Lead-210 content of food samples in India

Summary The paper presents results of measurements made on cereals and composite meal samples collected from Bombay market for their lead-210 content. The details of sampling and analytical chemistry procedures are also given. The Pb-210 contents of most of these samples were in the range of 1–5 pCi/kg of cereals samples. The concentrations in composite meal samples were mostly in the range of 1–3.5 pCi of²¹⁰Pb per composite meal. The assessment of daily intake of this isotope through food-stuffs has been made.     

An organelle proteomic method to study neurotransmission-related proteins, applied to a neurodevelopmental model of schizophrenia

Limited information is currently available on molecular events that underlie schizophrenia-like behaviors in animal models. Accordingly, we developed an organelle proteomic approach enabling the study of neurotransmission-related proteins in the prefrontal cortex (PFC) of postpubertal (postnatal day 60 (PD60)) neonatally ventral hippocampal (nVH) lesioned rats, an extensively used neurodevelopmental model of schizophrenia-like behaviors. The PFC was chosen because of its purported role in the etiology of the disease. Statistical analysis of 392 reproducible spots on 2-D organelle proteomic patterns revealed significant changes in intensity of 18 proteinous spots in plasma membrane-enriched fractions obtained from postpubertal nVH lesioned rats compared to controls. Mass spectrometric analysis and database searching allowed the identification of a single protein in each of the nine differential spots, including proteins of low abundance, such as neurocalcin delta. Most of the identified dysregulated proteins, including clathrin light chain B, syntaxin binding protein 1b and visinin-like protein 1 are known to be linked to various neurotransmitter systems and to play key roles in plasma membrane receptor expression and recycling as well as synaptic vesicle exocytosis/recycling. Organelle proteomic approaches have hence proved to be most useful to identify key proteins linked to a given behavior in animal models of brain diseases.     

Antigen-independent differentiation and maintenance of effector-like resident memory T cells in tissues

Differentiation and maintenance of recirculating effector memory CD8 T cells (T(EM)) depends on prolonged cognate Ag stimulation. Whether similar pathways of differentiation exist for recently identified tissue-resident effector memory T cells (T(RM)), which contribute to rapid local protection upon pathogen re-exposure, is unknown. Memory CD8αβ(+) T cells within small intestine epithelium are well-characterized examples of T(RM), and they maintain a long-lived effector-like phenotype that is highly suggestive of persistent Ag stimulation. This study sought to define the sources and requirements for prolonged Ag stimulation in programming this differentiation state, including local stimulation via cognate or cross-reactive Ags derived from pathogens, microbial flora, or dietary proteins. Contrary to expectations, we found that prolonged cognate Ag stimulation was dispensable for intestinal T(RM) ontogeny. In fact, chronic antigenic stimulation skewed differentiation away from the canonical intestinal T cell phenotype. Resident memory signatures, CD69 and CD103, were expressed in many nonlymphoid tissues including intestine, stomach, kidney, reproductive tract, pancreas, brain, heart, and salivary gland and could be driven by cytokines. Moreover, TGF-β-driven CD103 expression was required for T(RM) maintenance within intestinal epithelium in vivo. Thus, induction and maintenance of long-lived effector-like intestinal T(RM) differed from classic models of T(EM) ontogeny and were programmed through a novel location-dependent pathway that was required for the persistence of local immunological memory.     

Phytoextract-induced developmental deformities in malaria vector

Larvicidal potential of petroleum ether (Pee), carbon tetrachloride (Cte) and methanol extract (Mee) of Artemisia annua, Chenopodium album and Sonchus oleraceus was observed against malaria vector, Anopheles stephensi Liston. The Pee of A. annua with LC50 16.85 ppm after 24 h and 11.45 ppm after 48 h of treatment was found most effective, followed by Cte of A. annua and Ch. album, Pee of Ch. album and Mee of A. annua. However, no significant larvicidal activity was observed in Mee of Ch. album and all the three extracts of S. oleraceous. The Pee of A. annua was further investigated for its effect on the metamorphosis and the development of the malaria vector. It influenced the early life cycle of An. stephensi by reducing the percentage of hatching, larval, pupal and adult emergence and also lengthening the larval and pupal periods. The growth index was also reduced significantly. As the extract has remarkable effect on the metamorphosis and high larvicidal potential, it could, therefore, be used as an effective biocontrol agent against the highly nuisant malaria vector.     

Development of EST SSRs in Finger Millet (Eleusine coracana ssp coracana) and their Transferability to Pearl Millet (Pennisetum glaucum)

EST-SSR markers were developed using sequence information from 1740 expressed sequence tags (ESTs) of finger millet available in the public domain. A set of 31 SSR markers were synthesized based on di, tri, tetra and penta-nucleotide repeat sequences. These were used for PCR analysis of 11 elite germplasm lines of finger millet of Indian and African origin. Out of 31 SSR markers, amplification products were obtained for 17 primer pairs. Of these nine were found polymorphic with two alleles per locus. These 17 SSR primer pairs were also tested for amplification in three varieties of pearl millet (Pennisetum glaucum) and 11 could be transferred to pearl millet. The informative EST SSR markers developed, can be used in finger millet as well as pearl millet genetic improvement projects.     

