Distribution, toxicity load and risk assessment of heavy metals in the groundwater of Dhemaji, Assam, India.

Metal contamination in drinking water has drawn attention since it gravely jeopardizes human health. This study was conducted in pre- and post-monsoon season in 2021 at Dhemaji, Assam, India. It characterized metal pollutants in groundwater, their distribution, possible sources, and evaluated the potential toxicity and associated health risk assessment. The seasonal mean concentration of Fe in both seasons is observed highest followed by Mn, Zn, Cu, As, and Ni. Furthermore, the metal concentrations during pre-monsoon are comparatively higher. The geogenic processes and agricultural practices are the major sources of groundwater metal contamination as evident from the statistical analysis. The different pollution indices viz. Heavy-metal Pollution Index (HPI), Heavy-metal Evaluation Index (HEI) and Degree of Contamination (Cd) suggested that groundwater is not suitable for drinking uses. The Heavy Metal Toxicity Load (HMTL) suggesting As, Co, Mn and Hg should be removed from the groundwater to ensure safety. Water pollution indices (WPI) suggest that Fe, Mn, As and Ni are the main pollution-causing metals in the study area which may be restored under the BIS and WHO limit by diluting the water. The human health risk has been calculated by carcinogenic and non-carcinogenic risk assessment. The non-carcinogenic risk for adults and children is within the threshold limit. The carcinogenic risk shows that continuous exposure of As and Ni may give rise to cancer among adults and children in the region. Therefore, comprehensive groundwater quality monitoring with well-planned treatment should be needed to provide safe and clean drinking water in the studied area.

Health information technology tools to accelerate gastrointestinal evaluation in patients with iron deficiency anaemia: a cluster randomised controlled trial.

OBJECTIVE: System-level safety measures do not exist to ensure that patients with iron deficiency anaemia (IDA) undergo proper diagnostic evaluations. We sought to determine if a set of EHR (electronic health record) tools and an expedited referral workflow increase short-term completion of bidirectional endoscopy in higher risk patients with IDA. MATERIALS AND METHODS: We conducted a pragmatic, cluster-randomised trial randomised by primary care physician (PCP) that included 16 PCPs and 316 patients with IDA. Physicians were randomised to intervention or control groups. Intervention components included a patient registry visible within the EHR, point-of-care alert and expedited diagnostic evaluation workflow for IDA. Outcomes were assessed at 120 days. The primary outcome was completion of bidirectional endoscopy. Secondary outcomes were any endoscopy completed or scheduled, gastroenterology consultation completed, and gastroenterology referral or endoscopy ordered or completed. RESULTS: There were no differences in the primary or secondary outcomes. At 120 days, the primary outcome had occurred for 7 (4%) of the intervention group and 5 (3.5%) of the control group. For the three secondary outcomes, rates were 15 (8.6%), 12 (6.9%) and 39 (22.4%) for the immediate intervention group and 10 (7.0%), 9 (6.3%) and 25 (17.6%) for the control group, respectively, p>0.2. Lack of physician time to use the registry tools was identified as a barrier. DISCUSSION AND CONCLUSION: Providing PCPs with lists of patients with IDA and a pathway for expedited evaluation did not increase rates of completing endoscopic evaluation in the short term. TRIAL REGISTRATION NUMBER: NCT05365308.

Characterization of Polycyclic Aromatic Hydrocarbons (PAHs) associated with fine aerosols in ambient atmosphere of high-altitude urban environment in Sikkim Himalaya.

