Developmental toxicity of bromoacetamide via the thyroid hormone receptors-mediated disruption of thyroid hormone homeostasis in zebrafish embryos.

Bromoacetamide (BAcAm) is a nitrogenous disinfection by-product. We previously found that BAcAm induced developmental toxicity in zebrafish embryos, but the underlying mechanisms remain to be elucidated. Since thyroid hormones (THs) homeostasis is crucial to development, we hypothesized that disruption of THs homeostasis may play a role in the developmental toxicity of BAcAm. In this study, we found BAcAm exposure significantly increased mortality and malformation rate, decreased hatching rate and body length, inhibited the locomotor capacity in zebrafish embryos. BAcAm elevated TSH, T3 and T4 levels, down-regulated T3/T4 ratios, and up-regulated mRNA expression changes of THs related genes (trh, tsh, tg, nis, tpo, dio1, dio2, ugt1ab，klf9 and rho), but down-regulated mRNA expression changes of TH receptors (tr α and tr ß). Up-regulated tr α and tr ß mRNAs by rescue treatment confirmed that both tr α and tr ß were involved in the developmental toxicity of BAcAm. In conclusion, our study indicates disruption of THs homeostasis via the thyroid hormone receptors was responsible for the developmental toxicity of BAcAm.

Drinking water elements constituent profiles and health risk assessment in Wuxi, China.

Water elements pollution has attracted public attention globally. Wuxi is located in East China, and its water source, Taihu Lake, has been severely polluted since 2007. Studies of elemental pollution profiles have yet to be conducted in this area. In this study, 56 water samples were collected in 2018, and 33 elements were determined using inductively coupled plasma-mass spectrometry (ICP-MS). The results showed that the levels of 33 elements ranged from 1.35 × 10-3 µg/L(Tl) to 101 mg/L(Ca), with Sr, Al, Fe, B, Ti, Ba, and Zn levels being relatively higher. A comprehensive literature review showed spatial distribution of conspicuous elements in drinking water worldwide. Meanwhile, Monte Carlo simulations were applied to evaluate exposure health risks. The total hazard index(HI) for 14 non-carcinogens and the average incremental lifetime cancer risk (ILCR) of As and Pb exposure through drinking water were found acceptable. Sensitivity analyses suggested that Sb and As in the drinking water represent an increasing risk to human health. The results of this study provide key data on local metal pollution characteristics, help identify potential risk factors, and contribute to the development of effective environmental management policies for Taihu Lake.

A Mechanistically Inspired Halenium Ion Initiated Spiroketalization: Entry to Mono- and Dibromospiroketals.

Employing halenium affinity (HalA) as a guiding tool, the weak nucleophilic character of alkyl ketones was modulated by the templating effect of a tethered 2-tetrahydropyranyl(THP)-protected alcohol towards realizing a bromenium ion initiated spiroketalization cascade. Addition of ethanol aided an early termination of the cascade by scavenging the THP group after the halofunctionalization stage, furnishing monobromospiroketals. Alternatively, exclusion of ethanol from the reaction mixture biased the transient oxocarbenium towards α-deprotonation that precedes a second bromofunctionalization event thus, furnishing dibrominated spiroketals. The regio- and stereoselectivity exploited in the current methodology provides a novel and rapid access to the dibrominated spiroketal motifs exhibited by several natural products.

Developmental toxicity of disinfection by-product monohaloacetamides in embryo-larval stage of zebrafish.

