Maternal haemoglobin and perinatal outcome in a tertiary care hospital in Jammu city, India

Background: Maternal anemia is a common problem worldwide. The objective of this study was to assess the prevalence of anemia in women attending a tertiary care centre in their third trimester of pregnancy and to study the relation of maternal hemoglobin with perinatal outcome.Methods: This study was conducted in the Department of Obstetrics and Gynecology, SMGS Hospital, Jammu for a period of one year from Nov. 2015 to Oct. 2016. A total of 767 women in their third trimester (>28 weeks) were enrolled and were followed till delivery. Their hemoglobin levels were noted and its association with adverse maternal and fetal outcomes was studied.Results: It was seen that out of 767 patients, 193 (25.16%) were non-anemic and 574 (74.83%) were anemic out of which 343 (44.71%) were mildly anemic, 160 (20.86%) had moderate anemia and 70 (9.12%) had severe anemia. There was increased incidence of preterm delivery, PPH, preeclampsia, eclampsia, CCF, mortality & blood transfusions in the anemic group as compared to the non-anemic group. Among the adverse fetal outcomes, there was increased incidence of intrauterine deaths, intrauterine growth restriction, NICU admissions, meconium stained liquor, low birth weight babies and APGAR scores <7/10 among the anemic group as compared to the non-anemic group.Conclusions: Maternal anemia is a significant risk factor for adverse maternal and perinatal outcome and its prevalence continues to be high especially in developing countries like India. So, correction of maternal anemia still remains one of the most important step towards better obstetrical care

Cell-based screening of antistress activity of some phytochemicals: Identification, validation, and relevance to old-age related pathologies

Background: A variety of environmental stresses have been shown to contribute to poor quality of life, tissue dysfunctions and ailments including metabolic disorders, cognitive impairment, and accelerated aging. Oxidative stress (an imbalance between the production and processing of highly reactive oxygen species) is largely associated with these phenotypes. Whereas drug development and disease therapeutics have advanced remarkably in last three decades, there are still limited options for stress management. Since the later can effectively decrease the disease burden, we aimed to screen phytochemicals with anti-oxidative stress activity using cell-based assays. Methods: Brain-derived cells were subjected to chemical models of oxidative (paraquat), metal (cadmium nitrate) or hypoxia (cobalt chloride) stresses. Stressed cells were allowed to recover either in the control or phytochemical supplemented culture medium. Cell survival and protein expression/signaling were analyzed to select the useful compounds and/or plant extracts. Results: Cells subjected to paraquat stress showed decrease in their viability. Three rounds of blind screening of the 24 phytochemicals resulted in identification of 5 compounds that caused better recovery of cells. The selected compounds were examined for their ability for protection against metal and hypoxia stresses induced by cadmium nitrate and cobalt chloride, respectively. Based on these 3D-anti stress protection ability, Withanone (Wi-N) and triethylene glycol (TEG) were selected for molecular validation. We found that whereas stress caused increase in (i) apoptosis (ii) ROS accumulation coupled with mitochondrial depolarization (iii) DNA double-strand break (iiii) protein aggregation, the selected compounds and the Ashwagandha extracts (known to possess these compounds) caused remarkable protection. Furthermore, both Wi-N and TEG caused differentiation of C6 glioblastoma and IMR-32 neuroblastoma as evidenced by the respective differentiated cell morphology and increased expression of biomarkers. Conclusion: The results suggested that Wi-N, TEG, their mixture, and the natural resource (Ashwagandha) possess potent antistress activity that may be useful for management of old-age-related ailments

Identification of Tectorigenin as a natural pro-hypoxia compound: implications in modulation of cellular differentiation and senescence

