The current study investigated transcriptional distortion in prostate cancer cells using the vitamin D receptor (VDR) as a tool to examine how epigenetic events driven by corepressor binding and CpG methylation lead to aberrant gene expression. These relationships were investigated in the non-malignant RWPE-1 cells that were 1α,25(OH)2D3 responsive (RWPE-1) and malignant cell lines that were 1α,25(OH)2D3 partially responsive (RWPE-2) and resistant (PC-3). These studies revealed that selective attenuation and repression of VDR transcriptional responses in the cancer cell lines reflected their loss of antiproliferative sensitivity. This was evident in VDR target genes including VDR, CDKN1A (encodes p21(waf1/cip1)) and GADD45A; NCOR1 knockdown alleviated this malignant transrepression. ChIP assays in RWPE-1 and PC-3 cells revealed that transrepression of CDKN1A was associated with increased NCOR1 enrichment in response to 1α,25(OH)2D3 treatment. These findings supported the concept that retained and increased NCOR1 binding, associated with loss of H3K9ac and increased H3K9me2, may act as a beacon for the initiation and recruitment of DNA methylation. Overexpressed histone methyltransferases (KMTs) were detectable in a wide panel of prostate cancer cell lines compared with RWPE-1 and suggested that generation of H3K9me2 states would be favored. Cotreatment of cells with the KMT inhibitor, chaetocin, increased 1α,25(OH)2D3-mediated induction of CDKN1A expression supporting a role for this event to disrupt CDKN1A regulation. Parallel surveys in PC-3 cells of CpG methylation around the VDR binding regions on CDKN1A revealed altered basal and VDR-regulated DNA methylation patterns that overlapped with VDR-induced recruitment of NCOR1 and gene transrepression. Taken together, these findings suggest that sustained corepressor interactions with nuclear-resident transcription factors may inappropriately transform transient-repressive histone states into more stable and repressive DNA methylation events.
Addressing inequitable coverage of maternal and child health care services among different socioeconomic strata of population and across states is an important part of India's contemporary health program. This has wide implications for the achievement of the Millennium Development Goal targets.
This paper assesses the inequity in coverage of maternal, newborn, and child health (MNCH) care services across household wealth quintiles in India and its states.
Utilizing the District Level Household and Facility Survey conducted during 2007–08, this paper has constructed a Composite Coverage Index (CCI) in MNCH care.
The mean overall coverage of 45% was estimated at the national level, ranging from 31% for the poorest to 60% for the wealthiest quintile. Moreover, a massive state-wise difference across wealth quintiles was observed in the mean overall CCI. Almost half of the Indian states and union territories recorded a ≤50% coverage in MNCH care services, which demands special attention.
India needs focused efforts to address the inequity in coverage of health care services by recognising or defining underserved people and pursuing well-planned time-oriented health programs committed to ameliorate the present state of MNCH care.
maternal; newborn and child health; composite coverage index; household economic status; states; India; millennium development goals
Although child immunization is regarded as a highly cost-effective lifesaver, about fifty percent of the eligible children aged 12–23 months in India are without essential immunization coverage. Despite several programmatic initiatives, urban-rural and gender difference in child immunization pose an intimidating challenge to India’s public health agenda. This study assesses the urban-rural and gender difference in child immunization coverage during 1992–2006 across six major geographical regions in India.
Data and Methods
Three rounds of the National Family Health Survey (NFHS) conducted during 1992–93, 1998–99 and 2005–06 were analyzed. Bivariate analyses, urban-rural and gender inequality ratios, and the multivariate-pooled logistic regression model were applied to examine the trends and patterns of inequalities over time.
The analysis of change over one and half decades (1992–2006) shows considerable variations in child immunization coverage across six geographical regions in India. Despite a decline in urban-rural and gender differences over time, children residing in rural areas and girls remained disadvantaged. Moreover, northeast, west and south regions, which had the lowest gender inequality in 1992 observed an increase in gender difference over time. Similarly, urban-rural inequality increased in the west region during 1992–2006.
This study suggests periodic evaluation of the health care system is vital to assess the between and within group difference beyond average improvement. It is essential to integrate strong immunization systems with broad health systems and coordinate with other primary health care delivery programs to augment immunization coverage.