As an emerging class of nitrogenous disinfection by-products (N-DBPs), haloacetamides (HAcAms) have been widely detected in drinking water. Limited toxicity studies have shown an inconsistent toxicity of monoHAcAms, including CAcAm, BAcAm and IAcAm. In this study, the developmental toxicity of monoHAcAms was evaluated in embryo-larval stage of zebrafish. Embryos were exposed to one concentration of 2.50, 5.00, 10.0, 20.0, 40.0 and 80.0 mg/L monoHAcAms from 4 h post-fertilization (hpf) to 120 hpf. Multiple endpoints, including hatching rate, morphological abnormalities, mortality as well as locomotor behavior were assessed at specified stages (24, 48, 72, 96 and 120 hpf). Results showed that 80 mg/L CAcAm and 40 mg/L BAcAm significantly decreased the hatching rate, IAcAm decreased the hatching rate and delayed the hatching process in a concentration-dependent manner with an EC50 of 16.37 mg/L at 72 hpf. The frequency and severity order of morphological abnormalities increased with the raised exposure concentrations and prolonged exposure time, and the corresponding EC50 at 96 hpf were 21.10, 9.77 and 16.60 mg/L for CAcAm, BAcAm and IAcAm, respectively. MonoHAcAms exposure resulted in a time- and dose-dependent response in mortality and the calculated LC50 at 72 hpf were 38.44, 17.74 and 28.82 mg/L for CAcAm, BAcAm and IAcAm, respectively. Based on EC50 for morphological abnormalities and LC50, a toxicity rank order of BAcAm > IAcAm > CAcAm was observed. Different degrees of hyperactivity and hypoactivity were observed from locomotor behavior analysis in larvae from ≤10.0 mg/L monoHAcAms exposure groups. The light-dark periodic change was disappeared in larvae of 10.0 mg/L BAcAm exposure group. In summary, our study showed that monoHAcAms were developmentally toxic to zebrafish even at very low concentrations and BAcAm exerted higher toxicity than IAcAm and CAcAm. These results will further our understanding of the toxicity of HAcAms and its potential toxicological impact on human and ecological environment.

Association of semenogelin (SEMG) gene variants in idiopathic male infertility in Chinese-Han population.

Infertility is known to occur frequently worldwide, and the incidence is continuing to rise in China. It is known that semenogelin (SEMG) protein secreted by human seminal vesicles plays an important role in male reproductive system function. However, an association between alterations in SEMG gene functions and idiopathic male infertility occurrence in Chinese-Han population has not been examined. The aim of this study was thus to investigate the inherent relationship between SEMG gene alterations and idiopathic male infertility using a method of variant genotyping selection and semen quality analysis. A population of 484 males with clinically diagnosed idiopathic male infertility and 246 fertile controls were selected after signing consent forms. Results demonstrated a significantly increased frequency of idiopathic infertility with abnormal semen parameters such as semen volume, sperm concentration, sperm number per ejaculate, and sperm motility in variants carrying the rs2301366 TA genotype. Combined association analysis from target single-nucleotide polymorphisms (SNPs) was selected from the genotype database of unrelated Chinese-Han in Beijing individuals from the Hap Map. SNP array analysis in blood samples in each group was carried out by TaqMan Universal PCR Master Mix and TaqMan SNP Genotyping Assays. In addition, the interaction between SEMG SNPs and binding protein epididymal protease inhibitor (EPPIN) SNPs was determined. Our findings demonstrated that the presence of SEMG SNPs and EPPIN SNPs increased the frequency of idiopathic male infertility in Chinese-Han population. It is proposed that measurement of SEMG SNPs and EPPIN SNPs in carriers may thus be utilized to identify idiopathic male infertility.

Maternal air pollution exposure and preterm birth in Wuxi, China: Effect modification by maternal age.

BACKGROUND: Numerous studies have investigated prenatal air pollution and shown that air pollutants have adverse effect on birth outcomes. However, which trimester was the most sensitive and whether the effect was related to maternal age is still ambiguous. OBJECTIVES: This study aims to explore the association between maternal air pollution exposure during pregnancy and preterm birth, and if this relationship is modified by maternal age. METHODS: In this retrospective cohort study, we examine the causal relationship of prenatal exposure to air pollutants including particulate matters, which are less than 10 µm (PM10), and ozone (O3), which is one of the gaseous pollutants, on preterm birth by gestational age. A total of 6693 pregnant women were recruited from Wuxi Maternal and Child Health Care Hospital. The participants were dichotomized into child-bearing age group (< 35 years old) and advanced age group (> = 35 years old) in order to analyze the effect modification by maternal age. Logistic and linear regression models were performed to assess the risk for preterm birth (gestational age < 37 weeks) caused by prenatal air pollution exposure. RESULTS: With adjustment for covariates, the highest level of PM10 exposure significantly increased the risk of preterm birth by 1.42-fold (95% CI: 1.10, 1.85) compared those with the lowest level in the second trimester. Trimester-specific PM10 exposure was positively associated with gestational age, whereas O3 exposure was associated with gestational age in the early pregnancy. When stratified by maternal age, PM10 exposure was significantly associated with an increased risk of preterm birth only in the advanced age group during pregnancy (OR:2.15, 95% CI: 1.13, 4.07). The results suggested that PM10 exposure associated with preterm birth was modified by advanced maternal age (OR interaction = 2.00, 95% CI: 1.02, 3.91, Pinteraction = 0.032). CONCLUSION: Prenatal air pollution exposure would increase risk of preterm birth and reduced gestational age. Thus, more attention should be paid to the effects of ambient air pollution exposure on preterm birth especially in pregnant women with advanced maternal age.