Polycyclic Aromatic Hydrocarbons (PAHs) compounds are ubiquitous in ambient air due to their persistence, carcinogenicity, and mutagenicity. Gangtok being one of the cleanest cities in India located in Eastern Himalayan region, witnesses high developmental activities with enhanced urbanization affecting the ambient air quality. The present study aims to measure PM2.5 and PAHs in the ambient atmosphere of the Sikkim Himalaya to understand the influence of natural and anthropogenic activities on aerosol loading and their chemical characteristics. The PM2.5 samples were collected and analysed for the duration from Jan 2020 to Feb 2021.The seasonal mean concentrations of PM2.5 and PAHs were observed to be high during autumn and low during summer season. Overall, the annual mean concentration of PM2.5 was found higher than the prescribed limit of World Health Organization and National Ambient Air Quality Standards. The concentration of the 16 individual PAHs were found to be highest during autumn season (55.26 ± 37.15 ng/m3). Among the different PAHs, the annual mean concentration of fluorene (3.29 ± 4.07 ng/m3) and naphthalene (1.15 ± 3.76 ng/m3) were found to be the highest and lowest, respectively. The Molecular Diagnostic Ratio (MDR) test reveals higher contribution from heavy traffic activities throughout the winter and autumn seasons. The other possible sources identified over the region are fossil fuel combustion, and biomass burning. The multivariate statistical analysis (Multifactor Principal Component Analysis) also indicates a strong association between PM2.5 /PAHs and meteorological variables across the region in different seasons. The precipitation and wind pattern during the study period suggests that major contribution of the PM2.5 and PAHs were from local sources, with minimal contribution from long-range transport. The findings are important for comprehending the trends of PAH accumulation over a high-altitude urban area, and for developing sustainable air quality control methods in the Himalayan region.

COVID-19 and Poverty in South America: the Mental Health Implications.

The COVID-19 pandemic caused by the SARS-CoV-2 virus made governments across the world announce country-wide lockdowns suspending all activities and banning travels. This affected the lives of millions of people across all the continents in the world, including South America. The low-income classes were most affected by the lockdowns as many lost their only source of income. In addition, the uncertainty of the disease and its deadly nature along with lack of access to correct information led to a deterioration of the mental health of the low-income and indigenous groups in South America. The low-income population has little chance of gaining access to proper mental healthcare as many healthcare systems collapsed due to the nature of the pandemic. A good number of them faced short-term mental effects such as anxiety and panic attacks as well as long-term effects of depression. Proper steps must be taken to maintain and help the most vulnerable sections of the population. The pandemic along with the widespread poverty-stricken regions in the country have affected the population of the country. This article assesses the impact of COVID-19 on the poor and low-income population of the South American continent and the impact it has had on their mental health. It also reviews the recommended steps that can be taken to improve the situation and help the affected people.

Gut-specific arylphorin mediates midgut regenerative response against Cry-induced damage in Achaea janata.

Development of insect resistance to biopesticides is a current and pertinent global issue. Earlier, it was established that lepidopteran larvae can recover from Bt intoxication via a midgut regenerative response and subsequently generate resistance. Molecular aspects of restoration of the midgut integrity following toxin exposure are emerging recently. In the present study, we bring out the pivotal role of gut arylphorin in mediating the midgut regenerative response following sublethal Bt exposure in Achaea janata. Bt-induced midgut damage was characterized by microscopic analysis using differential interference contrast (DIC) and immunofluorescence (IF). Extensive disruption of brush-border membrane, associated with the underlying cytoskeletal alterations including F-actin, α-actin and ß-tubulin was observed. Single-photon fluorescence microscopy combined with fluorescence lifetime imaging (FLIM) established the metabolic state associated with enhanced stem cell proliferation and migration from the basal side towards the luminal side following the damage. In-silico analysis revealed the phylogenetic relationship of gut arylphorin with closely related insect species and indicated the presence of two different subunits. Transient RNAi knockdown of the arylphorin resulted in diminished expression of mitotic Cyclin B mRNA levels. Human monoclonal Cyclin B antibody cross-reactivity with the Cyclin B located in the stem cells further validate the role of arylphorin as the mitogenic factor responsible for stem cell proliferation and epithelial regeneration. An in-depth understanding of resistance mechanisms will aid in the design of new strategies for the long-term usage and efficacy of Bt technology against pest control.

Midgut aminopeptidase N expression profile in castor semilooper (Achaea janata) during sublethal Cry toxin exposure.