Development of a vaccine to prevent or reduce parasite development in lymphatic filariasis would be a complementary approach to existing chemotherapeutic tools. Trehalose-6-phosphate phosphatase of Brugia malayi (Bm-TPP) represents an attractive vaccine target due to its absence in mammals, prevalence in the major life stages of the parasite and immunoreactivity with human bancroftian antibodies, especially from endemic normal subjects. We have recently reported on the cloning, expression, purification and biochemical characterization of this vital enzyme of B. malayi. In the present study, immunoprophylactic evaluation of Bm-TPP was carried out against B. malayi larval challenge in a susceptible host Mastomys coucha and the protective ability of the recombinant protein was evaluated by observing the adverse effects on microfilarial density and adult worm establishment. Immunization caused 78.4% decrease in microfilaremia and 71.04% reduction in the adult worm establishment along with sterilization of 70.06% of the recovered live females. The recombinant protein elicited a mixed Th1/Th2 type of protective immune response as evidenced by the generation of both pro- and anti-inflammatory cytokines IL-2, IFN-γ, TNF-α, IL-4 and an increased production of antibody isotypes IgG1, IgG2a, IgG2b and IgA. Thus immunization with Bm-TPP conferred considerable protection against B. malayi establishment by engendering a long-lasting effective immune response and therefore emerges as a potential vaccine candidate against lymphatic filariasis (LF).
Epigenetic modifiers play important roles in fine-tuning the cellular transcriptome. Any imbalance in these processes may lead to abnormal transcriptional activity and thus result in disease state. Distortions of the epigenome have been reported in cancer initiation and progression. DNA methylation and histone modifications are principle components of this epigenome, but more recently it has become clear that microRNAs (miRNAs) are another major component of the epigenome. Interactions of these components are apparent in prostate cancer (CaP), which is the most common non-cutaneous cancer and second leading cause of death from cancer in the USA. Changes in DNA methylation, altered histone modifications and miRNA expression are functionally associated with CaP initiation and progression. Various aspects of the epigenome have also been investigated as biomarkers for different stages of CaP detection, though with limited success. This review aims to summarize key aspects of these mechanistic interactions within the epigenome and to highlight their translational potential as functional biomarkers. To this end, exploration of TCGA prostate cancer data revealed that expression of key CaP miRNAs inversely associate with DNA methylation. Given the importance and prevalence of these epigenetic events in CaP biology it is timely to understand further how different epigenetic components interact and influence each other.
epigenetics; DNA methylation; histone modifications; microRNA; prostate cancer; cancer
India, with a population of more than 1.21 billion, has the highest maternal mortality in the world (estimated to be 56000 in 2010); and adolescent (aged 15–19) mortality shares 9% of total maternal deaths. Addressing the maternity care needs of adolescents may have considerable ramifications for achieving the Millennium Development Goal (MDG)–5. This paper assesses the socioeconomic differentials in accessing full antenatal care and professional attendance at delivery by adolescent mothers (aged 15–19) in India during 1990–2006.
Methods and Findings
Data from three rounds of the National Family Health Survey of India conducted during 1992–93, 1998–99, and 2005–06 were analyzed. The Cochran-Armitage and Chi-squared test for linear and non-linear time trends were applied, respectively, to understand the trend in the proportion of adolescent mothers utilizing select maternity care services during 1990–2006. Using pooled multivariate logistic regression models, the probability of select maternal healthcare utilization among women by key socioeconomic characteristics was appraised. After adjusting for potential socio-demographic and economic characteristics, the likelihood of adolescents accessing full antenatal care increased by only 4% from 1990 to 2006. However, the probability of adolescent women availing themselves of professional attendance at delivery increased by 79% during the same period. The study also highlights the desolate disparities in maternity care services among adolescents across the most and the least favoured groups.
Maternal care interventions in India need focused programs for rural, uneducated, poor adolescent women so that they can avail themselves of measures to delay child bearing, and for better antenatal consultation and delivery care in case of pregnancy. This study strongly advocates the promotion of a comprehensive ‘adolescent scheme’ along the lines of ‘Continuum of Maternal, Newborn and Child health Care’ to address the unmet need of reproductive and maternal healthcare services among adolescent women in India.
