Molecular analysis of CAG repeat length of the androgen receptor gene and Y chromosome microdeletions among Jordanian azoospermic infertile males.

Assisted reproductive technology is a common procedure which helps millions of couples who suffer fertility problems worldwide every year. Screening for genetic abnormalities prior to such procedure is very important to prevent the transmission of harmful genetic mutations to future generations. Microdeletions within the azoospermia factor (AZF) region of the Y chromosome and the expansion of the CAG trinucleotides in the androgen receptor (AR) gene are among the susceptible causes of male infertility in different ethnic groups. Such association has never been studied in Jordan. In this study, we compared CAG repeat length between azoospermic infertile and normospermic fertile Jordanian males and we also screened the frequency of Y chromosome microdeletions in the same cohort. The study included 142 nonobstructive azoospermic cases and 145 normospermic controls. Results have shown that the median CAG repeat length in the azoospermic group is 19 ± 2 compared to 19 ± 1.5 (p = .6262) in the control group. Deletions within the Y chromosome AZF region were detected in 7 of 142 cases (4.93%) and no deletions were seen in the control group. The results of this study confirm the importance of the AZF region in normal spermatogenesis, whereas it shows no link between the length of CAG repeats in the AR gene and male azoospermia in Jordanian group examined.

The effect of an extract of Salvadora persica (Meswak, chewing stick) on fertility of male and female mice.

This study investigated the toxic effects of an extract of Meswak from Salvadora persica for 30 days on the reproductive system of the mouse. The results showed that exposure to Meswak extract did not have much effect on female mouse fertility, although it caused a significant decrease in the relative weights of the ovary and an increase in uterine weights. Exposure of male mice to Meswak extract resulted in a 72% reduction in pregnancies in untreated females impregnated by test males. The relative weights of the testes and preputial glands were significantly increased and that of the seminal vesicles was significantly decreased in test males. The results indicate that Meswak has adverse effects on male and female reproductive system and fertility.

Evaluation of the toxic potentials of cypermethrin pesticide on some reproductive and fertility parameters in the male rats.

Adult male Sprague-Dawley rats were exposed to tap water containing 0, 8,571, 17,143, or 34,286 ppm cypermethrin for 12 weeks. Based on water consumption per animal per day the rats received 13.15, 18.93, and 39.66 mg cypermethrin, respectively. Fertility was significantly reduced in male rats ingesting cypermethrin at a concentration of 13.15 and 18.93 mg in that the number of females impregnated by them was significantly reduced. The number of implantation sites was significantly reduced in females mated with males that had ingested cypermethrin at a concentration of 39.66 mg. A significant reduction in the number of viable fetuses was observed in females impregnated by the exposed males at all three doses of cypermethrin. The body weight gain was significantly lower in the treated males. Ingestion of cypermethrin at a concentration of 18.93 or 39.66 mg per day resulted in a significant increase in the weights of testes and seminal vesicles. Preputial gland weights were increased at all three concentrations of cypermethrin. Epididymal and testicular sperm counts as well as daily sperm production were significantly decreased in exposed males. The serum levels of testosterone, follicle-stimulating hormone and luteinizing hormone were significantly reduced in males exposed to 39.66 mg per day. Ingestion of cypermethrin at 18.93 and 39.66 mg/animal/day also resulted in a significant decrease in the perimeter and number of cell layers of the seminiferous tubules. The testes of treated animals were infiltrated with congested blood vessels with marked hemorrhage and a significant accumulation of connective tissue surrounding the seminiferous tubules, which contained a large number of immature spermatids. These results clearly demonstrate the adverse effects of cypermethrin pesticide on fertility and reproduction in male rats.

Effects of extremely low frequency magnetic field on fertility of adult male and female rats.

To investigate the effects of an extremely low-frequency (ELF) magnetic field on their fertility, adult male and female Sprague-Dawley rats were exposed to a 50 Hz sinusoidal magnetic field of approximately 25 microT (rms) for 90 days before they were mated with unexposed counterparts. Exposure to a 50 Hz field reduced male rat fertility. The number of pregnant females was reduced when mated with exposed males, and the number of resorptions increased. The effects of magnetic field on male fertility were shown to be partly reversible, when the same exposed group of males were remated 45 and 90 days after being removed from the fields. Exposure of adult female rats to 50 Hz magnetic fields for 90 days before mating significantly reduced their fertility. The mean numbers of implantations and living fetuses per litter were statistically significantly decreased in the 50 Hz group. These results suggest that low frequency magnetic fields have some adverse effects on fertility of male and female rats.

Assessment of toxic potential of local Jordanian Bacillus thuringiensis strains on Drosophila melanogaster and Culex sp. (Diptera).

