Neeraj Sinha

Therapeutic targets of triple-negative breast cancer: a review.

Breast cancer (BC) is the second most common cause of cancer deaths. Triple‐negative breast cancer (TNBC) does not show immunohistochemical expression of oestrogen receptors, progesterone receptors or HER2. At present, no suitable treatment option is available for patients with TNBC. This dearth of effective conventional therapies for the treatment of advanced stage breast cancer has provoked the development of novel strategies for the management of patients with TNBC. This review presents recent information associated with different therapeutic options for the treatment of TNBC focusing on promising targets such as the Notch signalling, Wnt/β‐catenin and Hedgehog pathways, in addition to EGFR, PARP1, mTOR, TGF‐β and angiogenesis inhibitors.

Chronic toxicity of styrene maleic anhydride, a male contraceptive, in rhesus monkeys (Macaca mulatta)

A newly developed male contraceptive, styrene maleic anhydride (SMA), was injected in the vas deferens of male rhesus monkeys for safety evaluation at the dose of 100 mg (contraceptive dose, CD), 250 mg (CD x 2.5) and 500 mg (CD x 5.0), and the monkeys were kept under observation for one year. The observed behavioural, haematological, biochemical and histopathological parameters in treated monkeys were comparable to controls. The results suggest the polymer SMA to be safe up to 5 times CD in monkeys.

Role of Apoptosis in Mediating Salicylic Acid-Induced Teratogenesis In Vitro

Salicylic acid (SAL) is among the most commonly used drugs worldwide and is known to cause congenital malformations and fetal death in animals. In this study, the effect of SAL on major organogenesis period and the role of apoptosis in mediating congenital malformations have been carried out. In the present study, post-implantation rat embryos of day 11 were cultured for 24 h with various concentrations of SAL, i.e. 10, 100, and 1000 μg/ml cultures, respectively. The growth and developmental of each embryo was evaluated and compared with control ones for the presence of any malformations. The SAL decreased all growth and developmental parameters in a concentration-dependent manner, when compared with control. However, exposure to SAL at 10 μg/ml culture did not show any significant effect on embryonic growth and development. Parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were carried out followed by quantitation by 3′-OH labeling of cultured rat embryos to evaluate the role of apoptosis in bringing about SAL-induced teratogenesis. All results were found to be dose-dependent and an increase in apoptosis in embryonic tissues may be related to the increased risk of congenital malformations. The data suggested that apoptosis might be involved in mediating teratogenesis of SAL in vitro.

Involvement of apoptosis in mediating mitomycin C-induced teratogenesis in vitro

Mitomycin C (MMC) is among the most commonly used drugs worldwide and is known to cause congenital malformations and fetal death in animals. In this study, the effect of MMC on major organogenesis period and the role of apoptosis in mediating congenital malformations have been carried out. In the present study, post-implantation rat embryos of day 11 were cultured for 24u2009h with various concentrations of MMC, i.e. 1, 10, and 100 µg/ml cultures. The growth and developmental of each embryo was evaluated and compared with control ones for the presence of any malformations. The MMC decreased all growth and developmental parameters in a concentration-dependent manner, when compared with control. However, exposure to MMC at 1 µg/ml culture did not show any significant effect on embryonic growth and development. Parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were carried out followed by quantitation by 3′-OH labeling of cultured rat embryos to evaluate the role of apoptosis in bringing about MMC-induced teratogenesis. All results were found to be dose-dependent and an increase in apoptosis in embryonic tissues may be related to the increased risk of congenital malformations. The data suggested that apoptosis might be involved in mediating teratogenesis of MMC in vitro.

Detecting Role of Apoptosis in Mediating Cyclophosphamide Induced Teratogenesis In Vitro

Programmed cell death (apoptosis) refers to a specific type of cell death under stringent genetic control. Even a slight alteration in this process leads to malformations characterized by birth defects. Based on the above hypothesis we deduced that apoptosis plays an important role in mediating the teratogenicity of cyclophosphamide in vitro. The present study was undertaken to see whether this phenomenon holds true or not. In this study, 11-day-old rat embryos were cultured for 24 hours with various concentrations of CP (i.e. 0, 5, 10 and 100 μg/ml culture). After culturing for 24 hours, embryos exposed to 10 and 100 μg/mL culture of CP were found having both malformations and growth retardation. Exposure to CP at 5 μg/mL culture did not show significant effect on embryonic development. Parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were also carried out followed by quantitation by 3′-OH labeling of cultured embryos to evaluate CP-induced apoptosis. All the results were found to be dose-dependant, and the data suggested that apoptosis is involved in mediating the teratogenicity of CP in vitro.