A Near-Infrared Photoswitchable Protein-Fluorophore Tag for No-Wash Live Cell Imaging.

FR-1V, a fluorene-based aldehydic chromophore, binds its target protein as an imine to yield a highly bathochromic pigment, CF-2, a prototypic protein-dye tagging system whose NIR emission can be spatiotemporally switched ON by rapid UV-light activation. This is achieved through photoisomerization of the imine and its subsequent protonation. We demonstrate a no-wash protocol for live cell imaging of subcellular compartments in a variety of mammalian cell lines with minimal fluorescence background.

[The association study between Eppin gene polymorphisms and idiopathic male infertility].

OBJECTIVE: To explore the correlation between four tagSNPs of Eppin gene (rs6124715, rs2231829, rs2227290 and rs11594) and the risk of idiopathic male infertility in the Chinese Han population. METHODS: A total of 473 confirmed infertile patients (from March 2005 to March 2007) and 198 fertile male controls (March 2005 to February 2009) were selected from two hospitals in Nanjing. All the subjects were Han Chinese and came from Nanjing or its surrounding areas. 5 ml peripheral blood was drawn from each subject with informed consent. Four tagSNPs (rs6124715, rs2231829, rs2227290 and rs11594) in Eppin gene were analyzed by the PCR-restriction fragment length polymorphisms (PCR-RFLP) method. The serum testosterone level was evaluated by radioimmunoassay (RIA). RESULTS: The genotype frequencies of AA,AC and CC at rs11594 were 76.3% (361/473), 20.1% (95/473) and 3.6% (17/473) respectively in the case group, while the frequencies in the control group were 75.3% (149/198), 24.2% (48/198), 0.5% (1/198) respectively, the differences were statistically significant (χ² = 7.73, P = 0.021), the CC genotype carriers had an increased risk of male infertility (OR = 7.02, 95% CI:0.93-53.19). In the combined genotype analysis, the haplotype CTGA carriers has significantly lower onset risk (OR = 0.18, 95% CI:0.06-0.53). In the two groups, the frequencies and the risk of male infertility were no statistically significant in rs6124715, rs2231829 and rs2227290 genotype. CONCLUSIONS: The Eppin gene polymorphisms were correlated to the susceptibility to idiopathic male infertility. Among them, CC genotype at rs11594 could increase the risk of idiopathic male infertility.

Long-Term Atmosphere Surveillance (2016-2021) of PM2.5-bound Polycyclic Aromatic Hydrocarbons and Health Risk Assessment in Yangtze River Delta, China.

Long-term atmospheric quality monitoring of fine particulate matter (PM2.5) and PM2.5-bound polycyclic aromatic hydrocarbons (PAHs) was performed in Wuxi from 2016 to 2021. In total, 504 atmospheric PM2.5 samples were collected, and PM2.5-bound 16 PAHs were detected. The PM2.5 and ∑PAHs level decreased annually from 2016 to 2021, from 64.3 to 34.0 µg/m3 and 5.27 to 4.22 ng/m3, respectively. The benzo[a]pyrene (BaP) levels of 42% of the monitoring days in 2017 exceeded the recommended European Union (EU) health-based standard of 1 ng/m3. Five- and six-ring PAHs were found, including benz[a]anthracene, benzo[k]fluoranthene (Bkf), BaP, and benzo[g,h,i]perylene, which were the dominant components (indicating a prominent petroleum, biomass, and coal combustion contribution) using molecular diagnostic ratios and positive matrix factorization analysis. Moreover, PM2.5 and PAHs were significantly negatively associated with local precipitation over a period of six years. Statistically significant temporal and spatial distribution differences of PM2.5, and ∑PAHs were also found. The toxicity equivalent quotient (TEQ) of total PAHs was 0.70, and the TEQ of BaP (0.178) was the highest, followed by that of Bkf (0.090), dibenz[a,h]anthracene (Dah) (0.048), and indeno[1,2,3-cd]pyrene (0.034). The medians of the incremental lifetime cancer risk for long-term exposure to PAHs were 2.74E-8, 1.98E-8, and 1.71E-7 for children, teenagers, and adults, respectively, indicating that the carcinogenic risk of PAHs pollution in air was acceptable to local residents in this area. Sensitivity analysis revealed that BaP, Bkf, and Dah significantly contributed to carcinogenic toxicity. This research provides comprehensive statistics on the local air persistent organic pollutants profile, helps to identify the principal pollution source and compounds, and contributes to the prevention of regional air pollution. Supplementary Information: The online version contains supplementary material available at 10.1007/s12403-023-00572-x.