AIMS: To assess the toxic potential of different local Jordanian Bacillus thuringiensis isolates on larvae of Drosophila melanogaster and Culex sp. METHODS AND RESULTS: Scanning electron microscopy revealed the presence of spherical, bi-pyramidal, and bi-pyramidal and cuboidal parasporal bodies produced by the toxic isolates. Spherical inclusions dominated. The toxicity of the isolates to the two insects, determined using 24-well plates or vials, indicated that the 50% lethal concentration (LC50) of the bacterial suspension for D. melanogaster and Culex sp. larvae varied from 4.60 to 8.65, and from 5.30 to 6.74, respectively. CONCLUSION: Comparison of the LC50 values of isolate 82 with those of the reference strain B. t. israelensis showed that this isolate has a higher toxicity potential. SIGNIFICANCE AND IMPACT OF THE STUDY: Some local Jordanian B. thuringiensis isolates exhibit toxic potential that could be used to control some important pests, and could replace chemical pesticides.

Effects of long-term exposure to manganese chloride on fertility of male and female mice.

The effect of long-term ingestion of manganese (II) chloride tetrahydrate was investigated on fertility of male and female Swiss mice. Adult male or female mice ingested a solution of manganese chloride along with drinking water at concentrations of 1000, 2000, 4000 and 8000 mg/l for 12 weeks. Fertility was significantly reduced in male mice exposed to manganese chloride solution at a concentration of 8000 mg/l, but not at the other concentrations. There were no treatment-related effects on the number of implantation sites, viable fetuses or the number of resorptions in female rats impregnated by males who had ingested manganese chloride. Fertility was not significantly reduced in female mice exposed to manganese chloride solution at all concentrations used in this study. However, the numbers of implantations and viable fetuses were significantly reduced in females exposed to manganese chloride solution at a concentration of 8000 mg/l. There was no significant effect on the number of resorbed fetuses in females exposed to manganese chloride solution compared to their control counterparts. Absolute body weight was not significantly affected in females exposed to manganese chloride solutions. However, ovarian weight was significantly increased in females exposed to manganese chloride solution at concentrations of 4000 and 8000 mg/l. A significant increase in the uterine weight was also observed at all concentrations used in the study. These results indicate that ingestion of manganese chloride by adult male and female mice causes some adverse effects on fertility and reproduction.

Fetotoxic potentials of Globularia arabica and Globularia alypum (Globulariaceae) in rats.

Adult female Sprague-Dawley rats received by intragastric application, ethanolic extracts of Globularia arabica and Globularia alypum dried leaves. The effect of the extracts was monitored on fertility. The ingestion by female rats of 800 mg/kg of ethanolic extracts of G. arabica and G. alypum, from day 1 to day 6 of pregnancy, did not cause pregnancy failure. However, the ingestion of ethanolic extracts of G. alypum significantly reduced the number of viable fetuses. The number of resorptions was significantly increased in pregnant females administered ethanolic extracts of both G. arabica and G. alypum. The ingestion of 800 mg/kg of ethnologic extracts of G. arabica and G. alypum for 30 consecutive days by adult female rats had no significant effect on the occurrence of pregnancy. However, the ingestion of extracts of both species increased the number of resorptions and only G. alypum extract caused a significant reduction in the number of viable fetuses. These results indicate that ingestion of G. arabica and G. alypum could have some reproductive toxicity in female rats.

Effect of glutathione L-cystein and L-djenkolic acid in the synthesis and mutagenicity of azide metabolite in Bacillus subtilis ATCC 6633 strain.

The Bacillus subtilis ATCC 6633 strain synthesizes a mutagenic metabolite from sodium azide and O-acetylserine. Mutagenicity of azide was decreased in growth media containing 10(-4) M glutathione, L-cysteine or L-djenkolic acid whereas dithiothritol (DTT) added at the same concentration did not reduce the mutagenicity of azide. Likewise, glutathione, L-cysteine, L-djenkolic acid, and DTT were found to have no effect in reducing the mutagenicity of the in vitro produced metabolite using bacterial cell-free extract. These results suggest that O-acetyl-serine sulfhydrylase catalyzes the reaction of azide and O-acetylserine to form a mutagenic metabolite, which is ninhydrin positive and migrates in TLC to an Rf value similar to that of azidoalanine in both acidic and basic solvent systems.

Genetic determinants of active antibiotic-producing soil streptomycetes.

Plasmids or covalently closed circular (CCC)-DNA molecules are abundant in the genus Streptomyces, and have been suggested to be involved in the genetic control of the production of many antibiotics in these organisms. In this study, 21 active antibiotic-producing Streptomyces isolates were screened for their plasmid content by an alkaline lysis method which revealed the presence of a small plasmid DNA in the positive control Streptomyces lividans ATCC 35287, containing pIJ702 plasmid (5.65 kb in size). However, no low molecular weight plasmids were observed in the tested antibiotic-producing Streptomyces strains suggesting that antibiotic production in these strains is likely chromosomally encoded DNA. Treatment of 2 Streptomyces strains with 10 mM ethidium bromide (EB) resulted in the failure to produce aerial mycelia and antibiotic activity.