Sensing of microbial pathogens by pathogen-associated molecular patterns (PAMPs) through pattern recognition receptors (PRRs) elicits a defense program known as PAMP-triggered immunity (PTI). Recently, we have shown that the Arabidopsis thaliana L-TYPE LECTIN RECEPTOR KINASE-VI.2 (LecRK-VI.2) positively regulates bacterial PTI. In this report, we suggest by in silico analysis that the kinase domain of LecRK-VI.2 is functional. LecRK-VI.2 also demonstrated auto-phosphorylation activity in vitro in the presence of divalent metal cations indicating that LecRK-VI.2 has the ability to auto-phosphorylate. We further investigate the role of LecRK-VI.2 in Arabidopsis resistance to the necrotrophic fungal pathogen Botrytis cinerea. Disruption of LecRK-VI.2 did not affect Arabidopsis resistance to B. cinerea. Accordingly, wild-type upregulation levels of PTI-responsive WRKY53, FRK1, NHL10, CYP81F2 and CBP60 g after treatment with the fungal PAMP chitin were observed in lecrk-VI.2-1. These data provide evidences that the kinase domain of LecRK-VI.2 is active and show that LecRK-VI.2 is not critical for resistance to the fungal pathogen B. cinerea.
Arabidopsis thaliana; lectin receptor kinase; innate immunity; protein kinase; botrytis cinerea; chitin
Wegener's granulomatosis (WG) is a systemic necrotizing vasculitis that affects the small blood vessels. It mainly affects the upper and lower respiratory tract and kidneys. Central nervous system (CNS) involvement is rare, and has been reported only in about 8% of cases during the course of illness. Initial presentation with neurologic affection, particularly chronic hypertrophic meningitis is very unusual. We report the case of a 34 year old male who presented with chronic hypertrophic meningitis and multiple cranial nerve involvement as the initial manifestation, without respiratory and renal symptoms. This case highlights the difficulties in diagnosing a rare disease with rarer presentation, and at the same time illustrates that Wegener's granulomatosis should be considered in the differential diagnosis of chronic meningitis.
Chronic meningitis; cranial neuropathies; Wegener's granulomatosis
The phosphoglycerate mutase (PGM) enzyme catalyzes the interconversion of 2- and 3-phosphoglycerate in the glycolytic /gluconeogenic pathways that are present in the majority of cellular organisms. They can be classified as cofactor-dependent PGM (dPGM) or cofactor-independent PGM (iPGM). Vertebrates, yeasts, and many bacteria have only dPGM, while higher plants, nematodes, archaea, and many other bacteria have only iPGM. A small number of bacteria, including Escherichia coli and certain archaea and protozoa, contain both forms. The silencing of ipgm in Caenorhabditis elegans (C. elegans) has demonstrated the importance of this enzyme in parasite viability and, therefore, its potential as an anthelmintic drug target. In this study, the role of the Brugia malayi (B. malayi) ipgm in parasite viability, microfilaria release, embryogenesis, and in vivo development of infective larvae post-gene silencing was explored by applying ribonucleic acid (RNA) interference studies.
The in vitro ipgm gene silencing by small interfering RNA (siRNA) leads to severe phenotypic deformities in the intrauterine developmental stages of female worms with a drastic reduction (~90%) in the motility of adult parasites and a significantly reduced (80%) release of microfilariae (mf) by female worms in vitro. Almost half of the in vitro-treated infective L3 displayed sluggish movement. The in vivo survival and development of siRNA-treated infective larvae (L3) was investigated in the peritoneal cavity of jirds where a ~45% reduction in adult worm establishment was observed.
The findings clearly suggest that iPGM is essential for both larval and adult stages of B. malayi parasite and that it plays a pivotal role in female worm embryogenesis. The results thus validate the Bm-iPGM as a putative anti-filarial drug target.
siRNA; Lymphatic filariasis; Drug target; RNAi; Embryogenesis
Cholecalciferol (D3) supplementation results in variable increases in serum 25(OH)D3 levels, however, the influence of genetic polymorphisms on these variable responses is unclear. We measured serum 25(OH)D3, 24,25(OH)2D3, 1,25(OH)2D3 and VDBP levels in 50 colorectal cancer (CRC) patients before and during 2,000 IU daily oral D3 supplementation for six months and in 263 archived CRC serum samples. Serum PTH levels and PBMC 24-OHase activity were also measured during D3 supplementation. TagSNPs in CYP2R1, CYP27A1, CYP27B1, CYP24A1, VDR, and GC genes were genotyped in all patients, and the association between these SNPs and serum vitamin D3 metabolites levels before and after D3 supplementation was analyzed. The mean baseline serum 25(OH)D3 level was less than 32 ng/mL in 65 % of the 313 CRC patients. In the 50 patients receiving D3 supplementation, serum levels of 25(OH)D3 increased (p = 0.008), PTH decreased (p = 0.036) and 24,25(OH)2D3, 1,25(OH)2D3, VDBP levels and PBMC 24-OHase activity were unchanged. GC SNP rs222016 was associated with high 25(OH)D3 and 1,25(OH)2D3 levels at baseline while rs4588 and rs2282679 were associated with lower 25(OH)D3 and 1,25(OH)2D3 levels both before and after D3 supplementation. CYP2R1 rs12794714 and rs10500804 SNPs were significantly associated with low 25(OH)D3 levels after supplementation but not with baseline 25(OH)D3. Our results show that D3 supplementation increased 25(OH)D3 levels in all patients. GC rs4588 and rs2283679 SNPs were associated with increased risk of vitamin D3 insufficiency and suboptimal increase in 25(OH)D3 levels after D3 supplementation. Individuals with these genotypes may require higher D3 supplementation doses to achieve vitamin D3 sufficiency.