Quantitative determination of bromochloroacetamide in mice urine by gas chromatography combined with salting-out assisted dispersive liquid-liquid microextraction.

Bromochloroacetamide (BCAcAm) is the main haloacetamide (HAcAm) detected in drinking water in different regions and exhibits strong cytotoxicity and genotoxicity. However, there is no appropriate method for detecting BCAcAm in urine or other biological samples, and thus, the internal exposure level in the population cannot be accurately assessed. In this study, a gas chromatography-electron capture detector (GC-ECD) was combined with salting-out assisted dispersive liquid-liquid microextraction (SA-DLLME) to develop a rapid and robust method for BCAcAm detection in urine of mice continuously exposed to BCAcAm. The factors influencing the pre-treatment procedure, including the type and volume of extraction and disperser solvents, extraction and standing time, and the amount of salt, were evaluated systematically. Under the optimised conditions, the analyte achieved good linearity in the spiked concentration range of 1.00-400.00 µg L-1, and the correlation coefficient was higher than 0.999. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.17 µg L-1 and 0.50 µg L-1, respectively. The recoveries ranged from 84.20% to 92.17%. The detection of BCAcAm at three different calibration levels using this method afforded an intra-day precision of 1.95-4.29%, while the inter-day precision range was 5.54-9.82% (n = 6). This method has been successfully applied to monitor the concentration of BCAcAm in mouse urine in toxicity experiments and can provide technical support for assessing human internal exposure levels and health risks in later studies.

Occurrence and multi-pathway health risk assessment of trihalomethanes in drinking water of Wuxi, China.

Chlorination is widely used to disinfect drinking water to keep humans safe from microorganisms. During chlorination, chlorine and its compounds react with contaminants to form disinfection by-products (DBPs). Toxicological and epidemiological studies have demonstrated that trihalomethanes (THMs) are the most widely investigated DBPs in drinking water, and their exposure has been associated with some adverse health effects. However, studies about risk characteristics in this field are limited. We estimated the health risks of THMs exposure in drinking water through multi-pathways, and systematically analyzed the factors influencing health risks of THMs in Wuxi, China. A total of 488 drinking water samples were collected and analyzed for THMs from four water treatment utilities from 2008 to 2016 in Wuxi. And water exposure parameters were obtained from 602 participants by structured questionnaires. The median concentration of THMs ranged from 6.71 µg/L to 9.18 µg/L. The cumulative cancer risk of THMs exposure through multi-pathways was 1.26 × 10-4, and CHBr2Cl made the largest contribution to the total cancer risk (48.25%). The non-cancer risk of THMs exposure was 2.02 × 10-1. Health risks of the exposure to THMs in drinking water in summer were significantly higher than that in winter (P = 0.0003 for cancer risk, and P = 5.95 × 10-7 for non-cancer risk). In our study, the average individual disability-adjusted life years (DALYs) lost was 1.27 × 10-4 per person-year (ppy). This study attempted to use DALYs for risk assessment of THMs, which will provide useful information for risk comparison and prioritization of hazards in drinking water. This suggested that potential higher risk might exist, and possible measures could be considered to decrease the health risks.

Urinary trihalomethane concentrations and liver function indicators: a cross-sectional study in China.