Sexual maturation and fertility of male and female mice exposed prenatally and postnatally to trivalent and hexavalent chromium compounds.

The reproductive toxicity of trivalent and hexavalent chromium compounds was investigated in male and female mice exposed to 1000 ppm chromium chloride and potassium dichromate via their mother during gestational and lactational periods. Fertility was reduced in male offspring exposed to either trivalent or hexavalent chromium compounds. Body weights and weights of testes, seminal vesicles and preputial glands were reduced in trivalent-exposed male offspring. The exposure of female mice offspring to trivalent and hexavalent chromium compounds delayed sexual maturation. Fertility was reduced in female offspring exposed to either trivalent or hexavalent chromium compounds. The exposure of female mice to hexavalent chromium compound reduced the number of implantations and viable fetuses respectively. Body weight and weights of ovaries and uteri were reduced in trivalent-exposed female offspring. The results indicate that under our experimental conditions, the exposure of male and female mice offspring to either trivalent or hexavalent chromium compounds during gestational and lactational periods impair reproductive functions and fertility in adulthood.

Assessment of aggression, sexual behavior and fertility in adult male rat following long-term ingestion of four industrial metals salts.

1. The effect of long-term ingestion of the industrial metals salts, manganese sulfate, aluminum chloride, lead acetate and copper chloride was investigated on aggression, sexual behavior and fertility in male rat. Adult male rats ingested solutions of these salts along with drinking water at a concentration of 1000 p.p.m. for 12 weeks. 2. Male rat sexual behavior was suppressed after the ingestion of manganese sulfate, aluminum chloride, lead acetate and copper chloride. The ingestion of solutions of these salts markedly prolonged the intromission and ejaculation latencies. Aluminum chloride and copper chloride reduced the copulatory efficiency. 3. Male rat aggression was also abolished after the ingestion of manganese sulfate, aluminum chloride, lead acetate and copper chloride. The ingestion of solutions of these salts markedly suppressed lateralizations, boxing bouts, fight with stud male and ventral presenting postures. 4. Fertility was reduced in male rats ingested with lead acetate. The total number of resorptions was increased in female rats impregnated by males ingested with manganese sulfate and lead acetate. 5. Body, absolute or relative testes, seminal vesicles weights were dropped in adult male rats ingested with manganese sulfate, aluminum chloride, lead acetate and copper chloride. However, the absolute or relative preputial gland weights were not affected. Collectively, these results suggest that the long-term ingestion of manganese sulfate, aluminum chloride, lead acetate and copper chloride would have adverse effects on sexual behavior, territorial aggression, fertility and the reproductive system of the adult male rat.

Reproductive toxicity potentials of Salvia fruticosa (Labiatae) in rats.

The antiimplantation, antifertility and reproductive toxicity potentials after ingestion of aqueous and ethanolic extracts of Salvia fruticosa leaves have been investigated in male and female rats. The ingestion of 200, 400 and 800 mg/kg of aqueous or 400 mg/kg of ethanolic extracts of S. fruticosa from day one to day six of pregnancy by female rats did not cause pregnancy failure. However, the ingestion of an ethanolic extract reduced the number of viable fetuses and increased the number of resorptions in the pregnant rats. The ingestion of aqueous extract (800 mg/kg) or ethanolic extract (400 mg/kg) of S. fruticosa for 30 consecutive days by adult female rats had no effect on the occurrence of pregnancy. However, the ingestion of these extracts reduced both the number of implantations and viable fetuses and increased the number of resorptions in the pregnant females. The ingestion of aqueous extract (800 mg/kg) or ethanolic extract (400 mg/kg) of S. fruticosa for 30 consecutive days by adult male rats had no effect on the number of females impregnated by these males. However, the number of implantations and viable fetuses were reduced in females impregnated by males which ingested either aqueous or ethanolic extracts of S. fruticosa, whereas the number of resorptions was increased in females impregnated by males administered either aqueous or ethanolic extracts of S. fruticosa. On the other hand, the prenatal exposure of male and female rat offspring to 400 mg/kg ethanolic extract of S. fruticosa had no effects on the timing of testicular descent and vaginal opening, respectively. It would appear from these experiments that the ingestion of S. fruticosa may produce adverse effects on the fertility of male and female rats.

Effect of long-term ingestion of chromium compounds on aggression, sex behavior and fertility in adult male rat.