The midgut of lepidopteran larvae is a multifunctional tissue that performs roles in digestion, absorption, immunity, transmission of pathogens and interaction with ingested various molecules. The proteins localized at the inner apical brush border membrane are primarily digestive proteases, but some of them, like aminopeptidase N, alkaline phosphatase, cadherins, ABC transporter C2, etc., interact with Crystal (Cry) toxins produced by Bacillus thuringiensis (Bt). In the present study, aminopeptidase N (APN) was characterized as Cry-toxin-interacting protein in the larval midgut of castor semilooper, Achaea janata. Transcriptomic and proteomic analyses revealed the presence of multiple isoforms of APNs (APN1, 2, 4, 6 and 9) which have less than 40% sequence similarity but show the presence of characteristic 'GAMENEG' and zinc-binding motifs. Feeding a sublethal dose of Cry toxin caused differential expression of various APN isoform. Further, 6thgeneration Cry-toxin-exposed larvae showed reduced expression of APN2. This report suggests that A. janata larvae exploit altered expression of APNs to overcome the deleterious effects of Cry toxicity, which might facilitate toxin tolerance in the long run.

Chemical-free transfer of patterned reduced graphene oxide thin films for large area flexible electronics and nanoelectromechanical systems.

In this paper, a wet-dry hybrid technique to transfer patterned reduced graphene oxide (rGO) thin film to arbitrary substrates at predetermined locations without using any chemicals is reported. The transfer process involves water-assisted delamination of rGO, followed by dry transfer to an acceptor substrate using viscoelastic stamp. Patterned reduced graphene oxide films are transferred to silicon dioxide (SiO2/Si) substrate to begin with. Subsequently, the method is deployed to transfer rGO to different polymer substrates such as poly(methyl methacrylate) (PMMA), and crosslinked poly(4-vinylphenol) (c-PVP), which are commonly used as gate dielectric in flexible electronic applications. The credibility of the transfer process with precise spatial positioning on the target substrate leads to fabrication of freely suspended reduced graphene oxide membrane towards nanoelectromechanical systems (NEMS) based devices such as nanomechanical drum resonators.

Controlled release of sex steroids through osmotic pump alters brain GnRH1 and catecholaminergic system dimorphically in the catfish, Clarias gariepinus.

The present study aimed to evaluate osmotic pump-mediated controlled release of estrogen in males and androgen in females to analyze the impact on gonadotropin-releasing hormone (GnRH1), catecholamines (CAs) and other associated genes in the catfish, Clarias gariepinus. During pre-spawning phase, catfish were separately implanted osmotic pumps loaded with 17ß-estradiol (E2) in males and 17α-methyltestosterone (MT) in females at a dose of 10 µg/100 µl or saline (100 µl) controls into both sexes to release for 21 days and all fishes were maintained as per the duration. Further, GnRH1 expression levels were analysed in the discrete regions of brain after E2 and MT treatments in male and female catfish, respectively using qPCR which revealed that GnRH1 expression was significantly higher in E2 treated male as compared to the control. On the other hand, GnRH1 expression was lower in MT treated female when compared to the control in the discrete regions of brain. In addition, certain brain and monoaminergic system related genes showed a differential response. Catfish GnRH1 could be localized in preoptic area-hypothalamus (POA-HYP) that correlated with the expression profile in the discrete regions of catfish brain. Serum levels of sex steroids in the treated male fish indicated that the treatment of E2 could maintain and impart feminization effect even in the presence of endogenous androgen during gonadal recrudescence while such an effect was not seen in females with androgen treatment. Measurement of CAs, L-3,4-dihydroxyphenylalanine, dopamine and norepinephrine levels in the male and female brain after the controlled release of E2 and MT, respectively confirmed the modulation of neurotransmitters in the E2treated male than MT treated female fish. These results collectively suggest the severity of estrogenic over androgenic compounds to alter reproductive status even at a minimal dose by targeting CAs and GnRH1 at the level of brain of catfish. This study provides insights into the reproductive toxicity of sex steroid analogues at the level of brain GnRH1 and CA-ergic system in addition to serum T, 11-KT and E2 levels during gonadal recrudescence, which is a crucial period of gametogenesis preceding spawning.

The incisive papilla as a guide to maxillary anterior teeth position for complete dentures.

