Neurokinin B Administration Induces Dose Dependent Proliferation of Seminal Vesicles in Adult Rats.

BACKGROUND: Neurokinin B; an endogenous decapeptide, mediates its reproductive physiological actions through gonadotropin releasing hormone. Despite the potential role of Neurokinin B on seminal vesicles, its effects on seminal vesicles in adult male mammals remain elusive. We aimed to investigate the potentials of variable doses of Neurokinin B, its agonist and antagonist on histomorphology and expression of NK3R on seminal vesicles, and secretory activity of seminal vesicles in adult male rats. METHODS: Adult male Sprague Dawley rats (n=10 in each group) were administered intraperitoneally with Neurokinin B in three variable doses: 1 µg, 1 ηg and 10 ρg while, Senktide (Neurokinin B agonist) and SB222200 (Neurokinin B antagonist) in 1 µg doses consecutively for 12 days. After 12 days of peptide treatment, half of the animals (n=05) in each group were sacrificed while remaining half (n=05) were kept for another 12 days without any treatment to investigate treatment reversal. Seminal vesicles were dissected and excised tissue was processed for light microscopy, immunohistochemistry and estimation of seminal fructose levels. RESULTS: Treatment with Neurokinin B and Senktide significantly increased while SB222200 slightly decrease the seminal vesicles weight, epithelial height and seminal fructose levels as compared to control. Light microscopy revealed increased epithelial height and epithelial folding as compared to control in all Neurokinin B and Senktide treated groups while decreased in SB222200. Effects of various doses of Neurokinin B, Senktide and SB222200 on seminal vesicles weight, epithelial height, seminal fructose levels and histomorphology were reversed when rats were maintained without treatments. Immuno-expression of Neurokinin B shows no change in treatment and reversal groups. CONCLUSION: Continuous administration of Neurokinin B and Senktide effect positively while SB222200 have detrimental effects on cellular morphology, epithelial height and seminal fructose levels in seminal vesicles. Effects of peptide treatments depicted a reversal towards control group when rats were kept without any treatment.

Dose-Dependent Degeneration of Leydig Cells Following Kisspeptin-10 Administration: An Ultrastructural Study.

BACKGROUND: The discovery of kisspeptin signaling as a key regulator of gonadotropin- releasing hormone (GnRH) secretion from the hypothalamus enhanced our understanding of the neuroendocrine regulation of mammalian reproduction. Effects of central and peripheral administration of kisspeptin on plasma gonadotropins, testosterone, and spermatogenesis are studied in detail. OBJECTIVE: The present study was conducted to check the ultrastructure of Leydig cells in prepubertal male rats in response to the administration of a range of kisspeptin doses. METHODS: We administered a range of kisspeptin-10 doses (1 µg, 1 ηg, and 10 ρg) intraperitoneally to prepubertal male Sprague-Dawley rats (PND 35) twice daily after every 12 hours. Control rats were injected with physiological saline in parallel. RESULTS: At the end of the treatment, plasma concentrations of testosterone were measured by competitive binding radioimmunoassay, and small pieces of rat testicular tissue were processed for electron microscopy to examine the ultrastructure of Leydig cells. Plasma testosterone concentration was reduced significantly at 1ηg (P<0.05) and 1µg (P<0.01) doses as compared to control. Distinct ultrastructural changes categorized as dilatation of cytoplasmic organelles, irregularly shaped nuclei with nuclear membrane invaginations, reduced nuclear sizes, degeneration, and vacuolation were observed in the kisspeptin-10 treated Leydig cells as compared to control. Quantification of the data showed reduced Leydig cell indices and hyperplasia of the interstitial cells. CONCLUSION: It is concluded that chronic intermittent administration of kisspeptin-10 has a dose-dependent degenerative effect on the plasma testosterone levels and Leydig cells ultrastructure in prepubertal male rats.

To Evaluate and Compare Changes in Baseline Strength of Hairs after Treating them with Deionized Water and Hard Water and its Role in Hair Breakage.

BACKGROUND AND AIM: Interaction of hair with water is common. This study was conducted to compare changes in baseline strength of hair after treating it with hard water and deionized water. MATERIAL AND METHODS: Hardness level of water samples collected from 10 districts of KP, Pakistan was determined, and that with maximum hardness was considered our sample hard water. Hair samples of 70 male individuals, from district with minimum hardness levels, were collected. Each hair sample was divided into three equal parts, and three groups of hair were established, each group containing 70 hairs. Group A was considered control. Group B was treated with deionized water and Group C was treated with hard water. Tensile strength of all three groups was measured using the universal testing machine and compared using paired t-test. RESULTS: The mean age of all 70 participants were 23.87 ± 3. The mean values of tensile strength for hairs of Groups A, B, and C were 255.49, 254.84, and 234.16 with a standard deviation of 57.55, 58.74, and 56.25, respectively. Results were significant in case of hard water (P = 0.001) as compared to deionized water (P = 0.609). CONCLUSION: Hard water decreases strength of hair and thus increases breakage.

