The estrogen receptor 1 gene affects bone mineral density and osteoporosis treatment efficiency in Slovak postmenopausal women.

BACKGROUND: The study investigated the associations of rs9340799:A > G (XbaI) and rs2234693:T > C (PvuII) polymorphisms in the estrogen receptor 1 gene (ESR1) with femoral neck (BMD-FN) and lumbar spine bone mineral density (BMD-LS), biochemical markers of bone turnover, calcium and phosphate levels, fracture prevalence, and a response to two types of anti-osteoporotic therapy in postmenopausal women from southern Slovakia. METHODS: We analysed 343 postmenopausal Slovak women (62.40 ± 0.46 years). The influence of rs9340799 (AA vs. AG + GG) and rs2234693 (TT vs. TC + CC) genotypes on BMD and biochemical markers was evaluated by covariance analysis adjusted for age and BMI. Binary logistic regression was used to evaluate the genotype effect on fracture prevalence. Pharmacogenetic part of the study included women who received a regular therapy of HT (17ß estradiol with progesterone; 1 mg/day for both; N = 76) or SERMs/raloxifene (60 mg/day; N = 64) during 48 months. The genotype-based BMD change was assessed by variance analysis for repeated measurements. RESULTS: Women with AA genotype of rs9340799 had higher BMD-FN (+ 0.12 ± 0.57 of T-score) and BMD-LS (+ 0.17 ± 0.08 of T-score) in comparison with AG + GG. The rs2234693 polymorphism did not affect any of the monitored parameters. No effect of any ESR1 polymorphisms was found on fracture prevalence. Both types of anti-osteoporotic therapy had a positive effect on BMD improvement in FN and LS sites. Considering the effect of the ESR1 gene within the HT, the subjects with rs9340799/AA genotype showed worse response than those with GG genotype (- 0.26 ± 0.10 of BMD-FN T-score; - 0.35 ± 0.10 of BMD-LS T-score) and also with AG genotype (- 0.22 ± 0.08 of BMD-LS T-score). The rs2234693/TT genotype responded poorer in BMD-LS in comparison with TC (- 0.22 ± 0.08 of T-score) and CC (- 0.35 ± 0.09 of T-score). The effect of the ESR1 gene on raloxifene therapy was reported only in BMD-LS. Subjects with rs9340799/AA genotype had a - 0.30 ± 0.11 of T-score worse response compared to AG genotype. The rs2234693/TT genotype showed - 0.39 ± 0.11 and - 0.46 ± 0.15 lower T-scores in comparison with TC and CC genotypes, respectively. CONCLUSIONS: The rs9340799 polymorphism may contribute to decreased BMD in postmenopausal women from southern Slovakia; however, this is not related to higher fracture prevalence. Concurrently, both polymorphisms affected a response to analysed anti-osteoporotic therapies.

Sperm motility and morphology changes in rats exposed to cadmium and diazinon.

BACKGROUND: Humans are ubiquitously exposed to multiple environmental contaminants. Consequences of combined action on the reproductive system remain unknown. This study aimed to assess single and joint effects of cadmium and diazinon exposure on sperm quality parameters. METHODS: Male adult Wistar rats were randomized into 4 groups of ten animals each. Group A was used as a control, animals from group B were exposed to cadmium (30 mg/L), rats from group C were administered with diazinon (40 mg/L), and rats from group D were exposed simultaneously to cadmium (30 mg/L) and diazinon (40 mg/L) via drinking water for 90 days. Sperm morphology and motility were evaluated using a bright field microscope and a computer-assisted semen analysis. RESULTS: The percentage of motile spermatozoa and morphologically normal sperm was markedly reduced in rats from the group B. Rats from the C group showed an increase in velocity parameters, amplitude of lateral head displacement, decrease in beat-cross frequency, and an increase in abnormal sperm morphology. Simultaneous coexposure to cadmium and diazinon increased distance and velocity parameters, and amplitude of lateral head displacement. Reductions were observed in straightness, linearity, wobble, and beat-cross frequency. The decreased normal sperm morphology rates were related to defects of the sperm tail. CONCLUSIONS: Exposure to cadmium and diazinon at relatively low doses impairs sperm quality and can reduce male fertility. Cadmium and diazinon caused significant changes on sperm morphology with varying effects on motility patterns. These parameters were significantly higher in the group D as compared to the group C. The findings have important implications for reproductive risk assessment of combined exposures to multiple chemicals.

Structural changes in femoral bone microstructure of female rabbits after intramuscular administration of quercetin.

BACKGROUND: Quercetin is one of the best known flavonoids being present in a variety of fruits and vegetables. It has cardioprotective, anticarcinogenic, antioxidant, anti-inflammatory and antiapoptotic properties. Some studies suggest that quercetin has protective effects on bone. However, its influence on qualitative and quantitative histological characteristics of compact bone is still unknown. In our study, 12 clinically healthy five-month-old female rabbits were divided into four groups of three animals each. Quercetin was applied intramuscularly in various concentrations; 10 µg/kg body weight (bw) in the E1 group, 100 µg/kg bw in the E2 group, and 1000 µg/kg bw in the E3 group for 90 days, 3 times per week. Three rabbits without exposure to quercetin served as a control (C) group. Differences in femoral bone microstructure among groups were evaluated. RESULTS: Qualitative histological characteristics of compact bone differed between rabbits from the E1 and E2 groups. Primary vascular longitudinal bone tissue was not found in some areas near the endosteal surface due to increased endocortical bone resorption. In addition, periosteal border of rabbits from the E1 group was composed of a thicker layer of primary vascular longitudinal bone tissue than in the other groups. In all groups of rabbits administered quercetin, a lower density of secondary osteons was observed. Histomorphometrical evaluations showed significantly decreased sizes of the primary osteons' vascular canals in individuals from the E1 and E2 groups. Secondary osteons were significantly smaller in rabbits from the E1, E2, E3 groups when compared to the C group. Cortical bone thickness was significantly increased in females from the E1 and E2 groups. CONCLUSIONS: The results indicate that quercetin has not only a positive dose-response on qualitative and quantitative histological characteristics of the compact bone of female rabbits as it would be expected.