CONTEXT: The incisive papilla as a guide for complete dentures. AIMS: To evaluate incisive papilla as a guide to maxillary anterior teeth position for complete dentures. SETTINGS AND DESIGN: To measure the linear distances from the incisive papilla to the maxillary central incisors in Odia population and compare it to other ethnic groups. SUBJECTS AND METHODS: The subjects were selected following inclusion & exclusion criteria. The impressions of maxillary & mandibular arches were taken and poured with die stone to produce the cast. The incisive papilla & incisors were marked on casts with a pencil. The photographs of casts were traced on an acetate tracing paper. The anterior-most points of maxillary incisors, anterior, middle & posterior-most points of incisive papillae were marked on the tracing as A, I1,M, I2 respectively. The A to M and A to I2 distances were measured, analysed and compared with Caucasians, Southern Chinese, and Dravidians. STATISTICAL ANALYSIS: Independent sample "t" test and Analysis of Variance followed by the post-hoc Bonferroni test were used. RESULTS: A sample of 100 subjects in the age group of 20 to 40 years with was selected. The A to M measurements of Odia population differed significantly from Caucasians but were similar to Southern Chinese population. The A to I2 measurements of Odia population differed significantly from Caucasians & Dravidians but were similar to Southern Chinese. CONCLUSION: The linear measurements from incisive papilla to maxillary incisors of the Odia population are similar to that of Southern Chinese but differ significantly from Caucasians and Dravidians.

RNA-Seq analysis and de novo transcriptome assembly of Cry toxin susceptible and tolerant Achaea janata larvae.

Larvae of most lepidopteran insect species are known to be voracious feeders and important agricultural pests throughout the world. Achaea janata larvae cause serious damage to Ricinus communis (Castor) in India resulting in significant economic losses. Microbial insecticides based on crystalline (Cry) toxins of Bacillus thuringiensis (Bt) have been effective against the pest. Excessive and indiscriminate use of Bt-based biopesticides could be counter-productive and allow susceptible larvae to eventually develop resistance. Further, lack of adequate genome and transcriptome information for the pest limit our ability to determine the molecular mechanisms of altered physiological responses in Bt-exposed susceptible and tolerant insect strains. In order to facilitate biological, biochemical and molecular research of the pest species that would enable more efficient biocontrol, we report the midgut de novo transcriptome assembly and clustering of susceptible Cry toxin-exposed and Cry toxin tolerant Achaea janata larvae with appropriate age-matched and starvation controls.

Cutaneous Melanoma, Version 2.2019, NCCN Clinical Practice Guidelines in Oncology.

The NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) for Cutaneous melanoma have been significantly revised over the past few years in response to emerging data on immune checkpoint inhibitor therapies and BRAF-targeted therapy. This article summarizes the data and rationale supporting extensive changes to the recommendations for systemic therapy as adjuvant treatment of resected disease and as treatment of unresectable or distant metastatic disease.

Evidence for absence of bilateral transfer of olfactory learned information in Apis dorsata and Apis mellifera.

The capacity and condition under which the lateral transfer of olfactory memory is possible in insects is still debated. Here, we present evidence in two species of honeybees, Apis mellifera and Apis dorsata, consistent with the lack of ability to transfer olfactory associative memory in a proboscis extension response (PER) associative conditioning paradigm, where the untrained antenna is blocked by an insulating coat. We show that the olfactory system on each side of the bee can learn and retrieve information independently and the retrieval using the antenna on the side contralateral to the trained one is not affected by the training. Using the setup in which the memory on the contralateral side has been reported at 3 h after training, we see that the memory is available on the contralateral side immediately after training. In the same setup, coating the antenna with an insulator on the training side does not prevent learning, pointing to a possible insufficiency of the block of odor stimuli in this setup. Moreover, the behavior of the bee as a whole can be predicted if the sides are assumed to learn and store independently, and the organism as a whole is able to retrieve the memory if either of the sides have the memory.

Midgut de novo transcriptome analysis and gene expression profiling of Achaea janata larvae exposed with Bacillus thuringiensis (Bt)-based biopesticide formulation.