Effect of topical application of hard water in weakening of hair in men.

Background: Hard water is thought to play a key role in weakening of hair (not Hair Loss) and breakage especially when travelling is involved. In our community, commonly men do the travelling and complain more about hair problem which is why only young male individuals were included in this study. Materials and Methods: Water samples from different districts of KPK, Pakistan, were collected and their hardness values were estimated to find the water sample of maximum and minimum water hardness in order to know the maximum hardness hair would encounter in KPK, Pakistan. Samples from district Kohat had maximum hardness whereas minimum hardness was estimated in samples of district Peshawar. Water from district Kohat was considered as our sample water for the experimental group of hair. Hair samples were collected from 76 male individuals of district Peshawar, the area with least water hardness among the samples collected. Each hair sample was divided into two halves. One half was considered as experimental group and the other was considered as control group. The experimental group was treated with hard water of district Kohat for 10 minutes on alternate days, for 3 months. In a very similar way the control group was treated with de-ionized water. Tensile strength in term of "Stress" of both the experimental and control groups were measured using the universal testing machine and compared using paired t-test. Results and Conclusions: The standard deviations (SD) for hair treated with hard water and distilled water was 62.05 and 58.13 respectively and the mean values were 238.49 and 255.36 respectively. The results showed that the tensile strength of hair was significantly (p=0.001) reduced in hair treated with hard water as compared to hair treated with de-ionized water.

Human semen quality and sperm DNA damage assessed by comet assay in clinical groups.

BACKGROUND/AIM: About 10%-15% of couples around the world suffer from infertility. Male infertility is responsible directly or indirectly in approximately 60% of cases. A deficiency in semen is the most common cause of male infertility. MATERIALS AND METHODS: The study included 180 male subjects aged 18-50 years with 26 fertile and 154 infertile. The infertile subjects were further subdivided according to the WHO guidelines of semen analysis (2010) into different clinical groups. Sperm DNA damage was estimated using a neutral comet assay. Plasma gonadotropin and testosterone levels were measured using a chemiluminescence assay. RESULTS: The results of the study revealed no significant differences, in semen volume, pH, and liquefaction time between the fertile and all infertile groups. However, sperm concentration, sperm vitality, and sperm motility were significantly lower in all infertile groups as compared to the fertile males. The morphological forms of the sperm and its DNA fragmentation varied significantly between the fertile and infertile males. Reproductive hormone levels were observed to be significantly lower in the infertile than in the fertile males. CONCLUSION: Sperm DNA fragmentation was higher in all of the infertile subjects as compared to the fertile ones. Reproductive hormone levels varied significantly between the infertile patients and the fertile ones.

Insight into the serum kisspeptin levels in infertile males.

BACKGROUND: Regulation of reproduction is now considered to be carried out by the kisspeptin and its receptor, GPR54 or Kiss1r. Mutations of either Kiss1 or Kiss1r in humans and mice result in profound hypogonadotropic hypogonadism. The present study was aimed to determine whether the levels of kisspeptin are associated with male infertility. METHODOLOGY: The study involved 176 male subjects aged 18 - 50 years including 26 fertile and 150 infertile. Infertile subjects were further subdivided according to WHO guidelines of semen analysis into 22 asthenozoospermia, 08 asthenoteratozoospermia, 18 azoospermia, 58 normozoospermia, 06 oligozoospermia, 12 oligoasthenozoospermia and 26 oligoasthenoteratozoospermia. Thorough clinical examinations excluded those suffering from chronic health problems. Serum kisspeptin levels were measured by enzyme immunoassay (EIA) and follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone were estimated by chemiluminescence assay (CLIA). RESULTS: The results of the present study have revealed that kisspeptin levels were significantly lower in all infertile males as compared to the fertile males. Significantly low LH and testosterone levels were observed in all infertile groups as compared to fertile group. FSH levels were significantly lower in normozoospermic and azoospermic as compared to fertile males, while no significant difference was observed between the other infertile and fertile group. CONCLUSION: The study revealed that serum kisspeptin levels were observed significantly lower in the infertile as compared to fertile males, indicating that the kisspeptin might be associated with the fertility problems in males.

The effect of chronic kisspeptin administration on seminal fructose levels in male mice.

The discovery that kisspeptin was critical for normal fertility in all mammalian species including humans, ushered in a new chapter in our understanding of the control of GnRH secretion. Kisspeptin, the product of the KISS1 gene, plays an essential role in the regulation of spermatogenesis acting primarily at the hypothalamic level of the gonadotropic axis. Among the many identified substances in human semen, fructose is becoming increasingly significant. Fructose is synthesized and secreted by the seminal vesicles. Its synthesis is regulated by androgens and it is correlated directly with the levels of testosterone. Dose dependent degeneration of seminal vesicle has been described following intraperitoneal kisspeptin treatment; however, effects of kisspeptin administration on the levels of seminal fructose remain elusive till date. The present study, therefore, addresses the effects of 12-day administration of kisspeptin on seminal fructose levels in male mice. Kisspeptin-10 was administered intraperitoneally at different dosage concentrations (1 µg, 1 ng, and 10 ρg) to adult male mice, twice daily for 12 days. Seminal fructose levels were studied photometrically after 12 days of treatment. At the end of the treatment, seminal fructose levels decreased significantly after all tested doses. Chronic intermittent kisspeptin-10 administration negatively regulates seminal fructose levels in adult male mice.