A key feature of innate immunity is the ability to recognize and respond to potential pathogens in a highly sensitive and specific manner. In plants, the first layer of defense is induced after recognition by pattern recognition receptors of microbe-associated molecular patterns. This recognition elicits a defense program known as pattern-triggered immunity. Pathogen entry into host tissue is a critical early step in causing infection. For foliar bacterial pathogens, natural surface openings such as stomata, are important entry sites. Stomata in contact with bacteria rapidly close and can thus restrict bacterial entry into leaves. The molecular mechanisms regulating stomatal closure upon pathogen perception are not yet well-understood. Plant lectin receptor kinases are thought to play crucial roles during development and in the adaptive response to various stresses. Although the function of most plant lectin receptor kinases is still not clear, a role for this kinase family in plant innate immunity is emerging. Here, we summarize recent progresses in the identification of lectin receptor kinases involved in plant innate immunity. We also discuss the role of lectin receptor kinases in stomatal innate immunity signaling.
plant; receptor-like kinase; lectin receptor kinase; innate immunity; stomatal innate immunity; bacteria
Apgar score (AS) is routinely used for assessment of newborns immediately after birth. Within acceptable limits, low saturations at birth are normal in vigorous newborn babies. Various studies have questioned the reliability of AS.
To detect whether AS is an accurate indicator of hypoxemia and to study the correlation of different components of AS with the arterial oxygenation saturation (SpO2) levels of normal newborns in the delivery room.
Settings and Design:
A prospective cross-sectional observational study on normal healthy neonates delivered vaginally in a tertiary level referral medical college.
Materials and Methods:
SpO2 levels were monitored continuously in the newborns with a pulse oximeter and serial recording of SpO2 levels was done at 5 min intervals starting at 1 min of life until 30 min after birth. Simultaneously, AS was recorded in these newborns at 1 and 5 min of life.
Data was analyzed using the Mann-Whitney–U test.
AS at 1 and 5 min of life didn’t correlate with the changes in SpO2 of newborns. In AS; though respiratory efforts and muscle tone were significantly correlated with SpO2 of the newborns, body color did not have significant correlation with simultaneously recorded SpO2.
A revised AS in which evaluation of color is replaced by pulse oximetry monitoring would prove to be a better tool for neonatal evaluation in the immediate postnatal period.
Apgar score; neonatal monitoring; oxygen saturation level; pulse oximetry
N-substituted pyridine hydrazide (pyridine-2-carbonyl chloride and 4-chloro-benzoic acid hydrazide) undergoes hydrazide formation of the iminic carbon nitrogen double bond through its reaction with cobalt(II), nickel(II), and copper(II) metal salts in ethanol which are reported and characterized based on elemental analyses, IR, solid reflectance, magnetic moment, molar conductance, and thermal analysis (TG). From the elemental analyses data, 1 : 2 metal complexes are formed having the general formulae [MCl2(HL)2] ·yH2O (where M = Co(II), Ni(II), and Cu(II), y = 1–3). The important infrared (IR) spectral bands corresponding to the active groups in the ligand and the solid complexes under investigation were studied. IR spectra show that ligand is coordinated to the metal ions in a neutral bidentate manner with ON donor sites. The solid complexes have been synthesized and studied by thermogravimetric analysis. All the metal chelates are found to be nonelectrolytes. From the magnetic and solid reflectance spectra, the complexes (cobalt(II), nickel(II), and copper(II)) have octahedral and square planner geometry, respectively. The antibacterial and antifungal activity's data show that the metal complexes have a promising biological activity comparable with the parent ligand against bacterial and fungal species.
Although the urban health issue has been of long-standing interest to public health researchers, majority of the studies have looked upon the urban poor and migrants as distinct subgroups. Another concern is, whether being poor and at the same time migrant leads to a double disadvantage in the utilization of maternal health services? This study aims to examine the trends and factors that affect safe delivery care utilization among the migrants and the poor in urban India.