Sex-related variations in bone microstructure of rabbits intramuscularly exposed to patulin.

BACKGROUND: Patulin, a toxic mold metabolite, has been found as natural contaminant of processed fruits, most notably apples, apple juices and other apple-based products. A number of adverse health effects in humans and animals are associated with patulin intoxication. The current study was performed to analyse possible toxic effects of patulin on femoral bone microstructure in adult rabbits in detail. Fourteen clinically healthy four-month-old rabbits of both sexes (6 males and 8 females) were included in the study. Animals from the experimental groups (group E♂, n = 3; group E♀, n = 4) were injected intramuscularly with patulin at dose 10 µg/kg body weight two times a week for 28 days. The dose of patulin was estimated based on the maximum permitted level of patulin for apple products intended for infants and young children. Three males and four females without patulin administration served as controls (groups C♂ and C♀). Cortical bone thickness and qualitative and quantitative histological characteristics of compact bone tissue were investigated. RESULTS: Intramuscular applications of patulin significantly increased the thickness of cortical bone in both sexes of rabbits. In patulin-exposed males, an absence of primary vascular longitudinal bone tissue near the endosteal border was observed, which could be associated with intensive bone remodeling. Femoral diaphyses of females displayed a lower number of secondary osteons in the middle part of the substantia compacta, and occurrence of the osteons near the periosteum. This could indicate alterations in bone turnover. Histomorphometrical evaluations showed significantly increased sizes of the primary osteons' vascular canals (P < 0.05) in males exposed to patulin possibly due to mycotoxin-induced increased levels of testosterone. CONCLUSIONS: This study demonstrates significant impact of intramuscular application of patulin on bone microstructure in adult rabbits. Moreover, we have found that the effects of patulin on qualitative and quantitative histological characteristics of compact bone are sex-dependent.

Effects of subchronic exposure to cadmium and diazinon on testis and epididymis in rats.

The present study aimed to elucidate the structural changes in testis and epididymis of adult rats following subchronic peroral administration of cadmium at 30 mg/L, diazinon at 40 mg/L, cadmium at 30 mg/L, and diazinon at 40 mg/L, respectively. At the end of 90-day experiment, the samples of the testes and epididymis were assayed by qualitative and quantitative histological methods. The testis and epididymis weights increased following exposure to cadmium and simultaneous exposure to cadmium and diazinon. Testicular damage following cadmium and diazinon coexposure was significantly less expressive than in groups with individual administration of these compounds. Cadmium caused a significant thickening of seminiferous epithelium, cellular degeneration, and necrosis. Desquamation of immature germ cells resulted in a significant increase of intraepithelial spaces and reduced tubule volume in all experimental groups. Vascular dilation and congestion were detected in the interstitial tissue. The changes in epididymal histology in the group exposed to cadmium and group exposed simultaneously included a reduction of epithelium, necrotic epithelial cells, vasoconstriction, and interstitial edema together with mononuclear cell infiltration. Results did not indicate a synergistic or any additional effect from the simultaneous administration of both toxicants. Further research is needed to determine the significance and the mechanism of the adverse effects.

Quantitative histological analysis of the mouse testis after the long-term administration of nickel in feed.

In this study, the effects of nickel chloride (NiCl(2)) applied per os on testis histopathology and morphometry of mice were investigated. The metal was applied in pellets at a dose of 10 mg NiCl(2)/kg bw to male mice 4 weeks of age. After 3, 6, 9 and 12 weeks of nickel administration, the relative volume of whole seminiferous tubule, germinal epithelium, tubule lumen, interstitium and blood vessels as well as the diameter of seminiferous tubules were determined in the experimental and corresponding control groups. Microscopic examination of testis showed significant changes in all nickel-exposed groups. The degeneration of germinal epithelium, with released germ cells into the lumen of the tubules, and occurrence of empty spaces in the seminiferous epithelium were found in all experimental groups. The changes in the testes were time-dependent. The relative volume of empty spaces in the seminiferous epithelium significantly increased (P < 0.001) in all experimental groups when compared with the corresponding control. A significant decrease in the relative volume of seminiferous epithelium was observed after 6 and 12 weeks of Ni-exposure. The increased luminization of the tubules was found after 6 (P < 0.001), 9 (P < 0.01) and 12 (P < 0.001) weeks. Interstitial tissue significantly decreased after 6 and 9 weeks of Ni exposure and increased after 12 weeks of Ni intake. The seminiferous tubule diameter significantly (P < 0.001) decreased after 12 weeks. Results of this study report a serious, time-dependent changes in the testes, mainly in the germinal epithelium, after a peroral intake of nickel.