While it is known that exposure to disinfection by-products (DBPs), including trihalomethanes (THMs), impairs liver function, few epidemiological studies have explored this association. Here, we determined the concentrations of four urinary trihalomethanes (chloroform [TCM], and three Br-THMs, bromodichloromethane [BDCM], dibromochloromethane [DBCM], and bromoform [TBM]), and nine serum liver function indicators in 182 adults ≥ 18 years of age, examined at a medical examination center in Wuxi, China, in 2020 and 2021. Generalized linear model analysis revealed positive associations between urinary DBCM and alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), total protein (TP), and albumin (ALB). Urinary Br-THMs and total THMs (TTHMs) were positively associated with ALT, AST, TBIL, indirect bilirubin (IBIL), TP, and ALB (all P < 0.05). Urinary THMs were not associated with alkaline phosphatase (ALP) or glutamine transaminase (GGT) (all P > 0.05). Generalized additive model-based penalized regression splines were used to confirm these associations. In conclusion, THM exposure was associated with altered serum biomarkers of liver function.

Association between Urinary BPA Substitutes and Precocious Puberty among Girls: A Single-Exposure and Mixed Exposure Approach from a Chinese Case-Control Study.

There is an argument that BPA substitutes may have the same or more deleterious health effects as BPA due to their structural similarity. This study explored the association between urinary BPA substitutes and precocious puberty among girls by including 120 girls with precocious puberty (cases) aged 2-10 years enrolled at Nanjing Children's Hospital Department of Endocrinology in China between April 2021 to September 2021 and 145 healthy girls (controls) recruited from a primary school. Logistic regression was used to evaluate the effect of single exposures, and Bayesian kernel machine regression (BKMR) and quantile-based g-computation were used for the mixed effect. In the multivariate logistic regression, BPS (bisphenol S), TBBPA (tetrabromobisphenol A), and BPFL (bisphenol-FL) were significantly associated with increased risk of precocious puberty (odds ratio (OR) = 1.75, 95% confidence interval (CI): 1.13, 2.76, p = 0.014), (OR = 1.46, CI: 1.06, 2.05; p = 0.023), and (OR = 1.47, CI: 1.01, 2.18; p = 0.047), respectively. The BMKR and quantile-based g-computation models revealed consistent associations for single exposures and there was insufficient evidence for the associations of the mixed exposure of bisphenols with precocious puberty. In conclusion, BPA substitutes such as BPS, TBBPA, and BPFL may be associated with an increased risk of precocious puberty in girls.

Association of CLOCK gene variants with semen quality in idiopathic infertile Han-Chinese males.

Recent experimental animal studies suggested that the circadian locomotor output cycles kaput protein gene (CLOCK) may play an important role in male reproduction. So far, such data for humans are not available. This study used single-nucleotide polymorphisms (SNP) to examine the association between CLOCK and semen quality in a human population with idiopathic infertility. Three-variant genotyping of CLOCK and semen analysis were performed in 478 men with idiopathic infertility by SNP genotyping assays and computer-aided sperm analysis. Subjects carrying a C allele at rs3749474 (CC and TC) presented significantly lower semen volume (P=<0.001 and 0.001, respectively) compared with the TT genotype. Subjects carrying the rs3749474 CC genotype had significantly lower sperm number per ejaculate (P=0.026) and sperm motility (P=0.021) than TT genotype carriers. rs1801260 TC genotype carriers had significantly lower sperm motility compared with the TT genotype (P=0.028). For the rs3817444 genotypes, CA and AA genotype carriers presented significantly lower semen volume compared with the CC genotype (P=0.022 and 0.001, respectively). The findings suggest, as far as is known for the first time, an association between CLOCK genetic variants and altered semen quality in a human population with idiopathic infertility. The gene encoding the circadian locomotor output cycles kaput protein (CLOCK) functions as an important positive enhancer of the circadian system. The observations reported in recent experimental animal studies suggested that CLOCK may play an important role in male reproduction. So far, such data for humans are not available. In this study, single-nucleotide polymorphisms (SNP) were used to examine the association between CLOCK and semen quality in human population with idiopathic infertility. Three-variant genotyping of CLOCK and semen analysis were performed in 478 males with idiopathic infertility by SNP genotyping assays and computer-assisted semen analysis. The results showed that the subjects carrying a C allele at rs3749474 (CC and TC) presented significantly lower semen volume compared with the TT genotype. For subjects carrying the CC genotype, sperm number per ejaculate and sperm motility were significantly lower compared with TT genotype carriers. The rs1801260 TC genotype carriers also had significantly lower sperm motility compared with the TT genotype. For the rs3817444 genotypes, the CA and AA genotype carriers presented significantly lower semen volume compared with the CC genotype. The findings suggested, as far as is known for the first time, an association between CLOCK genetic variants and altered semen quality in a human population with idiopathic infertility.