Using data from the National Family Health Survey, 1992–93 and 2005–06, this study grouped the household wealth and migration status into four distinct categories poor-migrant, poor-non migrant, non poor-migrant, non poor-non migrant. Both chi-square test and binary logistic regression were performed to examine the influence of household wealth and migration status on safe delivery care utilization among women who had experienced a birth in the four years preceding the survey. Results suggest a decline in safe delivery care among poor-migrant women during 1992–2006. The present study identifies two distinct groups in terms of safe delivery care utilization in urban India – one for poor-migrant and one for non poor-non migrants. While poor-migrant women were most vulnerable, non poor-non migrant women were the highest users of safe delivery care.
This study reiterates the inequality that underlies the utilization of maternal healthcare services not only by the urban poor but also by poor-migrant women, who deserve special attention. The ongoing programmatic efforts under the National Urban Health Mission should start focusing on the poorest of the poor groups such as poor-migrant women. Importantly, there should be continuous evaluation to examine the progress among target groups within urban areas.
The trehalose metabolic enzymes have been considered as potential targets for drug or vaccine in several organisms such as Mycobacterium, plant nematodes, insects and fungi due to crucial role of sugar trehalose in embryogenesis, glucose uptake and protection from stress. Trehalose-6-phosphate phosphatase (TPP) is one of the enzymes of trehalose biosynthesis that has not been reported in mammals. Silencing of tpp gene in Caenorhabditis elegans revealed an indispensable functional role of TPP in nematodes.
Methodology and Principal Findings
In the present study, functional role of B. malayi tpp gene was investigated by siRNA mediated silencing which further validated this enzyme to be a putative antifilarial drug target. The silencing of tpp gene in adult female B. malayi brought about severe phenotypic deformities in the intrauterine stages such as distortion and embryonic development arrest. The motility of the parasites was significantly reduced and the microfilarial production as well as their in vitro release from the female worms was also drastically abridged. A majority of the microfilariae released in to the culture medium were found dead. B. malayi infective larvae which underwent tpp gene silencing showed 84.9% reduced adult worm establishment after inoculation into the peritoneal cavity of naïve jirds.
The present findings suggest that B. malayi TPP plays an important role in the female worm embryogenesis, infectivity of the larvae and parasite viability. TPP enzyme of B. malayi therefore has the potential to be exploited as an antifilarial drug target.
Lymphatic filariasis, one of the neglected tropical diseases, is the second leading cause of permanent and long term disability. Control of the disease relies on the mass administration of drugs which mainly act on the microfilariae without substantial effect on adult worms. Drugs need to be continued for several years to block the transmission of infection which may result in to development of resistant parasites. The sugar trehalose has been shown to play several important functions in the nematodes, and trehalose biosynthetic enzymes have been considered as potential targets for drug or vaccine candidate. In the present study we silenced trehalose-6-phosphate phosphatase and studied the biological function of TPP enzyme in the filarial nematode B. malayi viability, female worm embryogenesis and establishment of infection in the host. In vitro gene silencing was done in adult parasites using 5 mM concentration of siRNA while 2 mM of siRNA was used to treat L3 which were further inoculated into the peritoneal cavity of jirds to study the effect of siRNA treatment on in vivo larval development. The present findings validate trehalose-6-phosphate phosphatase as a vital antifilarial drug target.
Hodgkin lymphoma (HL) is a highly curable lymphoma with cure rates of over 80% and even higher with limited stage disease. Computerized tomography (CT) scan is currently the recommended modality in staging and assessment of response to therapy in patients with HL. However, CT has its limitations. This study describes our experience with patients of HL where fluorodeoxyglucose positron emission tomography (FDG-PET)-CT scan helped decide further management, after completion of chemotherapy.
This is a retrospective review of the records of children diagnosed with HL at our center. Patients with post-treatment CT scan showing evidence of residual disease, who underwent FDG-PET-CT for deciding further management, were included in the study.
Thirty one patients were diagnosed with HL during this period. Nine patients were eligible and underwent PET-CT. In 8 out of 9 patients, PET-CT showed no scan evidence of active disease. In one patient, FDG-PET-CT carried out after completion of chemotherapy showed evidence of active disease and was given radiotherapy.
FDG-PET-CT is a promising modality in deciding further management when there is discordance between the post-treatment CT scan and clinical condition of the patient with HL thus avoiding unnecessary chemotherapy/radiotherapy.
