Oxidative stress and DNA damage status in couples undergoing in vitro fertilization treatment.

This study examined the status of oxidative stress in 599 couples undertaking in vitro fertilization (IVF) treatment and its association with reproductive hormones, smoking, and outcomes. Oxidative stress biomarkers such as malondialdehyde, 8-hydroxy-2-deoxyguanosine, hydrogen peroxide (H2O2), catalase (CAT), and total antioxidant capacity (TAC) were determined in follicular fluid and seminal plasma. Tail moment (TM) was used to evaluate DNA damage in the sperm and granulosa cells. Reproductive hormones in serum and cotinine (COT) in urine, follicular fluid, and seminal plasma samples were determined. Separate multivariate linear regression was used to assess associations between levels of each oxidative stress biomarker and each hormone and smoking parameter (modeled as natural log-transformed). The findings indicate that some oxidative stress and DNA damage biomarkers played a role in disrupting certain reproductive hormones in women and their male partners either by overproducing reactive oxygen species or reducing antioxidant defense capacity. Although women were nonsmokers, COT levels > 50 and 10 µg/L in urine and follicular were observed in 5.7 and 1.7%, respectively. Levels of follicular fluid COT were positively associated with H2O2 and TM. We used log-binomial multivariate regression to estimate relative risks for the association between oxidative stress/DNA damage and IVF binary outcomes (fertilization rate > 50%, biochemical pregnancy, clinical pregnancy, and live birth). An increase in the CAT levels of follicular fluid was associated with a 48 and 41% decrease in the risk of poor fertilization rate (≤50%) and unsuccessful live birth, respectively. After the models were adjusted for hormonal factors, the associations remained the same, except that the elevated TAC in follicular fluid became significantly associated with a decrease of 42% in the risk of poor fertilization rate (≤50%). The higher antioxidant activity (CAT and TAC) in follicular fluid might positively impact specific IVF outcomes. LAY SUMMARY: Oxidative stress occurs when antioxidant molecules are insufficient in the body to destroy free radicals that can damage the cells, proteins and DNA, causing different health conditions, including infertility. The role of oxidative stress in female infertility has not received as much attention as male infertility, and research is still limited. This study explored whether the overproduction of free radicals can impact the success of in vitro fertilization (IVF) treatment using several biological markers such as hydrogen peroxide, catalase, and total antioxidant capacity. Our findings revealed that the high antioxidant levels in the fluid surrounding the egg were linked with a high fertilization rate. Additionally, oxidative stress status in couples was associated negatively with several reproductive hormones and smoking status. Biomarkers of oxidative stress and DNA damage might have potential applications in evaluating IVF patients' clinical characteristics such as causes of infertility, hormonal profile, fertilization rate, implantation and live birth.

Effects of early and recent mercury and lead exposure on the neurodevelopment of children with elevated mercury and/or developmental delays during lactation: A follow-up study.