Studying significance of apoptosis in mediating tolbutamide-induced teratogenesis in vitro.

The incidence of type 2 diabetes mellitus (non‐insulin‐dependent diabetes mellitus) is growing worldwide and poses a serious public health problem in a current paradigm of changing life style and food habits. Tolbutamide (sulfonylurea) is among the commonly used anti‐diabetic drugs worldwide for treating type 2 diabetes and is known to cause congenital malformations in animals. In this study, the effect of tolbutamide on major organogenesis period and the possible involvement of apoptosis in mediating congenital malformations have been carried out. In the present study design, post‐implantation rat embryos of day 11 were cultured for 24u2003h with various concentrations of tolbutamide, i.e., 10, 100, and 1000u2003μg/mL cultures, respectively. The growth and developmental of each embryo was evaluated and compared with control ones for the presence of any malformations. The tolbutamide decreased all growth and developmental parameters in a concentration‐dependent manner, when compared with control. However, exposure to tolbutamide at 10u2003μg/mL culture did not show any significant effect on embryonic growth and development in vitro. In parallel to this, flow cytometric analysis (cell cycle and annexin V binding) and DNA fragmentation assay were carried out followed by quantitation by 3′‐OH labeling of cultured rat embryos to examine the role of apoptosis in bringing about tolbutamide‐induced teratogenesis. All results were found to be dose dependent and an increase in apoptosis in embryonic tissues may be related to the increased risk of congenital malformations. The outcome of the research suggested that apoptosis might be involved in mediating teratogenesis of tolbutamide in vitro. Further research is warranted to fully understand this mechanism.

Metabolic fingerprinting in breast cancer stages through 1H NMR spectroscopy-based metabolomic analysis of plasma

HIGHLIGHTSMetabolic fingerprints in the blood plasma of early and late stage breast cancer were analyzed by 1H‐NMR spectroscopy based metabolomics.Sixteen metabolites were found be differentially modulated in early and late stage breast cancer as compared with healthy subjects.Hydroxybutyrate, lysine, glutamate, glucose, lactate, and NAG levels were significantly altered with good biomarker potential in breast cancer.Glutamate, Lactate and NAG showed a good classifier for advanced stage breast cancer progression. ABSTRACT Breast cancer (BC) is one of the most common malignancies among women worldwide, which is indeed associated with metabolic reprogramming. However, BC is a very complex and heterogeneous disease, which can relate with the changes in metabolic profiles during BC progression. Hence, investigating the metabolic alterations during BC stage progression may reveal the deregulated pathways and useful metabolic signatures of BC. To demonstrate the metabolic insights, we opted 1H NMR spectroscopy based metabolomics of blood plasma of early and late stage BC (N = 72) with age and gender matched healthy subjects (N = 50). Further, the metabolic profiles were analyzed to delineate the potential signatures of BC by performing multivariate and nonparametric statistical analysis in early and late stages of BC in comparison with healthy subjects. Sixteen metabolites levels were differentially changed (p < 0.05) in the early and late stages of BC from healthy subjects. Among them, the levels of hydroxybutyrate, lysine, glutamate, glucose, N‐acetyl glycoprotein, Lactate were highly distinguished in BC stages and showed a good biomarker potential using receiver‐operating curves based diagnostic models. Furthermore, the significant modulation and good diagnostic performances of glutamate, N‐acetyl glycoprotein and Lactate in LBC as compared to EBC give their significance in the BC progression. In general, our observations demonstrate that these panels of metabolites may act as vital component of the metabolism of early to late stage BC progression. Our results also open new avenue towards early and late stage BC diagnosis and intervention implying metabolomics approaches.

Role of apoptosis in mediating diclofenac-induced teratogenesis: An in vitro approach.