Surveillance of long-term environmental elements and PM2.5 health risk assessment in Yangtze River Delta, China, from 2016 to 2020.

PM2.5 metal pollution significantly harms human health. The air quality in Wuxi is poor, especially in winter, and long-term monitoring of PM2.5 elements comprising has not been performed previously. In the present study, 420 PM2.5 samples were collected from January 2016 to December 2020. Eleven elements, including Al, Mn, Ni, Cr, As, Cd, Sb, Hg, Pb, Se, and Tl, were analyzed by inductively coupled plasma mass spectrometry. The mean PM2.5 level was 56.1 ± 31.0 µg/m3, with a tendency of yearly decreasing and a significant seasonal distribution variation. The concentration of 11 elements in the PM2.5 samples was 0.38 ± 0.33 µg/m3. Al was the highest element with a range of 37.5-2148 ng/m3. Meanwhile, the spatial distribution differences were compared by literatures review. Based on the Crystal Ball model, health risks were assessed dynamically using Monte Carlo uncertainty analysis. After 10,000 simulations, the mean value of the hazard index for nine elements was 0.743, and Mn contributed the most to the hazard index among elements, with a correlation of 0.3464. The average carcinogenic risk was 1.01 × 10-5, which indicated that the non-carcinogenic and carcinogenic risks were within the acceptable range. However, considerable attention should be paid to the potential health risks associated with long-term Al, Mn, and As exposure. This study provides detailed data on local atmospheric pollution characteristics, helps identify potential risk elements, and contributes to the development of effective regional air quality management.

siRNA-mediated Eppin testicular silencing causes changes in sperm motility and calcium currents in mice.

Epididymal protease inhibitor (EPPIN) is differentially expressed in the reproductive tissues (such as testicles, outlet tubes, epididymis, vas deferens, and seminal vesicles). Its critical role in sperm function and male reproduction has shed light on EPPIN as a candidate target for male contraceptive vaccines. In this study, we endeavored to further reveal the mechanism through which EPPIN exerts its function. We created a mouse model of reduced Eppin expression by microinjecting small interfering RNA targeting Eppin expression into seminiferous tubules of mice. This mouse model was then used to explore the effects of low Eppin expression on sperm function, which was assessed by Computer Assisted Semen Analysis and patch clamp recording of T-type Ca2+ current in spermatogenic cells. We found that the sperm motility significantly declined when Eppin was downregulated. Further investigation demonstrated that Eppin downregulation significantly affected T-type Ca2+ currents and messenger RNA expression of three subtypes of T-type Ca2+ channels in spermatogenic cells. These findings indicate that low Eppin gene expression induces decreased T-type Ca2+ currents and mRNA expression, which in turn results in the reduced sperm motility.

The effect of anti-eppin antibodies on ionophore A23187-induced calcium influx and acrosome reaction of human spermatozoa.

BACKGROUND: Before a spermatozoon can fertilize an oocyte it must undergo a cascade of biochemical and physiological changes that facilitate its binding and penetration into the oocyte. Epididymal protease inhibitor (eppin) has been found to play a critical role in male fertility through an immunological approach. METHODS: In this study, we used an anti-eppin antibody to clarify the effect of eppin on human sperm functions during fertilization. Immunofluorescence studies were performed on ejaculated human spermatozoa in uncapacitated, capacitated and ionophore-treated states. Human spermatozoa were incubated in the presence or absence of anti-eppin antibody under capacitating conditions and with A23187. The effects of the antibody were evaluated on sperm motility, protein phosphotyrosine content and free intracellular calcium. RESULTS: Immunofluorescence results demonstrated that eppin is located on the acrosome and tail. After the acrosome reaction eppin is found on the equatorial segment and tail. We found that blocking eppin with antibodies significantly inhibited the human sperm acrosome reaction induced by A23187 in a dose-dependent manner. Finally, fluo-3 analysis demonstrated that the A23187-induced elevation of sperm intracellular calcium concentration was markedly reduced after incubation with anti-eppin antibody. However, the tyrosine phosphorylation of sperm proteins did not change. CONCLUSION: These results demonstrate that eppin can modulate intracellular calcium concentrations and subsequently affect the calcium ionophore A23187-induced acrosome reaction.