India is the major producer and exporter of castor oil in the world. Castor semilooper, Achaea janata is one of the main castor crop pests, which causes serious economic loss of crop, hence management and control of the pest are important. Currently, Bacillus thuringiensis (Bt) based biopesticides are being used for their control. However, the insects are known to develop resistance not only against chemical pesticides but also to Bt based biopesticides. In the present study, de novo transcriptome analysis was conducted to monitor the expression pattern of larval midgut genes in Achaea janata exposed to sublethal dose of Bt formulation. A total of 34,612 and 41,109 transcripts were identified in control and toxin-exposed larval midgut samples out of which 18,836 in control and 21,046 in toxin-exposed samples are annotated. Microarray data analysis employed to monitor the gene expression upon Cry toxin exposure revealed that 375 genes were upregulated and 579 genes were downregulated during all the time points (12-60 h) of toxin exposure. The differentially expressed transcripts include i.e. Cry toxin receptors, gut proteases, arylphorin, REPATs, detoxification enzymes and aquaporins. Validation of microarray data was performed by real-time quantitative PCR using few randomly selected genes and the results obtained were in corroboration. This is the first study on transcriptome data from the castor semilooper and the results would provide valuable resources for the characterization of Bt toxin response in the pest.

Purification and characterization of Bowman-Birk and Kunitz isoinhibitors from the seeds of Rhynchosia sublobata (Schumach.) Meikle, a wild relative of pigeonpea.

Rhynchosia sublobata, a wild relative of pigeonpea, possesses defensive proteinase/protease inhibitors (PIs). Characterization of trypsin specific PIs (RsPI) separated from seeds by column chromatography using 2-D gel electrophoresis and Edman degradation method identified R. sublobata possessed both Bowman-Birk isoinhibitors (RsBBI) and Kunitz isoinhibitors (RsKI). A quick method was developed to separate RsBBI and RsKI from RsPI based on their differential solubility in TCA and acetate buffer. N-terminus sequencing of RsBBI and RsKI by MALDI-ISD ascertained the presence of Bowman Birk and Kunitz type isoinhibitors in R. sublobata. RsBBI (9216 Da) and RsKI (19,412 Da) exhibited self-association pattern as revealed by western blotting with anti-BBI antibody and MALDI-TOF peptide mass fingerprint analysis, respectively. RsBBI and RsKI varied significantly in their biochemical, biophysical and insecticidal properties. RsBBI inhibited the activity of trypsin (Ki = 128.5 ±â€¯4.5 nM) and chymotrypsin (Ki = 807.8 ±â€¯23.7 nM) while RsKI (Ki = 172.0 ±â€¯9.2 nM) inhibited the activity of trypsin alone, by non-competitive mode. The trypsin inhibitor (TI) and chymotrypsin inhibitor (CI) activities of RsBBI were stable up to 100 °C. But, RsBBI completely lost its TI and CI activities on reduction with 3 mM DTT. Conversely, RsKI lost its TI activity on heating at 100 °C and retained >60% of its TI activity in presence of 3 mM DTT. CD spectroscopic studies on RsBBI and RsKI showed their secondary structural elements in the following order: random coils > ß-sheets/ß-turns > α-helix. However, RsKI showed reversible denaturation midpoint (Tm) of 75 °C. Further, the significant inhibitory activity of RsBBI (IC50 = 24 ng) and RsKI (IC50 = 59 ng) against trypsin-like gut proteases of Achaea janata (AjGPs) and Helicoverpa armigera (HaGPs) suggest them as potential biomolecules in the management of A. janata and H. armigera, respectively.

Report of a case of platelet-rich fibrin-mediated revascularization of immature 12 with histopathological evaluation.

The procedure of regeneration has shown promising results in the treatment of nonvital teeth with immature apices. However, the tissue being regenerated can only be confirmed through histological examination. This report describes an immature nonvital 12 with apical pathology which was treated via revascularization using 3% NaOCl and 17% ethylenediaminetetraacetic acid as irrigants; Ca(OH)2 as intracanal medicament and platelet-rich fibrin (PRF) as scaffold. On follow-up at 6 and 12 months, healing of periapical lesion, dentinal thickening, and apical closure with a canal exit forming distally were evident. The tooth, however, got fractured and was subsequently extracted at 12 months and sent for histopathological examination. It was revealed that a collagenous matrix for forming hard tissue similar to acellular cementum with a diffuse area of calcification could be seen in pulp space being attached with root canal dentin walls, suggesting that PRF may yield favorable outcomes in regenerative endodontic procedures.