Children; hodgkin lymphoma; positron emission tomography-computerized tomography
This paper examines if, when controlling for biophysical and geographical variables (including rainfall, productivity of agricultural lands, topography/temperature, and market access through road networks), socioeconomic and health care indicators help to explain variations in the under-five mortality rate across districts from nine high focus states in India. The literature on this subject is inconclusive because the survey data, upon which most studies of child mortality rely, rarely include variables that measure these factors. This paper introduces these variables into an analysis of 284 districts from nine high focus states in India.
Information on the mortality indicator was accessed from the recently conducted Annual Health Survey of 2011 and other socioeconomic and geographic variables from Census 2011, District Level Household and Facility Survey (2007–08), Department of Economics and Statistics Divisions of the concerned states. Displaying high spatial dependence (spatial autocorrelation) in the mortality indicator (outcome variable) and its possible predictors used in the analysis, the paper uses the Spatial-Error Model in an effort to negate or reduce the spatial dependence in model parameters. The results evince that the coverage gap index (a mixed indicator of district wise coverage of reproductive and child health services), female literacy, urbanization, economic status, the number of newborn care provided in Primary Health Centers in the district transpired as significant correlates of under-five mortality in the nine high focus states in India. The study identifies three clusters with high under-five mortality rate including 30 districts, and advocates urgent attention.
Even after controlling the possible biophysical and geographical variables, the study reveals that the health program initiatives have a major role to play in reducing under-five mortality rate in the high focus states in India.
Family history and African-American race are important risk factors for both prostate cancer (CaP) incidence and aggressiveness. When studying complex diseases such as CaP that have a heritable component, chances of finding true disease susceptibility alleles can be increased by accounting for genetic ancestry within the population investigated. Race, ethnicity and ancestry were studied in a geographically diverse cohort of men with newly diagnosed CaP.
Individual ancestry (IA) was estimated in the population-based North Carolina and Louisiana Prostate Cancer Project (PCaP), a cohort of 2,106 incident CaP cases (2063 with complete ethnicity information) comprising roughly equal numbers of research subjects reporting as Black/African American (AA) or European American/Caucasian/Caucasian American/White (EA) from North Carolina or Louisiana. Mean genome wide individual ancestry estimates of percent African, European and Asian were obtained and tested for differences by state and ethnicity (Cajun and/or Creole and Hispanic/Latino) using multivariate analysis of variance models. Principal components (PC) were compared to assess differences in genetic composition by self-reported race and ethnicity between and within states.
Mean individual ancestries differed by state for self-reporting AA (p = 0.03) and EA (p = 0.001). This geographic difference attenuated for AAs who answered “no” to all ethnicity membership questions (non-ethnic research subjects; p = 0.78) but not EA research subjects, p = 0.002. Mean ancestry estimates of self-identified AA Louisiana research subjects for each ethnic group; Cajun only, Creole only and both Cajun and Creole differed significantly from self-identified non-ethnic AA Louisiana research subjects. These ethnicity differences were not seen in those who self-identified as EA.
Mean IA differed by race between states, elucidating a potential contributing factor to these differences in AA research participants: self-reported ethnicity. Accurately accounting for genetic admixture in this cohort is essential for future analyses of the genetic and environmental contributions to CaP.
Background & objectives:
Chapekar established a model of ovarian tumourigenesis in mice by splenic transplantation of ovaries, resulting in sustained luteinizing hormone (LH) levels because of absence of feedback inhibition. There is increasing evidence of the differential response to LH or hCG under various experimental conditions. The effect of sustained hormonal stimulation in long term cultures is sparsely investigated. The study is aimed to determine the role of hCG and LH stress on caprine ovarian granulosa cells and their downstream signaling in short and long term cultures.
To study the response of hCG and LH stress and downstream signaling, short term cultures were set up by exposing goat ovarian granulosa cells in primary cultures to hCG and LH stress (levels beyond their physiological doses) for 5 days (P0). Cells were sub-cultured at sixth day and subjected to prolonged LH/ hCG stress for two weeks in passage 1(P1) (long term cultures). Downstream cell signaling molecules were assessed. Intracellular cAMP was estimated by ELISA. For PKA and PKC, activity assays were performed. pERK protein expressions in short term cultures were assessed by Western blot and flowcytometry; in long term cultures, pERK expression was analyzed by flowcytometry.