Variants of the EPPIN gene affect the risk of idiopathic male infertility in the Han-Chinese population.

BACKGROUND: It has been identified that human epididymal protease inhibitor (EPPIN) plays a critical role in sperm function and male fertility. The aim of this study was to determine whether variants of the EPPIN gene are risk factors for idiopathic male infertility. METHODS: All subjects, including 473 idiopathic infertile men and 198 fertile controls, underwent complete historical and physical examinations. Each subject donated 5 ml of peripheral blood for genomic DNA extraction and serum testosterone evaluation and an ejaculate for semen analysis. The semen analysis was performed by computer-assisted semen analysis system. The serum testosterone level was evaluated by radioimmunoassay. Four tagging single-nucleotide polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: We have demonstrated a significant decreased risk of idiopathic infertility with abnormal semen parameters in association with the variant rs2231829, and an increased risk of idiopathic infertility with abnormal semen parameters in association with the variant rs11594. However, among men with normal semen parameters, there were no differences in risk for these genotypes. Furthermore, no significant differences were found for the other variants, rs6124715 and rs2227290, on the risk of male infertility with normal or abnormal semen parameters. Similar serum testosterone levels among different EPPIN genotypes were observed for each group. CONCLUSIONS: These results suggest that different variants in the EPPIN gene may have different relationships with idiopathic male infertility and men carrying these variants have a decreased or increased risk of abnormal semen parameters associated with male infertility.

Variants in the Eppin gene show association with semen quality in Han-Chinese population.

The human epididymal protease inhibitor (Eppin) gene is specifically expressed in testis and epididymis and is important in male reproduction. However, to date, there is no report on variants of this gene, particularly in relation to male fertility. To investigate the association between Eppin genetic variants and semen quality, variant genotyping and semen analysis was performed in 473 males with definite idiopathic infertility by polymerase chain reaction-restriction fragment length polymorphism and computer-assisted semen analysis. It was found that rs6124715 GG/CG genotypes were associated with a significantly higher curvilinear velocity (VCL) (P=0.029 and 0.021 respectively) and average path velocity (VAP) (P=0.043 and 0.016 respectively) compared with the CC genotype. The straight line velocity (VSL) between rs6124715 CG and CC genotype was also significantly different (P=0.019). Regarding variant rs11594, subjects with C allele (CC or AC) had significantly lower VCL (P=0.011 and 0.046 respectively), VSL (P=0.025 and 0.041 respectively) and VAP (P=0.026 and 0.030 respectively) in comparison with AA homozygote. The sperm number per ejaculate was also significantly different between rs2231829 TT and CC genotype (P=0.042). These findings indicate, for the first time, that the genetic variants in the Eppin gene may be associated with semen quality.

Determination of 10 Haloacetamides in drinking water by gas chromatography with automated solid phase extraction.

Drinking water disinfection may result in the formation of different classes of toxic disinfection by-products (DBPs). Haloacetamides (HAcAms) are an emerging class of nitrogenous DBPs (N-DBPs), which are generally more prevalent at lower concentrations in disinfected water than carbonaceous DBPs. Herein a fast, convenient, and effective method of analyzing 10 HAcAms in drinking water samples was demonstrated. This method was developed using gas chromatography /electron capture detection (GC/ECD) supplemented with automated solid phase extraction (auto-SPE). The variables for automated SPE procedures were further optimized, including the selection of SPE sorbents, types and volumes of extraction solvents, SPE washing solvents and wash times. Under optimized conditions, the instrumental linearity range was 0.5-150 µg L-1 with correlation coefficients>0.9975. The limits of detection and quantification of this method were 0.002-0.003 µg L-1 and 0.005-0.010 µg L-1, respectively. The recovery values ranged from 72.4% to 108.5%, and the relative standard deviations ranged from 3.3% to 9.1%. Therefore, the auto-SPE-GC-ECD method showed acceptable linearity and repeatability and was subsequently validated and applied to analyze 10 HAcAms in drinking water.