Prenatal Ethanol-induced Effect on Cell Count of Pars Distalis of Pituitary Gland in the Rat Pups.

OBJECTIVE: To determine the effects of prenatal administration of ethanol on cell count in pars distalis of pituitary gland in the rat pups. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Anatomy, College of Physicians and Surgeons Pakistan, Regional Centre, Islamabad, Pakistan, from April 2014 to April 2015. METHODOLOGY: Sixteen female rats (Sprague Dawley) were selected by random sampling method. Rats were mated and divided into control group A and experimental group B. From gestational day 10 to 18, mother rats received intraperitoneal injection of ethanol (Group B) and normal saline (Group A). Mother rats were allowed to complete their gestation and deliver spontaneously. When pups were born, only male pups were selected for the study. They were reared till day five. At 5ᵗʰ; day, pituitary glands were taken out and histological study was done in PAS-OG stain. Cell count was made in unit area (10,000 µ2) of pars distalis of pituitary gland. Student t-test was applied for analysing the data of cell counts in unit area. RESULTS: Mean acidophil count was reduced in experimental group (70.19 ± 11.4) as compared to control group (92.65 ± 8.52, p<0.001). Mean basophil count was reduced in experimental group (27.05 ± 3.9) in comparison to control group (34.03 ± 7.9, p<0.001). Mean chromophobe count was increased (131.95 ± 10.7) in experimental group against the control group (104.62 ± 7.62, p < 0.001). CONCLUSION: Pups exposed to ethanol during gestation, showed significant reduction in acidophil and basophil count while increase in chromophobe cell count in pars distalis of pituitary gland.

Effects of Glucose Administration on Development of Sclera in Chick Embryos.

OBJECTIVE: To determine the effect of glucose administration on the development of sclera in the chick embryo Gallus domesticus. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Anatomy Department, CPSPRegional Centre, Islamabad, from January 2013 to January 2014. METHODOLOGY: The study was carried out in two main groups, control Aand experimental B, which were subdivided into three subgroups comprising 30 eggs each. The group Awas injected with normal saline (0.3 ml) in the egg albumen. The group B was injected with 0.3 ml of 5% w/v solution of glucose equivalent to 15 mg of glucose. Subgroups A1 and B1 were opened on day 10 of incubation. Subgroups A2 and B2 were sacrificed on day 12 of incubation. Eggs from subgroups A3 and B3 were opened on day 15 of incubation. Experimental subgroups were compared with matched control subgroups and quantitative data was analysed statistically. RESULTS: Administration of glucose resulted in changes in thickness of sclera. The mean thickness (µm) of sclera at day 10 of incubation was 43.54 ±2.45 in control subgroup and 43.03 ±5.86 in the experimental subgroup (p=0.673). The mean thickness (µm) of sclera at day 15 of incubation 77.48 ±8.32 in control subgroup and 73.99 ±8.62 in experimental subgroup (p=0.145). The mean number of chondrocytes/unit area of hyaline cartilage of sclera in day 10 was 17.40 ±1.44 control subgroup and 14.57 ±1.87 in the experimental subgroup (p < 0.001). The mean number of chondrocytes/unit area of hyaline cartilage of sclera on day 15 was 10.02 ±0.86 in the control subgroup and 9.54 ±0.59 in the experimental subgroup (p=0.025). There was disrupted ossicular formation indicating adverse effects on the development of bony sclera as well. CONCLUSION: Administration of glucose caused alteration in the histology of sclera in developing chick embryos.

The developmental gross morphology of pancreas in chick embryo after prenatal administration of valproic acid.

OBJECTIVE: To determine the effects of prenatal administration of valproic acid on the developmental gross morphology of pancreas in chick embryo. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Anatomy Department, Regional Centre, College of Physicians and Surgeons, Islamabad, from February 2010 to February 2011. METHODOLOGY: An experimental group-A and control group-B, comprised of 30 eggs each. Freshly laid fertilized chicken eggs of experimental group were injected with valproic acid, incubated and hatched. Eggs of control group underwent sham treatment using normal saline. The chicks were sacrificed on hatching or day 22 of incubation, whichever was earlier. The pancreata of only alive chicks of both groups were dissected out, and evaluated for gross morphology in terms of length and weight by statistically comparing with control ones. Then pancreata were stained with aldehyde fuchsin and orange-G stain to study other obvious histological effects, if any. RESULTS: Chicken embryos exposed to valproic acid in ovo, showed significant decrease in length and weight of pancreata. The mean of length (cm) of pancreata in group-A was 2.208 ± 0.166, and group-B was 2.300 ± 0.102 (p=0.008). The mean of weight (g) of pancreata in group-A was 0.032 ± 0.009, and group-B was 0.048 ± 0.005 (p=0.001). CONCLUSION: Valproic acid exposure showed retarding effect on the gross development of pancreas as depicted by decrease in the length and weight of pancreata.