Background: Hypoxia, a suboptimal level of oxygen, evokes stress response in cells and activated hypoxia signaling has been largely established as a pro-metastasis and pro-angiogenic factor for tumor cells. On the other hand, age-related neurodegenerative disorders are characterized by hypoxic environment, accumulation of molecular garbage and induction of premature senescence. Several recent studies have reported anti-stress impact of the intermittent induction of hypoxia signaling in these cells. Methods: Screening of a phytochemical library using Hypoxia Responsive Element (HRE) driven luciferase as a reporter was carried out to identify hypoxia-modulating phytochemicals. Activation of HIF-1a (master regulator of hypoxia signaling) was validated by Western Blotting and immunostaining using specific antibodies. Short-term and long-term effect of the selected compounds on cell viability were determined by cell viability and colony forming assays, respectively. Furthermore, in vitro wound-scratch assays, protein aggregation models, and replicative senescence models were recruited to determine the effect of the selected compound on these phenotypes. Results: Tectorigenin (TEC) (iso-flavone obtained from leopard lily or Iris domestica) was selected as a pro-hypoxia factor. TEC treated cells showed significant activation HRE-driven luciferase reporter and upregulation of endogenous HIF-1a. On these lines, it was found that TEC resulted in de-aggregation of induced aggregation of protein reporters. cDNA microarray data revealed that TEC modulated the expression of genes involved in cell migration and differentiation. We used cellular senescence and astrocytic differentiation models and found lifespan extension of normal human fibroblasts and differentiation of rat-glioma cells, respectively. Conclusions: TEC could be defined as an anti-stress and anti-aging phytochemical that could be useful to manage hypoxia-driven ailments, involving protein aggregation and neurodegeneration. Further studies are warranted to support these claims and to dissect their molecular mechanism(s) of action

Inhibition of lipid accumulation in hepatocytes by unique Ashwagandha extracts

Introduction: Ashwaganda (Withania Somnifera) is a popular ayurvedic herb, trusted for a variety of health benefits in Indian traditional home medicine system. Steroidal lactones, Withaferin A (Wi-A) and Withanone (Wi-N), have been characterized as its major bioactives with a variety of bioactivities. We investigated the effect of Ashwagandha extracts on steatosis, abnormal retention of fat within a cell or organ that often affects liver as non-alcoholic fatty liver disease (NAFLD). Methods: We prepared extracts from Ashwagandha that varied in their Wi-A and Wi-N content. Cytotoxicity of these extracts on human hepatocytes (Huh-7 and Suit-2) was evaluated by cell viability assays. Nontoxic doses were used to treat the cells subjected to activated lipid accumulation by palmitic acid (PA). The lipolygenesis was evaluated by Oil Red O and triglyceride (TG) assays, and the expression of molecules involved in this process. Results and Discussions: The four kinds of extracts with different amounts of total withanolides and Wi-A:Wi-N ratio were generated. Cells were treated with PA to induce lipid accumulation. We found that in cells pre-treated with specific Ashwahandha extracts, TG accumulation was decreased. Of note, Sterol regulatory element-binding protein-1c (SREBP-1c), and its downstream effector-Fas, the key regulators of lipogenesis showed downregulation in specific extract-treated cells. Furthermore, the expression of PPARγ, a key factor involved in hepatic lipogenesis, showed decrease in cells treated with some of these extracts. Conclusion: Ashwagandha extracts may provide a useful natural resource with anti-steatosis activity, maintaining liver health and NAFLD prevention

Molecular insights to the dose-dependent activities of Ashwagandha extracts

Background: Stress is an inevitable component of life. Several herbs are known for their health supporting effects that range from treatment of stress, common cold to cancer. We investigated the dose-dependent effect of Ashwagandha (Withania somnifera) extracts on human normal and cancer cells, and have attempted to resolve the molecular mechanisms of their antistress activities. Methods: Ashwagandha extracts were chemically profiled by HPLC. Cytotoxicity was determined by viability assays. Biochemical and immunoimaging assays were performed using specific antibodies. Results: Human normal cells treated with low doses of the leaf extract or purified withanolides (Withaferin A or Withanone) showed no toxicity. Such non-toxic doses were selected for antistress, neurodifferentiation and neuroregenerative assays. We found that whereas normal cells exposed to oxidative and UV stresses showed poor viability/growth arrest/apoptosis, cells treated with low doses of Ashwagandha extracts were protected. Brain-derived cells exposed to glutamate and scopolamine stresses showed protection and strong differentiation as marked by expression of neurodifferentiation markers. Muscle-derived cells cultured in low doses of extract showed muscle differentiation as marked by expression of muscle differentiation markers. Most recently, using computational tools, we examined potential of Ashwagandha for anti-SARS-CoV2 virus activity, and found that most of the Ashwagandha Withanolides have potential to block cell surface receptors (ACE2 and TMPRSS2) that are involved in entry of virus to human cells. Furthermore, Ashwagandha treated cells showed decrease in ACE2 and TMPRSS2 expression suggesting its potential in blocking virus infection. Conclusion: Ashwagandha extracts and withanolides possess useful bioactivities

Folate receptor mediated targeting enhances selective cytotoxicity of Ashwagandha derived drugs to cancer cells