This follow-up study of 82 children investigated the potential impact of early and recent exposure to mercury and lead on their neurodevelopmental performance at 5-8 years of age (2017-2018). Early exposure of these children to mercury, methylmercury, and lead was assessed during lactation at 3-12 months old, as well as their mother's exposure using measurements from a cross-sectional study (2011-2013). Only infants who failed to pass the neurodevelopment screening tools and/or had elevated mercury were included in this study. Urine and hair were sampled during the follow-up study to assess the children's recent exposure to mercury, methylmercury, and lead. Their cognitive performance and visual-motor integration were also measured using the Test of Non-Verbal Intelligence (TONI) and the Beery-Visual-Motor Integration (Beery VMI), respectively. The association between alterations in urinary porphyrins excretion and exposure to metals was analyzed and their influence on the children's neurodevelopment was explored. Linear regression models revealed a significant negative association between the infants' mercury exposure during lactation and the TONI Quotient (ß = -0.298, 95%CI = -4.677, -0.414) and Beery VMI Age Equivalent scores at age 5-8 (ß = -0.437, 95%CI = -6.383, -1.844). The mothers' blood methylmercury was inversely and significantly associated with their children's TONI Quotient (ß = -0.231, 95%CI = -8.184, -0.331). In contrast, the children's Beery VMI Age Equivalent scores were positively and significantly associated with the hair methylmercury of the mothers (ß = 0.214, 95%CI = 0.088, 3.899) and their infants (ß = 0.256, 95%CI = 0.396, 4.488). These relationships suggest the presence of negative confounding that we did not take into account. Unlike mercury, there was some evidence that lead in breast milk had an inverse relationship with the children's visual-motor coordination skills. Our study did not show a clear association between children's recent exposure to metals and neurodevelopment. However, a significant inverse association was observed between the TONI Quotient and the interaction of hair methylmercury × ∑porphyrins (ß = -0.224, 95%CI = -0.86, -0.049), implying that porphyrins are a sensitive measure of low body-mercury burden. Although lead induced higher ∑porphyrins excretion in urine (ß = 0.347, 95%CI = 0.107, 0.525), their interaction did not influence children's neurodevelopmental scores. The interactions between metals and porphyrins might provide insights into their potential contributory role in the pathogenesis associated with neurological disorders or other diseases. Despite the small sample size of the present study, its findings about the association between toxic metal exposure and the high risk of poor neurodevelopmental performance are worrying, particularly at an early age, and additional research is needed using larger sample sizes.

Cytotoxic and genotoxic effects of e-liquids and their potential associations with nicotine, menthol and phthalate esters.

In this study, we determined DNA damage and chromosome breakage (indicators of genotoxicity) and cell viability (an indicator of cytotoxicity) in human lymphoblastoid TK6 and Chinese hamster ovary (CHO) cells treated with 33 e-liquids using in vitro single cell gel (comet), micronucleus (MN), and trypan blue assays, respectively. We also measured the contents of nicotine, five phthalate esters, and DL-menthol in the e-liquids to examine their effects on DNA damage, chromosome breakage, and cell viability. Our chemical analyses showed that: (1) six e-liquids had nicotine ≥2-fold higher than the manufacture's label claim (2-3.5 mg); (2) both dimethyl- and dibutyl-phthalate levels were >0.1 µg/g, i.e., their threshold limits as additives in cosmetics; and (3) the DL-menthol contents ranged from 0.0003 to 85757.2 µg/g, with those of two e-liquids being >1 mg/g, the threshold limit for trigging sensory irritation. Though all the e-liquids induced DNA damage in TK6 cells, 20 resulted in cell viabilities ≤75%, indicating cytotoxicity, yet the inverse relationship between cell viability and DNA damage (r = -0.628, p = 0.003) might reflect their role as pro-apoptotic and DNA damage inducers. Fifteen e-liquids induced MN% in TK6 cells ≥3-fold that of untreated cells. Some of the increase in %MN might be false due to high cytotoxicity, yet six brands showed acceptable cell viabilities (59-71%), indicating chromosome damage. DNA damage and %MN increased when the TK6 cells were exposed to metabolic activation. The CHO cells were less sensitive to the genotoxic effects of the e-liquids than the TK6 cells. DL-menthol was found to be associated with decreased cell viability and increased DNA damage, even at low levels. We cannot dismiss the presence of other ingredients in e-liquids with cytotoxic/genotoxic properties since out of the 63 different flavors, 47 induced DNA damage (≥3-folds), and 26 reduced cell viability (≤75%) in TK6 cells.

Robust expression of tumor suppressor miRNA's let-7 and miR-195 detected in plasma of Saudi female breast cancer patients.