Effect of passive cigarette smoke on mouse placenta and the role of antioxidants.

OBJECTIVE: To study the effects of passive cigarette smoke on the architecture of mouse placenta and to observe the preventive role of antioxidants if any. METHODS: It was a randomized control trial. Female mice of Balb C strain (51) were mated and grouped as follows: Groups, C: control, S: exposed to smoke and SV: exposed to smoke and given antioxidants (vitamin C, E) and sacrificed at 19 dpc (days post coitus). 14 animals from C, 12 from S and 14 from SV had healthy pregnancies. Their placentae were studied microscopically. The relative thickness of the labyrinthine, spongiotrophoblast-I, spongiotrophoblast-II, and decidual layers were measured. The area of spongiotrophoblast-I was calculated using a computerized software programme. RESULTS: The mean relative thickness and area of the spongiotrophoblast-I in S (14.46 +/- 1.88%, 5.89 +/- 0.87%) was significantly more (p = 0.03, p = 0.035 respectively) as compared to the controls (9.47 +/- 1.31%, 3.95 +/- 0.24%). In SV these values (13.96 +/- 1.2% and 5.74 +/- 0.82%) did not show any significant improvement as compared to S (p = 0.035). The mean relative thickness of the labyrinthine, spongiotrophoblast II, and decidual layer in the control group was 44.49 +/- 2.51%, 19.06 +/- 2.48, 19.44 +/- 0.68 respectively. None of these showed any significant difference from each other in the three groups. CONCLUSION: Cigarette smoke causes a significant disturbance in the architecture of mouse placenta which could not be prevented by antioxidants. Therefore, these effects may be due to other toxic substances present in the smoke rather than free radicals.

Effect of ethanol vapour exposure on survival of chick embryos.

OBJECTIVE: To determine the survival of chick embryos after ethanol vapour exposure by noting the number of dead and alive embryos and comparing with age-matched controls. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Anatomy at the regional centre of College of Physicians and Surgeons, Islamabad, from February 2006 to February 2007. METHODOLOGY: Chicken eggs, placed in an incubator, were exposed to ethanol vapours produced through a specially designed vapour chamber. The experimental group was dissected on day 7, day 10 and day 22 or hatching whichever was earlier and compared with age-matched controls. The proportion of ethanol vapours in the incubator was monitored with a breathalyzer. RESULTS: No statistical difference was seen in the survival of day 7 alcohol-exposed embryos and their age-matched controls. The survival of day 10-control embryos was significantly higher than alcohol exposed group of same age. The embryos exposed to ethanol vapours from day 1 to day 10 and then followed till hatching or day 22, whichever was earlier, had significantly lower survival than age-matched controls. CONCLUSION: In this study, ethanol vapour exposure decreased embryo survival with increasing embryonic age and increased duration of exposure.

Effect of passive tobacco smoking on fertility of female mice.

OBJECTIVE: To determine the adverse effects of passive tobacco smoking on fertility of female mice and the preventive role of antioxidants, if any. STUDY DESIGN: Randomized controlled trial. PLACE AND DURATION OF STUDY: Anatomy Department, CPSP Regional Centre, Islamabad, from February to July 2005. METHODOLOGY: One hundred and seventeen female mice (Balb C) were selected by random sampling. They were mated and grouped as C (n=30) control, S (n=40) exposed to passive smoke in a whole body exposure chamber and SV (n=37) exposed to smoke and given antioxidants (vitamin C, E). At 19 days postcoital they were sacrificed and the number of pregnant animals, fetuses and resorption cases were counted. Histological study of uteri without fetuses was done in H and E stained sections for confirmation of pregnancy. Percentages were calculated and Chi-square test was used to calculate statistical significance. RESULTS: The percentage of pregnancies was 55.00% in S and 80.00% in C (p=0.029). The percentage of animals with more than 11 fetuses was 5.0% in S and 33.30% in C (p=0.001). The percentage of animals with resorption was 31.80% in S and 0.00% in C (p=0.005). These values in SV were 64.86%, 18.90% and 20.80%, which were not significantly different from S (p=0.378, 0.216, 0.390 respectively). Histological study of resorption sites revealed decidual reaction / remnants of the placenta. CONCLUSION: Passive tobacco smoke has adverse effects on fertility of female mice, which were not prevented by antioxidants. Either those were due to other chemicals present in smoke, or the antioxidants were inadequate to neutralize the free radicals.