Evaluation of anti-inflammatory and immunomodulatory effects of Premna integrifolia extracts and assay-guided isolation of a COX-2/5-LOX dual inhibitor.

Premna integrifolia (Agnimantha brihat) is a traditional medicinal plant with a prominent place in Ayurveda, Siddha and Unani systems of medicine. In this study we have evaluated the anti-inflammatory and immunomodulatory properties of the Premna integrifolia root extracts employing cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and 5-lipoxygenase (5-LOX) enzyme-based assays, lymphocyte proliferation assay, pro-and anti-inflammatory cytokines measurement. Petroleum ether extract (PEE) of Premna integrifolia showed potent inhibition of COX-2 and 5-LOX with IC50 values of 6.15 µg/mL and 11.33 µg/mL respectively. In in vitro studies on RAW 264.7 cell line, PEE showed inhibition in the formation of nitric oxide (NO), pro-inflammatory cytokines (IL-1ß, IL-6), prostaglandin E2 (PGE2) production, induction of anti-inflammatory cytokine (IL-2) and down-regulation of expression of COX-2, 5-LOX, TNF-α, IL-1ß and iNOS. PEE also significantly reduced carrageenan-induced paw edema in mouse model of inflammation. Further, attempts in isolating the active principle(s) involved in these anti-inflammatory effects of PEE by separation on RP-HPLC resulted in the isolation of four active peaks, H1, H2, H3 and H5, inhibiting COX-1, COX-2 and 5-LOX, out of which H3 was identified as 6- hydroxy salvinolone (6-HS). Present findings reveal that PEE of roots of Premna integrifolia exhibits potent anti-inflammatory and immunomodulatory activities, which could form a potential source for development of anti-inflammatory drugs. 6-HS, a COX-2/5-LOX dual inhibitor along with other lead molecules isolated from PEE of Premna integrifolia may form lead molecules for the development of COX-LOX dual inhibitors.

Biochemical properties of a bacterially-expressed Bowman-Birk inhibitor from Rhynchosia sublobata (Schumach.) Meikle seeds and its activity against gut proteases of Achaea janata.

Crude proteinase inhibitors (CPIs) extracted from the seeds of Rhynchosia sublobata, a wild relative of pigeon pea showed pronounced inhibitory activity on the larval gut trypsin-like proteases of lepidopteran insect pest - Achaea janata. Consequently, a full-length cDNA of Bowman-Birk inhibitor gene (RsBBI1) was cloned from the immature seeds of R. sublobata. It contained an ORF of 360 bp encoding a 119-amino acid polypeptide (13.3 kDa) chain with an N-terminus signal sequence comprising of 22 amino acids. The amino acid sequence and phylogenetic analysis together revealed that RsBBI1 exhibited a close relation with BBIs from soybean and Phaseolus spp. A cDNA sequence corresponding to RsBBI1 mature protein (89 amino acid stretch) was expressed in E. coli. The recombinant rRsBBI1 protein with a molecular mass of 9.97 kDa was purified using trypsin affinity chromatography. The purified rRsBBI1 exhibited non-competitive mode of inhibition of both bovine trypsin (Ki of 358 ±â€¯11 nM) and chymotrypsin (Ki of 446 ±â€¯9 nM). Its inhibitory activity against these proteases was stable at high temperatures (>95 °C) and a wide pH range but sensitive to reduction with dithiothreitol (DTT), indicating the importance of disulphide bridges in exhibiting its activity. Also, rRsBBI1 showed significant inhibitory activity (IC50 = 70 ng) on A. janata larval gut trypsin-like proteases (AjGPs). Conversely, it showed <1% inhibitory activity (IC50 = 8 µg) on H. armigera larval gut trypsin-like proteases (HaGPs) than it has against AjGPs. Besides, in vivo feeding experiments clearly indicated the deleterious effects of rRsBBI1 on larval growth and development in A. janata which suggests it can be further exploited for such properties.