BACKGROUND: Female breast cancer is frequently diagnosed at a later stage and the leading cause of cancer deaths world-wide. Levels of cell-free circulating microRNAs (miRNAs) can potentially be used as biomarkers to measure disease progression in breast cancer patients in a non-invasive way and are therefore of high clinical value. METHODS: Using quantitative RT-PCR, circulating miRNAs were measured in blood samples collected from disease-free individuals (n = 34), triple-negative breast tumours (TNBC) (n = 36) and luminal tumours (n = 57). In addition to intergroup comparisons, plasma miRNA expression levels of all groups were analyzed against RNASeq data from cancerous breast tissue via The Cancer Genome Atlas (TCGA). RESULTS: A differential set of 18 miRNAs were identified in the plasma of breast cancer patients and 10 miRNAs were uniquely identified based on ROC analysis. The most striking findings revealed elevated tumor suppressor let-7 miRNA in luminal breast cancer patients, irrespective of subtype, and elevated miR-195 in plasma of TNBC breast cancer patients. In contrast, hsa-miR-195 and let-7 miRNAs were absent from cancerous TCGA tissue and strongly expressed in surrounding non-tumor tissue indicating that cancerous cells may selectively export tumor suppressor hsa-miR-195 and let-7 miRNAs in order to maintain oncogenesis. CONCLUSIONS: While studies have indicated that the restoration of let-7 and miR-195 may be a potential therapy for cancer, these results suggested that tumor cells may selectively export hsa-miR-195 and let-7 miRNAs thereby neutralizing their potential therapeutic effect. However, in order to facilitate earlier detection of breast cancer, blood based screening of hsa-miR-195 and let-7 may be beneficial in a female patient cohort.

Alterations in biochemical markers due to mercury (Hg) exposure and its influence on infant's neurodevelopment.

This study examined the role of oxidative stress due to mercury (Hg) exposure on infant's neurodevelopmental performance. A total of 944 healthy Saudi mothers and their respective infants (aged 3-12 months) were recruited from 57 Primary Health Care Centers in Riyadh City. Total mercury (Hg) was measured in mothers and infants urine and hair samples, as well as mother's blood and breast milk. Methylmercury (MeHg) was determined in the mothers and infants' hair and mother's blood. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), malondialdehyde (MDA), and porphyrins were used to assess oxidative stress. The infant's neurodevelopment was evaluated using Denver Developmental Screening Test II (DDST-II) and Parents' Evaluation of Developmental Status. The median total Hg levels in mother's urine, infant's urine, mother's hair, infant's hair, and mother's blood and breast milk were 0.995µg/l, 0.716µg/l, 0.118µg/g dw, 0.101µg/g dw, 0.635µg/l, and 0.884µg/l respectively. The median MeHg levels in mother's hair, infant's hair, and mother's blood were 0.132µg/g dw, 0.091µg/g dw, and 2.341µg/l respectively. A significant interrelationship between mothers and infants Hg measures in various matrices was noted. This suggests that mother's exposure to different forms of Hg (total and/or MeHg) from various sources contributed significantly to the metal body burden of their respective infants. Even though Hg exposure was low, it induced high oxidative stress in mothers and infants. The influence of multiplicative interaction terms between Hg measures and oxidative stress biomarkers was tested using multiple regression analysis. Significant interactions between the urinary Hg levels in mothers and infants and oxidative stress biomarkers (8-OHdG and MDA) were noted. The MeHg levels in mother-infant hair revealed similar interaction patterns. The p-values for both were below 0.001. These observations suggest that the exposure of our infants to Hg via mothers either during pregnancy and/or neonatal life, promoted oxidative stress that might have played a role in infant neurodevelopmental delays that we reported previously. The results confirmed that the interaction between infant's MeHg in hair and 8-OHdG and MDA levels was significantly associated with a delay in DDST-II performance (ß=-0.188, p=0.028). This finding provides an insight into the potential consequences of Hg-induced oxidative stress to infant's cognitive neurodevelopment for the first time. This observation still needs future studies to be validated. Given the low MeHg levels in our population, these findings are of particular importance.

Mercury (Hg) exposure and its effects on Saudi breastfed infant's neurodevelopment.