Thioredoxin interacting protein (TXNIP) induces inflammation through chromatin modification in retinal capillary endothelial cells under diabetic conditions

Chronic hyperglycemia and activation of receptor for advanced glycation end products (RAGE) are known risk factors for microvascular disease development in diabetic retinopathy. Thioredoxin‐interacting protein (TXNIP), an endogenous inhibitor of antioxidant thioredoxin (TRX), plays a causative role in diabetes and its vascular complications. Herein we investigate whether HG and RAGE induce inflammation in rat retinal endothelial cells (EC) under diabetic conditions in culture through TXNIP activation and whether epigenetic mechanisms play a role in inflammatory gene expression. We show that RAGE activation by its ligand S100B or HG treatment of retinal EC induces the expression of TXNIP and inflammatory genes such as Cox2, VEGF‐A, and ICAM1. TXNIP silencing by siRNA impedes RAGE and HG effects while stable over‐expression of a cDNA for human TXNIP in EC elevates inflammation. p38 MAPK‐NF‐κB signaling pathway and histone H3 lysine (K) nine modifications are involved in TXNIP‐induced inflammation. Chromatin immunoprecipitation (ChIP) assays reveal that TXNIP over‐expression in EC abolishes H3K9 tri‐methylation, a marker for gene inactivation, and increases H3K9 acetylation, an indicator of gene induction, at proximal Cox2 promoter bearing the NF‐κB‐binding site. These findings have important implications toward understanding the molecular mechanisms of ocular inflammation and endothelial dysfunction in diabetic retinopathy. J. Cell. Physiol. 221: 262–272, 2009. © 2009 Wiley‐Liss, Inc     

Can human papillomavirus DNA testing of self-collected vaginal samples compare with physician-collected cervical samples and cytology for cervical cancer screening in developing countries?

Background: To determine human papillomavirus (HPV) types by polymerase chain reaction (PCR)-reverse line blot assay and examine the concordance between HPV by Hybrid Capture 2 (HC2) and PCR on self-collected vaginal and physician-collected cervical samples and cytology. Methods: This was a cross-sectional study of 546 sexually active women aged ≥30 years with persistent vaginal discharge, intermenstrual or postcoital bleeding or an unhealthy cervix. Participants self-collected vaginal samples (HPV-S) and physicians collected cervical samples for conventional Pap smear and HPV DNA (HPV-P) testing and performed colposcopy, with directed biopsy, if indicated. HPV testing and genotyping was done by HC2 and PCR reverse line blot assay. Concordance between HC2 and PCR results of self- and physician-collected samples was determined using a Kappa statistic (κ) and Chi-square test. Results: Complete data were available for 512 sets with 98% of women providing a satisfactory self-sample. PCR detected oncogenic HPV in 12.3% of self- and 13.0% of physician-collected samples. Overall, there was 93.8% agreement between physician-collected and self-samples (κ = 76.31%, 95% confidence interval [CI]: 64.97–82.29%, p = 0.04)—complete concordance in 473 cases (57 positive, 416 negative), partial concordance in seven pairs and discordance in 32 pairs. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of self-sampling for detection of cervical intraepithelial neoplasia (CIN)2+ disease were 82.5%, 93.6%, 52.4% and 98.4%, respectively; for physician-sampling they were 87.5%, 93.2%, 52.2% and 98.9%, respectively; and for cytology they were 77.5%, 87.3%, 34.1% and 97.9%, respectively. Concordance between HC2 and PCR was 90.9% for self-samples (κ = 63.7%, 95% CI: 55.2–72.2%) and 95.3% for physician-collected samples (κ = 80.4%, 95% CI: 71.8–89.0%). Conclusions: Self-HPV sampling compares favourably with physician-sampling and cytology. A rapid, affordable, HPV self-test kit can be used as the primary method of cervical cancer screening in low-resource situations.     

Prevalence of human papillomavirus infection among young women in North India

Background: The number of women infected with human papillomavirus (HPV) and the distribution of the HPV genotypes vary across populations and with age. Objective: To determine the prevalence and genotype distribution of HPV in young married women aged 16–24 years. Methods: 1300 women residing in an urban slum in Delhi donated samples of exfoliated cervical cells that were collected by the Digene^® kit and tested for the presence of HPV DNA by two techniques in parallel, i.e., PCR using PGMY consensus primers for all HPV types and the Digene HPV test (Hybrid Capture 2 (HC2) Probe B for high-risk (hr) types. Genotyping was done on all HPV positive samples using the Roche reverse line blot assay. Results: HPV infection was detected in 91/1300 (7%) samples by PCR and 110/1300 (8.4%) samples by HC2. Genotyping identified 20 high-risk and 11 low-risk types. HPV16 was the commonest high-risk type (3%) followed by HPV52 (1.2%) and HPV51 (0.8%). Among low-risk types, HPV62 was the commonest (0.8%), followed by HPV84 and HPV89 (0.5% each). Multiple infections were found in 3% of the HPV positive samples. Conclusion: A wide spectrum of HPV genotypes is seen in this young population. Knowledge about HPV types prevalent in communities in different regions of India would be useful in devising the optimum strategy for cervical cancer prevention.