The effects of long-term ingestion of chromium chloride (trivalent compound) and potassium dichromate (hexavalent compound) was investigated on sexual behavior, aggressive behavior and fertility in male rats. Adult male rats were exposed to chromium chloride and potassium dichromate in drinking water at a concentration of 1000 ppm for 12 weeks. The exposure of male rats to chromium chloride and potassium dichromate reduced the number of mounts. The exposure of male rats to potassium dichromate increased the time to ejaculation. On the other hand, the exposure of male rats to chromium chloride and potassium dichromate increased the post ejaculatory interval. The number of animals ejaculating were reduced in chromium chloride and potassium dichromate exposed male rats. The exposure of male rats to chromium chloride and potassium dichromate decreased lateralizations, boxing bouts and fights with stud male. The exposure of male rats to chromium chloride and potassium dichromate had no effect on fertility. Testes, seminal vesicle and preputial gland weights were significantly reduced in chromium chloride- and potassium dichromate-exposed males. In conclusion, the long-term ingestion of chromium chloride and potassium dichromate would have adverse effects on sexual behavior and territorial aggression in adult male rat.

Long-term exposure of male and female mice to trivalent and hexavalent chromium compounds: effect on fertility.

Sexually mature male and female mice at 50 days of age were exposed to trivalent (Chromium chloride) or hexavalent (potassium dichromate) chromium compounds in drinking water for 12 weeks. The effects of the direct chromium exposure on fertility was assessed at day 140 of age. Fertility was significantly reduced in males exposed to the trivalent chromium compound. The number of implantation sites and the number of viable fetuses was significantly reduced in females impregnated by males exposed to the hexavalent chromium compound. The number of resorptions and dead fetuses was increased in females impregnated by males exposed to trivalent and hexavalent chromium compounds. The exposure of female mice to trivalent and hexavalent chromium compounds significantly reduced the number of implantation sites and the number of viable fetuses. The number of females with resorptions was significantly increased in hexavalent chromium exposed females. The number of resorptions was increased in trivalent and hexavalent exposed females. Body, seminal vesicles and preputial gland weights were significantly reduced in males exposed to trivalent and hexavalent chromium, whereas testes weight was significantly increased in males exposed to these compounds. Furthermore, ovarian weight was significantly increased in females exposed to trivalent and hexavalent chromium, whereas uterine weight was significantly decreased in trivalent chromium exposed females. In conclusion, the ingestion of trivalent and hexavalent chromium compounds by adult male and female mice would cause adverse effects on fertility and reproduction.

Developmental studies on Drosophila melanogaster glutathione S-transferase and its induction by oxadiazolone.

Glutathione S-transferase activity toward 1-chloro-2,4-dinitrobenzene was detected in various developmental stages of Drosophila melanogaster. The specific activity of the enzyme was 110, 35, 25 and 15 nmol/min/mg protein in crude extracts prepared from eggs, larvae, pupae and adult stages respectively. The enzymes from larval, pupal and adult stages were purified and compared. Incorporation of the widely used herbicide oxadiazolone at concentrations of 375 and 563 part/million into the culture media caused 4- and 2.5-fold increase in the enzyme activity in pupal and adult stages respectively.

Anterior determinants in embryos of Chironomus samoensis: characterization by rescue bioassay.

Embryos of Chironomus samoensis are programmed, by anterior u.v. irradiation, to form the abnormal body pattern 'double abdomen'. Most double abdomen embryos show a mirror-image duplication of abdominal segments in the absence of cephalic or thoracic segments. Such embryos can be 'rescued', i.e. restored to normal development, by microinjection of cytoplasm or RNA from unirradiated donor embryos. Most of the rescued embryos look completely normal and many of them hatch spontaneously. The rescuing activity decreases from the anterior to the posterior pole in the donor cytoplasm and must be delivered near the anterior pole of the recipient for maximum efficiency. Rescuing activity is present in total RNA extracted from whole, unirradiated embryos. Upon fractionation, the activity is associated with poly(A)+ RNA, with LiCl precipitate depleted of RNA smaller than 250 nucleotides (nt) and with a sucrose gradient fraction depleted of RNA larger than 500 nt. Corresponding fractions of RNA from Xenopus oocytes have no rescuing activity. The activity of Chironomus RNA is sensitive to u.v. irradiation with low fluence affecting less than 2% of the pyrimidine bases. Rescuing activity is present in cytoplasm until the blastoderm stage but disappears earlier from poly(A)+ RNA. Rescuing activity is also present, and localized, in cytoplasm of embryos from two related dipterans, Smittia sp. and Drosophila melanogaster, although the extent of rescue observed in Chironomus decreases with the phylogenetic distance between donor and recipient. The results of these and previous experiments indicate that dipteran embryos contain localized RNP particles acting as anterior determinants. In Chironomus, the activity of these particles seems to depend on the integrity of polyadenylated RNA of about 250 to 500 nt length.