Transcriptomic and epigenomic effects of insoluble particles on J774 macrophages.

Here we report epigenomic and transcriptomic changes in a prototypical J774 macrophage after engulfing talc or titanium dioxide particles in presence of estrogen. Macrophages are the first immune cells to engage and clear particles of various nature. A novel paradigm is emerging, that exposure to so-called 'inert' particulates that are considered innocuous is not really free of consequences. We hypothesized that especially the insoluble, non-digestible particles that do not release a known hazardous chemical can be underappreciated agents acting to affect the regulation inside macrophages upon phagocytosis. We performed gene chip microarray profiling and found that talc alone, and especially with oestrogen, has induced a substantially more prominent gene expression change than titanium dioxide; the affected genes were involved in pathways of cell proliferation, immune response and regulation, and, unexpectedly, enzymes and proteins of epigenetic regulation. We therefore tested the DNA methylation profiles of these cells via epigenome-wide bisulphite sequencing and found vast epigenetic changes in hundreds of loci, remarkably after a very short exposure to particles; ELISA assay for methylcytosine levels determined the particles induced an overall decrease in DNA methylation. We found a few loci where both the transcriptional changes and epigenetic changes occurred in the pathways involving immune and inflammatory signalling. Some transcriptomic and epigenomic changes were shared between talc and titanium dioxide, however, it is especially interesting that each of the two particles of similar size and insoluble nature has also induced a specific pattern of gene expression and DNA methylation changes which we report here.

Evaluation of Og4C3 antigen ELISA as a tool for detection of bancroftian filariasis under lymphatic filariasis elimination programme.

Diagnosis of Lymphatic Filariasis by microscopic examination of thick blood films (TBF) collected between 8.30 pm to 12 midnight, though highly specific is operationally problematic. We evaluated the TropBio Og4C3 serum ELISA as a tool for detection of W. bancrofti microfilaria carriers using Dried Blood Spots (DBS). The study was carried out in two parts (i) to test the sensitivity and specificity of the ELISA test for detection of circulating filarial antigen (CFA) in microfilaria (Mf) carriers vis-à-vis the conventional thick blood film (TBF) microscopy and its persistence in different categories of individuals during the course of disease viz., Endemic normals (n=51), microfilaria (Mf) carriers (n=27), acute cases (n=27), chronic cases (n=50) and a control group of non-endemic normals (n=48) using sera samples and ii) to study the utility of finger prick Dried Blood Spots (DBS) collected on filter paper for detection of Mf carriers and its comparison with another antigen detection assay, the Immunochromatographic test (ICT). Considering the non-endemic normals and microfilaria carriers, the ELISA test was found to have 100% sensitivity and 94.12% specificity for detection of Mf carriers in sera samples. The CFA was absent in majority of the subjects tested under other categories with a positivity of 7.8% among endemic normals, 11.12% among acute cases, 7.84% among chronic cases and 6.25% among nonendemic normals. Comparison of finger prick DBS and sera samples by ELISA vis-à-vis the ICT, carried out on Mf carriers (n=91) and endemic normals (n=97), showed a positivity of 88 (96.7%) in DBS as against 86 (94.5%) in sera samples and 88 (96.7%) by ICT, amongst Mf carriers, with a statistically significant correlation in antigen units between sera and DBS samples (r = 0.959, p = 0.000) amongst the microfilaria carriers. Out of 97 endemic normals, 19 (19.6%) sera and 17 (17.5%) DBS samples tested positive by ELISA while as 12(12.4%) tested positive by ICT, again with a statistically significant correlation between the antigen units in sera and DBS samples (r = 0.942, p = 0.000). DBS prepared from 25 microl of blood were found to be as sensitive as 50 microl for antigen detection. Antigen positivity detected from DBS collected during day and night from known microfilaria carriers (n=27) showed a statistically insignificant difference (p = 0.125) and a significant correlation in antigen units (r = 0.820 and p = 0.013). In view of the comparable results of ELISA, ICT and TBF microscopy, it is concluded that the TropBio Og4C3 ELISA using finger prick DBS can be used as an alternate to TBF microscopy for detection of bancroftian Filariasis under the LFE programme.

Status of lymphatic filariasis in some select slum clusters of Delhi.

Filaria surveys conducted in some select slum clusters namely Hari Nagar, Yamuna pusht near Vijaya Ghat along the Ring Road and Timarpur in Delhi during 1989, 1991 and 1992 respectively, covering a population of approximately 5000 slum dwellers revealed the presence of bancroftian microfilaria (mf) carriers and disease cases. The mf and disease rates (per cent) in these three slum areas were in the order of 6.3, 2.2, 3.7 and 1.4, 0.5 and 0.1 respectively. The mf density varied from 3.1 to 12.3 per 20 cumm. blood. High ten man hour densities of Culex quinquefasciatus (581) in Yamuna pusht followed by (355) in Timarpur were recorded during entomological investigations. Hari Nagar accounted for least ten man hour density of Cx. quinquefasciatus (160), because collection was made during winter months (November-December). The dissection of Cx. quinquefasciatus did not reveal any human filarial infection except in Yamuna pusht where out of 139 only one Cx. quinquefasciatus was found infective.

Persistence of Vibrio cholerae in inter epidemic period--preliminary observations on analysis of water.

Out of the 61 water samples collected from hand pumps and wells from cholera endemic areas of Varanasi City, Vibrio cholerae non 01 was detected in only one sample. However, seven (18.9 per cent) samples out of 37 samples of river water were positive for V. cholerae non 01. None of the samples showed Vibrio cholerae. These observations indicate transmission and dilution of Vibrio cholerae bacillus in environment.

Clinico-epidemiological profiles of post-kala-azar dermal leishmaniasis in Varanasi.

Twenty-seven cases of Post Kala-Azar Dermal Leishmaniasis (P. K. D. L.) were detected in an endemic focus of Kala-azar in Sujabad village in Varanasi Distt. Male-Female ratio of cases was 4.4:1. Majority (66.6 per cent) of cases had macular lesions. Histopathology of one case showed Leishmania donovani (L. D.) bodies. Densities of sand fly were more in pockets where P. K. D. L. cases were detected. All the 13 cases, which were treated with sodium antimony gluconate, responded well to therapy.