Differential effects on cell proliferation were observed in long term cultures, where the untreated and hCG exposed cells showed markedly reduced cell proliferation after second week of exposure while LH treated cells continued to proliferate. Different levels of cAMP, PKA, PKC and phosphorylated ERK1/2 were observed on short term and long term LH stimulation. On sustained hormonal stimulation, cAMP levels were significantly (P<0.05) higher in hCG treated cultures as compared to controls and LH treated cultures. LH led to maximal elevation of ERK in long term cultures.
Interpretation & Conclusions:
As pERK1/2 promotes cellular proliferation, activation of ERK1/2 in LH treated cultures may be responsible for sustained growth. Prolonged LH treatment promoted growth and proliferation in caprine ovarian granulosa cells whereas prolonged exposure to hCG led to elevated levels of cAMP and decreased the rate of proliferation. Defining the signals and second messengers that act as survival or apoptotic mediators may help in elucidation of the mechanisms controlling proliferation or programmed cell death in granulosa cells.
Caprine; cell culture; cell proliferation; granulosa cells; hCG; LH
Coupled with the largest number of maternal deaths, adolescent pregnancy in India has received paramount importance due to early age at marriage and low contraceptive use. The factors associated with the utilization of maternal healthcare services among married adolescents in rural India are poorly discussed.
Using the data from third wave of National Family Health Survey (2005–06), available in public domain for the use by researchers, this paper examines the factors associated with the utilization of maternal healthcare services among married adolescent women (aged 15–19 years) in rural India. Three components of maternal healthcare service utilization were measured: full antenatal care, safe delivery, and postnatal care within 42 days of delivery for the women who gave births in the last five years preceding the survey. Considering the framework on causes of maternal mortality proposed by Thaddeus and Maine (1994), selected socioeconomic, demographic, and cultural factors influencing outcome events were included as the predictor variables. Bi-variate analyses including chi-square test to determine the difference in proportion, and logistic regression to understand the net effect of predictor variables on selected outcomes were applied. Findings indicate the significant differences in the use of selected maternal healthcare utilization by educational attainment, economic status and region of residence. Muslim women, and women belonged to Scheduled Castes, Scheduled Tribes, and Other Backward Classes are less likely to avail safe delivery services. Additionally, adolescent women from the southern region utilizing the highest maternal healthcare services than the other regions.
The present study documents several socioeconomic and cultural factors affecting the utilization of maternal healthcare services among rural adolescent women in India. The ongoing healthcare programs should start targeting household with married adolescent women belonging to poor and specific sub-groups of the population in rural areas to address the unmet need for maternal healthcare service utilization.
In this study we utilized the concept of rational drug design to identify novel compounds with optimal selectivity, efficacy and safety, which would bind to the target enzyme pteridine reductase 1 (PTR1) in Leishmania parasites. Twelve compounds afforded from Baylis-Hillman chemistry were docked by using the QUANTUM program into the active site of Leishmania donovani PTR1 homology model. The biological activity for these compounds was estimated in green fluorescent protein-transfected L. donovani promastigotes, and the most potential analogue was further investigated in intracellular amastigotes. Structure-activity relationship based on homology model drawn on our recombinant enzyme was substantiated by recombinant enzyme inhibition assay and growth of the cell culture. Flow cytometry results indicated that 7-(4-chlorobenzyl)-3-methyl-4-(4-trifluoromethyl-phenyl)-3,4,6,7,8,9-hexahydro-pyrimido[1,2-a]pyrimidin-2-one (compound 7) was 10 times more active on L. donovani amastigotes (50% inhibitory concentration [IC50] = 3 μM) than on promastigotes (IC50 = 29 μM). Compound 7 exhibited a Ki value of 0.72 μM in a recombinant enzyme inhibition assay. We discovered that novel pyrimido[1,2-a]pyrimidin-2-one systems generated from the allyl amines afforded from the Baylis-Hillman acetates could have potential as a valuable pharmacological tool against the neglected disease visceral leishmaniasis.
The overall survival of patients with acute myeloid leukemia (AML) remains poor due to both intrinsic and acquired chemotherapy resistance. Over expression of ATP binding cassette (ABC) proteins in AML cells has been suggested as a putative mechanism of drug resistance. Genetic variation among individuals affecting the expression or function of these proteins may contribute to inter-individual variation in treatment outcomes. DNA from pre-treatment bone marrow or blood samples from 261 patients age 20-85 years, who received cytarabine and anthracycline-based therapy at Roswell Park Cancer Institute between 1994 and 2006, was genotyped for eight non-synonymous single nucleotide polymorphisms in the ABCB1, ABCC1 and ABCG2 drug transporter genes. Heterozygous (AG) or homozygous (AA) variant genotypes for rs2231137 (G34A) in the ABCG2 (BRCP) gene, compared to the wild type (GG) genotype were associated with both significantly improved survival (HR=0.44, 95%CI=0.25-0.79), and increased odds for toxicity (OR=8.41, 95%CI=1.10-64.28). Thus genetic polymorphisms in the ABCG2 (BRCP) gene may contribute to differential survival outcomes and toxicities in AML patients via a mechanism of decreased drug efflux in both, AML cells and normal progenitors.