Diabetic retinopathy screening programme utilising non-mydriatic fundus imaging in slum populations of New Delhi, India.

OBJECTIVES: To develop and implement a community-based programme for screening of diabetic retinopathy (DR) in urban populations of Delhi. METHODS: Known diabetics (KDs) aged 40 years and older were identified through house-to-house surveys, volunteers and publicity. All KDs were referred to DR screening camps organised locally where procedures included brief medical history, ocular examination and non-mydriatic fundus photography using portable handheld camera. Fundal images were graded on the spot by trained optometrists for DR. Patients with DR were referred to tertiary centre for management. RESULTS: A total of 11 566 KDs were identified, of whom 9435 (81.6%) visited DR screening camps and 8432 (89.4%) had DR gradable images. DR was identified in 13.5% of subjects; 351 cases were mild NPDR, 567 moderate, 92 severe. Seventy-seven had PDR, and 49 had DME, and 2.7% of participants were blind (presenting visual acuity <3/60 in better eye). Non-use of lifestyle management, presence of systemic complications, BMI <18.5 kg/m2 , disease duration of >5 years and uncontrolled diabetes were associated with increased odds of DR. All cases with DR were referred, and 420 (37%) successful referrals to base hospital were observed. CONCLUSION: The programme of creating awareness about DR, identifying KDs and optometrist-led DR screening using non-mydriatic fundus camera based in slums was successful.

Larval Mid-Gut Responses to Sub-Lethal Dose of Cry Toxin in Lepidopteran Pest Achaea janata.

The lack of homogeneity in field application of Bacillus thuringiensis formulation often results in ingestion of sub-lethal doses of the biopesticide by a fraction of pest population and there by promotes the toxin tolerance and resistance in long term. Gut regeneration seems to be one of the possible mechanism by which this is accomplished. However, the existing information is primarily derived from in vitro studies using mid-gut cell cultures. Present study illustrates cellular and molecular changes in mid-gut epithelium of a Bt-susceptible polyphagous insect pest castor semilooper, Achaea janata in response to a Cry toxin formulation. The present report showed that prolonged exposure to sub-lethal doses of Cry toxin formulation has deleterious effect on larval growth and development. Histological analysis of mid-gut tissue exhibits epithelial cell degeneration, which is due to necrotic form of cell death followed by regeneration through enhanced proliferation of mid-gut stem cells. Cell death is demonstrated by confocal microscopy, flow-cytometry, and DNA fragmentation analysis. Cell proliferation in control vs. toxin-exposed larvae is evaluated by bromodeoxyuridine (BrdU) labeling and toluidine blue staining. Intriguingly, in situ mRNA analysis detected the presence of arylphorin transcripts in larval mid-gut epithelial cells. Quantitative PCR analysis further demonstrates altered expression of arylphorin gene in toxin-exposed larvae when compared with the control. The coincidence of enhanced mid-gut cell proliferation coincides with the elevated arylphorin expression upon Cry intoxication suggests that it might play a role in the regeneration of mid-gut epithelial cells.

Insecticidal Effects of Hemocoelic Delivery of Bacillus thuringiensis Cry Toxins in Achaea janata Larvae.

Insecticidal effects of Bacillus thuringiensis Cry toxins in hemocoel of larvae have not been properly evaluated. In the present study, hemocoelic injection of four representative Cry toxins i.e., Cry1Aa, Cry1Ab, Cry1Ac, and DOR5 to an economically important lepidopteran insect pest Achaea janata, induced larval mortality, reduced larval growth rate and gave rise to smaller pupae, all in a dose-dependent manner. We observed extensive degeneration as well as the disintegration of larval tissues, most notably, fat body, and the possible involvement of lysosomal enzymes in tissue histolysis. The resultant "hypoproteinemia" and most relevantly, the drastic reduction of 80-85 kDa hexamerin proteins levels of hemolymph could be attributed to the pathological state of the fat body induced by Cry toxin injection. Formation of non-viable larval-pupal intermediates and emergence of defective adults also indicate toxicity effects of Cry toxins during metamorphosis. Thus, findings from our study suggest Cry toxins in larval hemocoel are also toxic to A. janata larval survival and subsequent development.