This cross-sectional study analyzed mercury (Hg) levels in healthy Saudi mothers and their infants (age 3-12 months) and examined the influence of Hg on the infants' neurodevelopment using screening tools, such as the Denver Developmental Screening Test II (DDST-II) and Parents' Evaluation of Developmental Status (PEDS). A total of 944 mothers and their 944 infants were recruited from 57 Primary Health Care Centers (PHCCs) in Riyadh. The total Hg (THg) levels were measured in the mothers' and infants' urine (UTHg-M and UTHg-I) and hair (HTHg-M and HTHg-I) samples and in the breast milk and mothers' blood. Methylmercury (MeHg) levels were determined in hair samples from the mothers (MeHg-M) and infants (MeHg-I). Only 40.1% of the infants were breast-fed when enrolled, and 59.9% had stopped breastfeeding. Only 1.8% of the mothers and 0.3% of the infants had MeHg levels above the Environmental Proection Agency (EPA) reference dose (1 µg/g), with low medians of 0.132 and 0.091 µg/g dw, respectively, but the MeHg levels were significantly associated with infant DDST-II performance. The levels of corrected UTHg-M for creatinine (Cr), HTHg-M, HTHg-I, and HMeHg-M, however, displayed an association with infant PEDS performance. The medians and percentage of the tested population that exceeded the recommended limits for Hg in urine and hair set by the World Health Organization (5 µg/g Cr) and EPA (1 µg/g) were 0.695 µg/g Cr and 3% UTHg, 0.118 µg/g dw and 4.1% HTHg-M, 0.101 µg/g dw and 2.8% HTHg-I, and 0.132 µg/g dw and 1.8% HMeHg-M. Our study provides evidence of an association between some Hg measures and delays in infant neurodevelopment, despite their low levels and regardless of the infant's breastfeeding status. The results are of potential concern, because delayed psychomotor or mental performance in infants could be an indicator of later neurocognitive development in children, which may persist into adulthood, as shown in other studies. The absence of local standardization of the DDST-II and PEDS screening tools might raise some questions, although the DDST-II has been used in local institutions for a number of years. The development of effective standardized developmental screening tools is necessary to ensure that all children at risk of neurodevelopmental problems early in life are identified so that they can receive appropriate and timely intervention.

The extent of mercury (Hg) exposure among Saudi mothers and their respective infants.

A total of 1016 healthy Saudi mothers and their respective infants (aged 3-12 months) were recruited from 57 Primary Health Care Centers (PHCCs) in Riyadh, Saudi Arabia, to evaluate the extent of mercury (Hg) exposure and predict its sources in the healthy Saudi population. Total Hg levels were measured in maternal urine, breast milk, blood, and hair and in the infants' urine and hair. Only 1.9% of the mothers had urinary Hg (UHg)>10 µg/l, the limit for asymptomatic adults recommended by the World Health Organization, but the median (0.99 µg/l) was higher than in other countries. Also, 49.3% of the mothers had UHg>1 µg/l, the German reference value for adults. Median infant UHg was 0.729 µg/l, and 77 and 93 % of the infants had levels higher than 0.4 and 0.1 µg/l, the reference values of the Centers for Disease Control and Prevention and for Germany, respectively. The median Hg level in breast milk was 0.884 µg/l. Even though 43.2% of the milk samples were above the background level for Hg in human milk (1 µg/l), our results were lower than those reported from other countries. Median maternal total Hg in blood was 0.637 µg/l, and only 0.4 and 6.9% of samples were higher than the Hg reference levels of 5.8 µg/l of the Environmental Protection Agency (EPA) and of 2 µg/l for Germany, respectively. Total Hg levels in hair (HHg) varied widely among mothers and infants, but only 3.9% of the mothers and 2.8% of the infants had HHg>1 µg/g (the EPA reference level). Median HHg values were 0.117 µg/g dry weight in mothers and 0.1 µg/g dry weight in infants; both were lower than in other countries. The Hg levels in mothers and their respective infants were relatively low, but our results were consistent with other studies indicating that dental amalgam fillings and fish consumption were the main predictors of maternal Hg exposure. Among the several biomarkers of Hg exposure, Hg levels in maternal hair and urine were the strongest predictors of infant exposure. The lack of an association between Hg in breast milk and Hg in infant urine and hair suggested that the infants were exposed to Hg predominately during pregnancy rather than during breastfeeding. We expect that our data can serve as a baseline for further biomonitoring and follow-up studies, particularly of the long-term impact of Hg on childhood neurodevelopment.