Acute myeloid leukemia; multidrug resistance; polymorphisms; survival; toxicity
The prevalence of irritable bowel syndrome (IBS) varies from 4% to 20% in different Asian nations. Prevalence of IBS in native North Indian community is not known.
Between November 2008 to December 2009, we estimated the prevalence of IBS in a rural community of Ballabgarh block, located in Haryana state. A structured questionnaire based on Rome III module was used to collect symptoms related to IBS from all the participants in a door to door survey. A Rome III criterion was used for diagnosis of IBS. IBS was further classified based on predominance of symptoms as constipation predominant, diarrhea predominant, mixed and unspecified based on Rome III module.
There were 4,767 participants (mean age 34.6 ± 10.8, males 50%). Overall, 555 (11.6%; 95% CI, 10.7-12.5) had constipation, 542 (11.4%; 95% CI, 10.5-12.3) diarrhea and 823 (17.3%; 95% CI, 16.2-18.4) abdominal pain. The overall prevalence of IBS was 4% (95% CI, 3.5-4.6). The prevalence of constipation predominant IBS was 0.3% (95% CI, 0.16-0.49), diarrhea predominant IBS 1.5% (95% CI, 1.18-1.90), mixed IBS 1.7% (95% CI, 1.35-2.11) and unsubtyped IBS 0.5% (95% CI, 0.32-0.75). The prevalence of IBS was significantly higher in females compared with males (4.8% vs 3.2%, P = 0.008). However, there was no significant difference between males and females in the prevalence of different subtypes of IBS. The prevalence increased with age.
The prevalence of IBS in a North Indian community is 4%. IBS poses a significant burden on the rural adults.
Asia; Constipation; Diarrhea; Epidemiology; Irritable bowel syndrome
The priming agent beta-aminobutyric acid (BABA) enhances Arabidopsis resistance to microbial pathogens and abiotic stresses through potentiation of the Arabidopsis defense responses. We have previously shown that BABA provokes a stress-induced morphogenic response, reduces vegetative growth and induces accumulation of anthocyanin. It was also found that L-Glutamine restores all tested BABA-induced phenotypes. Here we show that BABA induced transcripts accumulation of the two stress-responsive energy sensor protein kinases KIN10 and KIN11 and L-Glutamine inhibited this effect. It was also postulated that BABA induces a general amino acid stress response. BABA effect on Arabidopsis free amino acids content was thus analyzed. The amino acid balance was found to be altered by BABA treatment. Together these new data further suggest that BABA primes by stress imprinting.
arabidopsis; priming; SIMR; BABA; stress; imprinting; proline
The overall survival of patients with acute myeloid leukemia (AML) remains poor due to both intrinsic and acquired chemotherapy resistance. Over expression of ATP binding cassette (ABC) proteins in AML cells has been suggested as a putative mechanism of drug resistance. Genetic variation among individuals affecting the expression or function of these proteins may contribute to inter-individual variation in treatment outcomes. DNA from pre-treatment bone marrow or blood samples from 261 patients age 20-85 years, who received cytarabine and anthracycline-based therapy at Roswell Park Cancer Institute between 1994 and 2006, was genotyped for eight non-synonymous single nucleotide polymorphisms in the ABCB1, ABCC1 and ABCG2 drug transporter genes. Heterozygous (AG) or homozygous (AA) variant genotypes for rs2231137 (G34A) in the ABCG2 (BRCP) gene, compared to the wild type (GG) genotype were associated with both significantly improved survival (HR=0.44, 95%CI=0.25-0.79), and increased odds for toxicity (OR=8.41, 95%CI= 1.10-64.28). Thus genetic polymorphisms in the ABCG2 (BRCP) gene may contribute to differential survival outcomes and toxicities in AML patients via a mechanism of decreased drug efflux in both, AML cells and normal progenitors.
Acute myeloid leukemia; multidrug resistance; polymorphisms; survival; toxicity