Background: Folate receptors (FRs) have been shown to be overexpressed on the surface of a variety of cancer cells and their expression are limited in normal cells and tissues. Since FR strongly binds to folic acid (FA), FA-functionalized nanocarriers have been proposed as a reliable strategy for delivery of anticancer drugs. We have earlier reported that the alcoholic extract of Ashwagandha leaves (i-Extract) and its major cytotoxic component, Withaferin A (Wi-A), have cancer cell killing activity. In the present study, we synthesized a FR-targeting i-Extract nanocomplex (FRi-ExNC) and a FR-targeting Wi-A nanocomposite (FRWi-ANC), by conjugating FA to polyethylene glycol and amphiphilic nanoframeworks, respectively. We investigated their anticancer potentials in in vitro and in vivo assays. Methods: Selective cellular uptake of FRi-ExNC and FRWi-ANC were evaluated by immunofluorescent microscopy. Cytotoxic effect of FRi-ExNC and FRWi-ANC in cancer cells were detected by assays including cell viability, apoptosis and biochemical determination of proteins involved in these phenotypes. The antitumor efficacy of FRi-ExNC and FRWi-ANC were investigated by in vivo tumor formation assays in nude mice. Results: We found that FRi-ExNC and FRWi-ANC caused stronger cytotoxicity as seen by induction of apoptosis. It was confirmed by cell cycle and protein expression analyses. In vivo tumor growth assays for subcutaneous xenografts in nude mice also revealed significantly enhanced suppression of tumor growth in the treated groups. Conclusions: Our results suggested that these two kinds of nanoparticles serve as useful nanomedical tools for selective targeting of drugs to the cancer cells and enhanced anticancer activity

Molecular insights to the p53-independent anticancer activity of MortaparibMild

Background: Mortalin, a member of the Hsp70 family of proteins, enriched in many types of cancers, has been shown to promote carcinogenesis and metastasis by multiple ways of which inactivation of p53 has been firmly established. Downregulation of mortalin by small RNAs and disruption of mortalin-p53 interactions by small molecules have earlier been shown to activate tumor suppressor activities of p53 yielding growth arrest/apoptosis in cancer cells. In this premise, screening of a chemical library to identify potential abrogators of mortalin-p53 interaction was performed using two-way imaging assay involving (i) shift of mortalin staining pattern from perinuclear (characteristics of cancer cells) to pancytoplasmic (characteristics of normal cells) and (ii) nuclear enrichment of p53. Using these assays in four rounds of screenings of a chemical library, three compounds Mortaparib, MortaparibPlus and Mortalparibmild were identified [1-4]. They showed similar structure and caused inactivation of mortalin and PARP1 activities. Methods: In the current study, we investigated response of cancer cells, varying in their p53 status (wild, mutant or null), to mortaparibs. Molecular signalings involved in cell proliferation, growth arrest, apoptosis, was determined by biochemical, imaging and expression analyses in control and Mortaparib (Mortaparibmild (4-[(4-amino-5-thiophen-2-yl-1,2,4-triazol-3-yl)sulfanylmethyl]-N-(4-methoxyphenyl)-1,3-thiazol-2-amine, in particular) treated cells. Results: We report that MortaparibMild, although with relatively weaker efficacy as compared to MortaparibandMortaparibplus, cause growth arrest or apoptosis in cancer cells by downregulation of mortalin and PARP-1 activities in p53-dependent and -independent pathways. Low nontoxic concentrations of the three Mortaparibs caused inhibition of cell migration, invasion and clusterization that was supported by molecular markers involved in these characteristics of cancer cells. Of note, in these assays, MortaparibMild showed good efficacy. These data suggested the three Mortaparibs possess differential activities that may be recruited for treatment of different stages of cancer targeting proliferation, metastasis and relapse of cancer. Conclusions: Taken together, we report anti-cancer activity of three mortaparibs (MortaparibMild in particular)and their differential use in cancer management that warrants further attention in laboratory and clinical studies

Molecular insights into the anti-stress function of fucoxanthin, a major component of an edible seaweed (wakame)