Diclofenac (DCF) is among the most commonly used nonsteroidal anti-inflammatory drugs worldwide for the treatment of various conditions in postpubertal women. However, very limited information is available regarding its safety during pregnancy and teratogenecity. The present study was designed to elucidate the effects of DCF on the developing rat embryos during the major organogenesis period and investigate the critical role of apoptosis in bringing about these congenital anomalies. Embryos were exposed in vitro to various concentrations of DCF, that is, 0, 3.75, 7.5 and 15 µg/ml for 24 h, respectively, and examined for the growth and differentiation at the end of the culture period for the presence of any specific malformations. Growth and developmental parameters such as weight of embryos, crown–rump length and number of somites were found to be lower in the embryos exposed to high concentrations of DCF (7.5 and 15.0 μg/ml) when compared with the untreated control. However, no significant difference in growth parameters was found between embryos exposed to 3.75 µg/ml and the control group. In parallel to this, flow cytometric analysis and DNA quantitation of cultured rat embryos were performed to verify the involvement of apoptosis in mediating DCF-induced teratogenesis. A concentration-dependent increase in apoptosis in embryos suggests a possible engagement of apoptosis in the role of DCF as a teratogenic agent. A detailed analysis of the actual effect of DCF on cellular apoptotic machinery necessitates further evaluation.

A novel approach for testing the teratogenic potential of chemicals on the platform of metabolomics: studies employing HR-MAS nuclear magnetic resonance spectroscopy

NMR based metabolomics offers a complementary approach that gives information on whole-organism functional integrity over time after drug exposure. Hence the objective was to develop a quick, reliable method for testing the teratogenic potential of a new chemical entity (NCE) on the platform of metabonomics, as an alternative to conventional procedures. Time mated Charles Foster rats (nxa0= 9) were injected with cyclophosphamide (0, 5, 15 and 30 mg kg−1) at different dose levels on day 11 of the pregnancy, through an i.p. route. On day 12 of the pregnancy, embryos were procured from six rats out of the 9 pregnant rats from each group, using a per abdominal approach. These embryos were then undertaken for morphological studies and NMR experiments using a high resolution-magic angle spin (HR-MAS) probe. The remaining three rats were followed until term to procure full term pups, which were used for conventional observations and studies. Multivariate unsupervised principal component analysis of the 1H NMR spectra from the rat embryos revealed a dose dependent cluster separation between the controls and treated specimens, which was further confirmed by supervised partial least-squares discriminant analysis with an R2 of 0.77 and Q2 of 0.72. Those embryos which were found to have malformations/anomalies, significantly presented a few upregulated (aspartate) and a few downregulated (creatine, choline and glycine) metabolite levels. This work revealed that observed teratogenic ailments and resulting metabolic profiles have a definite correlation, demonstrating the suitability of this procedure for testing teratogenicity, which may give a new dimension to the prioritization of lists having a large number of chemicals to be tested for their teratogenicity. Malformations observed among the full term pups also validated the findings from the embryos.

Curcumin affords protection against valproic acid induced teratogenicity by curtailing oxidative stress and inhibiting CYP2C9 activity

Administration of drugs during pregnancy is always done with immense caution, however multiple neurological ailments including epilepsy and depression warrant medical treatments even during pregnancy. This exposes the unborn fetus to killer teratogenic effects, thus warranting intense studies towards finding potential anti-teratogenic agents. We employed a valproic acid (VPA) induced model of fetotoxicity and teratogenicity in rats towards assessing the antiteratogenic activity of curcumin, an antioxidant well known for attenuating oxidative stress by increasing the content of glutathione and reducing the level of lipid hydroperoxide. We studied the level of GSH, catalase, SOD, ROS, TBARS and the activities of CYP2C9 and determined that VPA at a dose of 300 mg kg−1 body wt significantly decreased the levels of GSH, SOD and catalase and increased the levels of ROS, TBARS, mRNA expression and the levels of the CYP2C9 enzyme which is involved in the formation of the toxic metabolite (E)-2,4-diene-VPA. Upon co-administration of curcumin (100u2006150 and 200 mg kg−1 body wt) along with VPA the levels of GSH, SOD and catalase exhibited a significant increase and ROS, TBARS, mRNA expression and the level of CYP2C9 enzyme were found to be significantly decreased with respect to VPA. We conclude that the toxic metabolite (E)-2,4-diene-VPA is involved in the generation of oxidative stress subsequently contributing in the induction of malformations and anomalies and that curcumin affords dose dependent amelioration of the anomalies exerted by VPA. Our studies are suggestive of the fact that curcumin has antioxidant activity and can curtail the formation of toxic (E)-2,4-diene-VPA by inhibiting the CYP2C9 enzyme and finally protecting fetuses in a dose dependent manner.