Tusha Sharma

Heavy Metal Levels in Adolescent and Maternal Blood: Association with Risk of Hypospadias

Background. Hypospadias is a part of testicular digenesis syndrome (TDS) which includes infertility, cryptorchidism, and spermatogenesis. Heavy metals act as endocrine disrupting compounds. Heavy metals such as cadmium, chromium, arsenic, and lead have been associated with male infertility, cryptorchidism, spermatogenesis, cancer, reproductive disorder, and neurological disorder. However, it remains an important issue to corroborate or refute the hypothesis that the role of heavy metals in male reproductive tract disorders. Hence, the present study was designed to investigate the possible association of heavy metal and risk of hypospadias by estimating the blood heavy metal levels. Methods. In this case control study, 50 hypospadias boys diagnosed and confirmed by a pediatric urologist and 50 randomly selected age-matched (1–5 years) healthy control boys not suffering from any clinically detectible illness and their mothers have been included and heavy metal levels in the blood of these subjects have been estimated by Atomic Absorption Spectrophotometer (AAS). Result. Significantly high levels of cadmium and lead have been observed in hypospadias cases; however, all heavy metal levels were present in higher concentration. Conclusion. Higher blood levels of cadmium and lead may be associated with the increased risk of hypospadias.

In Vitro Propagation of Endangered Temperate Himalayan Medicinal Herb Picrorhiza kurroa Royle ex Benth using Leaf Explants and Nodal Segments

The present study describes the micropropagation of Picrorhiza kurroa, (commonly known as kutki) an endangered medicinal herb of the temperate Himalayas and a source of hepatoprotective picrosides. In vitro shoot multiplication was achieved through sprouting of axillary buds using nodal segments and leaf tissue. For shoot regeneration, the hormone combinations kinetin (2.0 mg l−1) and Kinetin + Indole-3-butyric acid (IBA) (2.0 mg l−1 + 0.50 mg l−1) with leaf explant was found superior. Interestingly, the basal MS medium gave 99.94 % response (direct proliferation) with nodal explant. The medium supplemented with IBA (1.0 mg −1) was found best for rooting of regenerated shoots. Nodal segments plated on the medium supplemented with TDZ + IBA (0.11 mg −1 + 0.50 mg −1) formed somatic embryos, however further regeneration could not be achieved. The in vitro raised plantlets were hardened and successfully established in the glass house conditions.

Assessment of Toxicogenomic Risk Factors in Etiology of PretermDelivery

Worldwide, preterm birth accounts for more than 60% of prenatal morbidity and mortality and is the leading cause of neonatal deaths. Globally, every year, around 13 million babies are born preterm, with rates being highest in the low and middle-income group. India is leading all the nations in PTD, as reported by WHO. PTD is considered as a major pregnancy complication. PTD continues to be a major clinical and public health problem and is regarded as a syndrome with multiple causes. Environmental stressors such as heavy metal, Organochlorine pesticides have a long history of widespread use all around the world. Because of high stability, extremely low biodegradability and long half life, these compounds are typically very persistent in the environment, and are known to accumulate in soil, food, blood, fatty tissue etc. Humans, being at the top of the food chain are most vulnerable to health effects as the level of the toxic chemicals is several folds higher through the process of bio-magnifications. The problem gets compounded in women because of increased adipose tissue in them and high liposolubility of these OCPs. Pesticide toxicity is of utmost concern during pregnancy as studies have revealed that mother and fetus are more vulnerable to their toxic effects. OCPs are metabolized by xenobiotic metabolizing enzymes such as CYP P450 and GSTs. Polymorphism in xenobiotic metabolizing genes may cause improper metabolism of xenobiotics which may cause high free radical generation, increased oxidative stress, improper cytokines release and also increased inflammation. OCPs levels are also correlated significantly with increased expression of COX-2 gene. The mechanisms underlying the etiology and onset of preterm birth is not clearly understood. Still more than 40-50% cases of preterm delivery are ‘idiopathic’. So, it is very essential to assess the etiological factors and the mechanisms by which they result in preterm birth in order to make further research in targeted interventions to prevent preterm labor.