Background: Mortalin, a member of the Hsp70 family of proteins, enriched in many types of cancers, has been shown to promote carcinogenesis and metastasis by multiple ways of which inactivation of p53 has been firmly established. Downregulation of mortalin by small RNAs and disruption of mortalin-p53 interactions by small molecules have earlier been shown to activate tumor suppressor activities of p53 yielding growth arrest/apoptosis in cancer cells. In this premise, screening of a chemical library to identify potential abrogators of mortalin-p53 interaction was performed using two-way imaging assay involving (i) shift of mortalin staining pattern from perinuclear (characteristics of cancer cells) to pancytoplasmic (characteristics of normal cells) and (ii) nuclear enrichment of p53. Using these assays in four rounds of screenings of a chemical library, three compounds Mortaparib, MortaparibPlus and Mortalparibmild were identified [1-4]. They showed similar structure and caused inactivation of mortalin and PARP1 activities. Methods: In the current study, we investigated response of cancer cells, varying in their p53 status (wild, mutant or null), to mortaparibs. Molecular signalings involved in cell proliferation, growth arrest, apoptosis, was determined by biochemical, imaging and expression analyses in control and Mortaparib (Mortaparibmild (4-[(4-amino-5-thiophen-2-yl-1,2,4-triazol-3-yl)sulfanylmethyl]-N-(4-methoxyphenyl)-1,3-thiazol-2-amine, in particular) treated cells. Results: We report that MortaparibMild, although with relatively weaker efficacy as compared to MortaparibandMortaparibplus, cause growth arrest or apoptosis in cancer cells by downregulation of mortalin and PARP-1 activities in p53-dependent and -independent pathways. Low nontoxic concentrations of the three Mortaparibs caused inhibition of cell migration, invasion and clusterization that was supported by molecular markers involved in these characteristics of cancer cells. Of note, in these assays, MortaparibMild showed good efficacy. These data suggested the three Mortaparibs possess differential activities that may be recruited for treatment of different stages of cancer targeting proliferation, metastasis and relapse of cancer. Conclusions: Taken together, we report anti-cancer activity of three mortaparibs (MortaparibMild in particular)and their differential use in cancer management that warrants further attention in laboratory and clinical studies

Merger of Ayurveda and Tissue Culture-Based Functional Genomics: Inspirations from Systems Biology

Ayurveda is one of the ancient systems of health care of Indian origin. Roughly translated into "Knowledge of life", it is based on the use of natural herbs and herb products for therapeutic measures to boost physical, mental, social and spiritual harmony and improve quality of life. Although sheltered with long history and high trust, ayurveda principles have not entered laboratories and only a handful of studies have identified pure components and molecular pathways for its life-enhancing effects. In the post-genomic era, genome-wide functional screenings for targets for diseases is the most recent and practical approach. We illustrate here the merger of ayurveda and functional genomics in a systems biology scenario that reveals the pathway analysis of crude and active components and inspire ayurveda practice for health benefits, disease prevention and therapeutics

Cell-based experimental evidence to the anti-COVID-19 potential of Ashwagandha and honeybee propolis ingredients

Background: The COVID-19 pandemic emerged in December 2019 by a novel strain of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) has led to new endeavours in repurposing of existing drugs, anti-COVID-19 vaccine and drug development. Natural products, due to their general safety and wider availability, have attracted research and public attention. In this study, we report anti-COVID potential of compounds from honeybee propolis and Ayurvedic herb, Ashwagandha. Effect of active ingredients was studied on human cell surface receptors (ACE-2:Angiotensin Converting Enzyme-2/Spike protein and TMPRSS2:Transmembrane Protease Serine 2), critical for virus infection and virus main protease (Mpro, essential for virus replication), through molecular simulations and in vitro experiments. Methods: Structure-based computational analyses were performed to predict the effect of honeybee propolis (CAPE: Caffeic Acid Phenethyl Ester and ARC: Artepillin C), and Ashwagandha (Withanolides) ingredients on virus-host cell surface receptors. Cell-based assays were used to investigate the effect of these compounds on the expression level of the target proteins and virus replication. Results: Ashwagandha-derived nine withanolides were tested in silico for their potential to target and inhibit (i) ACE-2 and TMPRSS2 receptors (ii) viral main protease Mpro. We found that most withanolides possess capacity to bind to ACE-2, TMPRSS2 and Mpro . On the other hand, CAPE and ARC showed stable interactions at the active site of ACE2 and Mpro . ARC, but not CAPE, showed stable interaction with TMPRSS2. Human cells treated with withanolides, CAPE or ARC showed downregulation of both the receptors. Furthermore, celland PCR-based SARS-CoV-2 replication assays endorsed their antiviral activity. Conclusion: The findings suggest that the Ashwagandha-withanolides and honeybee propolis-derived compounds, CAPE, and ARC, could be helpful in the reduction of viral replication/infection, and hence warrant further experimental and clinical attention