and thickness of seminiferous tubules were increased at one year of age. The leydig cells were seen as large clusters in interstitial connective tissue, PGC, sertoli cells, spermatogonia, primary and secondary spermatocyte, spermatid and spermatozoa also were observed in one year age. The development of seminiferous structure were observed continiued with the age. The results showed that, the perfect spermatogenesis and production of spermatozoa is observed in one year age, which it seems similar to cow (10-11 monthsage).

In order to study the morphogenetic role of cell death in the development, a series of comparative studies on the structure of the interdigital membrane of chick (with free digit) and duck (webbed digits) embryos was done. Chick embryos ranging from 6.5 to 10 days of incubation (stag 30-36 H-H) and duck embryos ranging from 8.5 to 12 days of incubation at 12 hour intervals were included in the study. The techniques used in this project were: Vital Staining, Light Microscopy (LM), Transmission Electron Microscopy (TEM). The results of this study showed that interdigital cell death existed both chick and duck embryos, but the intensity of the degenerative change in duck was lower than that of the chick. This relative decrease in the cell death accounts for the survival of the interdigital webs in ducks. The conclusion of this investigation that the interdegital cell death has certain role in separating the digits as well as sculpturing of limb.

The objective of this study was to evaluated effect of Retinoic Acid (RA) on the tibia osteogenesis in the chick embryos. For this purpose 0.75mg RA dissolved in 2ml Dimthylsulfoxide (DMSO) was droped on the limb bud of chick embryos at sixth day of incubation. The osteogenesis of tibia in RA-treated embryos were compared with normal control and DMSO-treated groups (in the same stage) in days 9, 10, 11, 12 and 13 by microscopic and statical analysis. The tibia in all groups were cut, fixed, decalcified, processed in paraffin, serially sectioned and stain with Hematoxyline and Eosin. The volumes of bone formation, cartilage, connective 8 vascular tissue and the volume of the whole of tibia were calculated. Retinoic acid has resulted in an increase in mortality rat and a decreased in the body weigh of the chick embryo. In addition to that, RA has resulted in the reduction of osteogenesis, connective and vascular tissue morphogenesis and resorption of the cartilage and consequently the volume of tibia has reduced. Retinoic acid can retartd Tibia osteogenesis in chick embryos.

This investigation was undertaken to examine chemical nature of glycoconjugate components on the surface of developing rat intestinal cells during gut histogenesis. Rat embryos at days 10-12 of gestation was fixed in B4G and processed for lectinhistochemistry. 5m paraffin sections were incubated with 10-15? g/ml solution of HRP/lecti ns from arachis hypogaea (PNA) and dolichos biflorus (DBA), in 0.1 M PBS, pH 7.2. All sections were developed in substrate medium containing 0.03% DAB and 0.006% H2O2 in tris buffer pH 7.6. Until day 15 of gestation none of two the tested lectin had any reaction with developing intestine. PNA had strong reaction with the submucosal layer of intestine at days 15 and 16. With the exeption and goblet cell the epithelial cells of intestine had strong reaction from day 17 to the time of birth. DBA reacted only with epithelial cells of large intestine two days before birth. The marked changes in the appearance and distribution of the intestinal glycoconjugates during gut morphogenesis support the concept that rapid changes occur in the structure of complex carbohydrates during embryonic and fetal development. The findings also suggest that GalNac and Gal.-GalNac.- containing glycoconjugate may be of functional importance in the regulation of the embryonic intestinal cells during the course of their development.

Nigrothalamocortical tract is one of the main outputs of the basal nuclei, but its role in motor disturbances is vauge, because there is little informations on its connection structure. Some electrophysiological and pharmacological surveys report that none-dopaminergic outputs originate from reticular part of substantia nigra and end at thalamus. In this study the topographical connection of substantia nigra with thalamus was studied using HRP tracer. A tracer, stereotaxically was injected into the MD nucleus of thalamus of 25 rats. The animals were perfused transcardially 48-72 hours later, and the brain tissue was fixed. 40-50 micrometer sections were prepared from diencephalon and midbrain. Following enzymatic reactions, the sections were stained by Tetramethyl-Benzidin method. The light microscopic study showed that there is a high concentration of neurons which were projected to the MD nucleus in the rostro lateral parts of pars reticulata (SNR) and the number of labeled cells decrease in the caudal parts. Other labeled neurons are located at the border of SNC, SNR and VTA, specially close to the passage of III cranial nerve. In general, the size of neurons was small, medium; and multipolar in shape labeled. We didnt see any labeled cell in the substantia nigra opposite to injection site. Our findings shows that none-dopaminergic efferents which originate from substantia nigra nuclei and terminate in MD nucleus. It seems that these fibers not only affect motor mechanisms, but also influence the limbic system and sensory functions.

This study was conducted to evaluate the fibroblast healing of tenotomized Dutch rabbits treated with a low power laser radiation. Male rabbits were divided in a control and exprerimental groups randomly. Achills tendon of animals was cut about 1.5cm above its calcaneal insertion and sutured according to the modified kessler procedure. In the control groups, the tenotomy sites were conventionally treated, while the exprimental groups were treated by low power laser radiation (wave lenghtes 632.8 nm, energy density=10.mj/cm2). Samples were taken in the 14, and 21 days after the operation and processed for EM. The electron microscopic finding demonstrate that the low power laser is capable to enhance the metabolic processe. The fibroblasts had a very well developed rough endeplasmic reticulum in the fibroblast which is also show an increase in collagen deposition.