Organochlorine pesticide exposure in mothers and neural tube defects in offsprings

We explored the association between maternal exposure to organochlorine pesticides and neural tube defects (NTDs) in the offspring. Blood was collected from 35 mothers and their offsprings with NTDs (case group) and from 35 mothers-neonate dyads without congenital anomalies (control group). The median blood levels of DDE, t-HCH and endosulphan in mothers in the case group and of dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE), total hexachlorocyclohexane (t-HCH) and endosulfan in the neonates with NTDs were significantly higher. Neonates with NTDs had 3.6 times more chances of having blood levels of endosulfan above the median level of the control group. Mothers delivering offsprings with NTDs had 11.3 times greater chances of having DDE levels above the median concentration in the control group. We recommend a restrained use of organochlorine pesticides like DDT, DDE, and endosulfan, while monitoring the expectant mothers closely for birth defects like NTDs.

Role of Organochlorine Pesticides in Children with Idiopathic Seizures

Background. Organochlorine pesticides (OCP) are persistent organic pollutants that have been implicated in causing several deleterious effects in humans. These are known neurotoxins in high doses, but the role of environmentally acquired OCPs in the body to induce seizures in children has not been investigated yet. Objectives. To assess the serum levels of OCPs in children aged 2–12 with idiopathic seizure and to find out any association between the two are our objectives. Methods. It was a cross-sectional pilot study. Twenty developmentally normal children aged 2–12, presenting with idiopathic generalized seizures, were recruited. Twenty age-matched controls without any history of seizures were also taken. Their serum levels of α, β, and γ hexachlorocyclohexane (HCH); and aldrin; dieldrin; p,p-dichlorodiphenyltrichloroethane (DDT), o,p-DDT, and p,p dichlorodiphenyldichloroethylene (DDE); and α and β endosulfan were analysed using gas chromatography (GC). Mann-Whitney U test was used to compare OCP levels between the groups. Spearman correlation was used to find the correlation between individual pesticide levels with age and seizure duration. Results. Levels of β, γ, and total HCH were significantly higher among cases as compared to the control group (P ≤ 0.05). Conclusion. There exists a possible association between idiopathic seizures and high serum levels of OCPs, especially HCH.

Delineating Potential Transcriptomic Association with Organochlorine Pesticides in the Etiology of Epithelial Ovarian Cancer

Background: Recent studies have shown that there is an increased risk of Epithelial Ovarian Cancer (EOC) with Organochlorine Pesticides (OCPs). However, the alteration in the gene expression profile has not been explored so far. The goal of the present study is to understand the probable molecular mechanism of OCPs toxicity towards discovery of dysregulation of signaling pathway associated with differential gene expression and candidate transcriptomic set of markers in the pathophysiology of EOC in OCPs exposed population. Methods: The OCP levels were estimated by gas chromatography and whole genome differential expression study was carried out using expression microarray and candidate genes were validated using Real time RT-PCR. Results: Significant level of OCP residues such as β-hexachlorocyclohexane (β-HCH), Heptachlor, Heptachlor epoxide B (HTEB), dichlorodiphenyldichloroethylene (p’p’-DDE) and endosulfan-I was found between healthy and EOC patients. The transcriptome profile of several genes revealed regulation of various important cellular processes such as metabolism, inflammation, cytoskeleton dysregulation of TGF and WNT pathway in EOC cases with high OCPs. Conclusion: This study provides the first evidence showing that differentially expressed genes and dysregulation of signaling pathways might be associated with significant level of OCPs exposure in ovary tissue of epithelial ovarian cancer patients. Moreover, significant correlation of these genes with OCPs revealed that OCPs exposure played vital role in dysregulation of related pathways in the etiology of EOC

Modulation of serum BDNF levels in postherpetic neuralgia following pulsed radiofrequency of intercostal nerve and pregabalin.

AIM To study the modulation of serum BDNF levels following integrated multimodal intervention in postherpetic neuralgia (PHN). MATERIALS & METHODS A randomized, double-blind controlled study was undertaken among patients of thoracic PHN where 30 patients received pregabalin with pulsed radiofrequency and 30 controls received pregabalin with sham treatment. RESULTS Pain intensity (visual analog scale) was reduced earlier in intervention group (15.3 ± 5.7 at the fourth week) compared with control group (16.3 ± 6.6 at the eighth week). Serum BDNF level increased with time in both the groups with overall increase more pronounced in intervention group. CONCLUSION Integrated multimodal therapy using minimally invasive pulsed radiofrequency and pregabalin in PHN was effective in early pain reduction with elevated serum BDNF levels.

Association of organochlorine pesticides with the mRNA expression of tumour necrosis factor-alpha (TNF-α) & cyclooxygenase-2 (COX-2) genes in idiopathic preterm birth.