Kisspeptin-10 induces dose dependent degeneration in prepubertal rat prostate gland.

BACKGROUND: Kisspeptin peptides mediate their actions through the GnRH loop system. How kisspeptins affect prostate gland in prepubertal male mammals remains elusive. METHODS: To address this kisspeptin was administered as subchronic (12 days) twice daily i.p. dose at three different dosage regimens: 10 pg, 1 ng and 1 µg, to prepubertal male Sprague-Dawley rats (PND 35). Control rats were maintained in parallel. At the end of the experiment prostate gland was dissected out and processed for light and electron microscopy. DNA damage was also estimated by DNA ladder assay and DNA fragmentation assay. RESULTS: Prostate weights decreased significantly (P < 0.05) at 1 µg treatment dose of kisspeptin. The epithelial height of secretory acini of prostate decreased at 10 pg (P < 0.05), 1 ng, and 1 µg doses (P < 0.001). Histomorphology and ultrastructure demonstrated, decrease in epithelial cell height, epithelial folding and dilatation of the organelles with kisspeptin treatment. Percent DNA damage to the prostatic tissue was 20.74 ± 2.18, 43.60 ± 2.39, and 58.18 ± 2.59 at 10 pg, 1 ng and 1 µg doses, respectively. CONCLUSION: The study reveals that continuous administration of kisspeptin does not lead to an early maturation but instead severe degeneration of prepubertal prostate gland. Wiley Periodicals, Inc.

Immature rat seminal vesicles show histomorphological and ultrastructural alterations following treatment with kisspeptin-10.

BACKGROUND: Degenerative effects of critical regulators of reproduction, the kisspeptin peptides, on cellular aspects of sexually immature male gonads are known but similar information on accessory sex glands remain elusive. METHODS: Prepubertal laboratory rats were injected kisspeptin-10 at three different dosage concentrations (10 pg, 1 ng and 1 microgram) for a period of continuous 12 days at the rate of two doses per day. Control rats were maintained in parallel. The day following the end of the experimental period, seminal vesicles were removed and processed for light and electron microscopic examination using the standard methods. DNA damage was estimated by DNA ladder assay and DNA fragmentation assay. RESULTS: The results demonstrated cellular degeneration. Epithelial cell height of seminal vesicles decreased significantly at all doses (P < 0.05). Marked decrease in epithelial folds was readily noticeable, while the lumen was dilated. Ultrastructural changes were characterized by dilatation of endoplasmic reticulum and Golgi complex, heterochromatization of nuclei, invagination of nuclear membranes and a decreased number of secretory granules. Percent DNA damage to the seminal vesicle was 19.54 +/- 1.98, 38.06 +/- 2.09 and 58.18 +/- 2.59 at 10 pg, 1 ng and 1 microgram doses respectively. CONCLUSION: The study reveals that continuous administration of kisspeptin does not lead to an early maturation but instead severe degeneration of sexually immature seminal vesicles.

Metabolic syndrome in school children of Dera Ismail Khan, Pakistan.

BACKGROUND: Childhood obesity has increased considerably in many regions of the world including Pakistan. The recent phenomenon of 'nutritional transition' with a westernisation of food so prevalent in developing countries, has caused a significant rise in obesity among population that were unaware of this problem in the recent past. The aim of this study was to find out the frequency of metabolic syndrome and cardiovascular risk factors in obese school children (6-11 years) in Dera Ismail Khan. METHODS: Eighty-six children were included in this study with 61 (70.94%) obese and 25 (29.06%) normal weight children. Obese children comprised of 34 (39.53%) boys and 27 (31.40%) girls. Normal weight children included 15 (17.44%) boys and 10 (11.63%) girls. They were selected among 1.336 children from 8 primary schools of Dera Ismail Khan city. Anthropometric parameters of each subject were recorded, BMI determined and body mass status calculated. Children were categorized by the presence or absence of Obesity. Blood Pressure was also measured. Non-fasting venous blood samples were taken, analysed for lipids; Triglycerides (TG), Cholesterol (TC); Lipoproteins: High and Low Density Lipoprotein-cholesterol (HDL-C, LDL-C) and Plasma Glucose Concentration (PGC). Metabolic syndrome was identified in the presence of > or = 3 of the followings with cut-off values: TG > 170 mg/dl, HDL-C < 35 mg/dl, WC > 71 cm, BP >120/80 mm Hg, PGC > 200 mg/dl. RESULTS: Metabolic syndrome was identified in 22.95% of the obese children. It was 19.67% and 3.27% in obese boys and girls respectively. Metabolic syndrome was not found in normal weight children. Clustering of cardiovascular factors was abundantly present in obese and rare in normal weight children.