Background & objectives: Preterm birth (PTB) is an important cause of prenatal death, neonatal morbidity and mortality and adult illness. Increased inflammation occurs in normal parturition, and inflammatory cytokines and oxidative stress are found to be higher in PTB cases. The present study was planned to investigate the association of organochlorine pesticides (OCPs) with mRNA expression of inflammatory pathway genes such as tumour necrosis factor-alpha (TNF-α) and cyclooxygenase-2 (COX-2) in preterm delivery (PTD) cases. Methods: Maternal blood samples of PTD (n=30) cases and equal number of term delivery (n=30) were collected at the time of labour. Women occupationally exposed to OCPs and other high risk factors such as anaemia, hypertension, bacterial vaginosis, renal and heart disease, diabetes, etc. were excluded. The OCP levels were estimated by gas chromatography, and mRNA expressions of TNF-α and COX-2 genes were analysed using real-time PCR (qPCR). Results: Significantly higher levels of β-HCH (beta-hexachlorocyclohexane, 95% CI=2.08-4.633, P=0.001), p’p’-DDE (para, para-dichlorodiphenyldichloroethylene, 95% CI=0.546-2.551, P=0.003), and o’p’-DDD (ortho, para-dichlorodiphenyldichloroethane, 95% CI=0.004-0.690, P=0.047) were observed in maternal blood of PTB cases as compared to term delivery. The mRNA expressions of COX-2 and TNF-α genes were 3.13 and 2.31 folds higher in PTB cases in comparison to term delivery. Linear positive correlations were observed between period of gestation (POG) and ΔCt of COX-2 and TNF-α genes. Interpretation & conclusions: Environmental factors such as OCPs may be associated with inflammatory events showing gene-environment interaction in PTB cases. Evaluating the molecular control of inflammation along with gene environment interaction may be used as a model to explore the aetiology of idiopathic PTB cases and may be considered for the prognosis of adverse reproductive outcomes.

CYP1A1 expression and organochlorine pesticides level in the etiology of bladder cancer in North Indian population

Urinary bladder cancer (UBC) is one of the most common malignancies worldwide. The etiology of UBC is multifactorial and includes both exogenous and endogenous factors. Exogenous risk factors include exposure to heavy metals, aromatic amines, and environmental pollutants including pesticides such as organochlorine pesticides (OCPs). Environmental factors alone are incapable of directly producing DNA damage and may require activation by phase I metabolizing enzymes like cytochrome P450 in order to become active carcinogen. The present study is designed to study CYP1A1 gene expression, OCP level in cases of UBC, as well as to explore the plausible role of gene–environment interaction in the etiology of UBC among North Indian population. A total of 60 cases with equal number of controls were enrolled under this study, the OCP levels were estimated using gas chromatography, CYP1A1 mRNA expression was quantified by real-time quantitative polymerase chain reaction, and fold change was calculated using the ΔΔCt method. In the present study, the levels of OCP were found to be significantly higher with the upregulation of CYP1A1 mRNA expression among UBC cases as compared to controls. While putting multiple linear regression, it has been observed that there is a significant interaction between the levels of OCPs and ΔCt value of CYP1A1 gene taken into account hematuria episodes as dependent variable. The study concludes that when there is predisposition of OCPs and upregulation of CYP1A1 gene, then the result will be an increment in hematuria episodes which indicates that gene–environment interaction plays a significant role in the causation of UBC among North Indian population.

Assessment of urinary cotinine levels in women with gynecological complaints at a tertiary care hospital: A pilot study

Gynecological effects due to smokeless tobacco exposure are not well studied. This cross-sectional study was undertaken with the objective to evaluate the urinary cotinine levels in women of reproductive age with gynecological complaints. The study was conducted in 2015 at the outpatient clinic of the Department of Obstetrics and Gynecology, University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi. A total of 192 consecutive women presenting with gynecological complaints (pelvic inflammatory disease (PID), infertility, and menstrual abnormality) were recruited. Their demographic details and tobacco exposure were recorded. All of them denied exposure to any form of tobacco. Urinary cotinine level of each participant was measured. The mean urinary cotinine level was 23.60 ± 12.00 ng/ml. PID was the most common gynecological complaint. Women with PID had significantly higher urinary cotinine levels compared to those with menstrual complaints and infertility: 24.9548 (±12.259) ng/ml versus 20.2042 (±10.9248) ng/ml. This study highlights the importance of addressing the issue of secondhand smoke exposure and reproductive morbidities in women.