IL10 variant g.5311A is associated with Visceral Leishmaniasis in Indian population

Background: Visceral Leishmaniasis (VL) is a multifactorial disease, where the host genetics play a significant role in determining the disease outcome. The immunological role of anti-inflammatory cytokine, Interleukin 10 (IL10), has been well-documented in parasite infections and considered as a key regulatory cytokine for VL. Although VL patients in India display high level of IL10 in blood serum, no genetic study has been conducted to assess the VL susceptibility/resistance. Therefore, the aim of this study is to investigate the role of IL10 variations in Indian VL; and to estimate the distribution of disease associated allele in diverse Indian populations. Methodology: All the exons and exon-intron boundaries of IL10 were sequenced in 184 VL patients along with 172 ethnically matched controls from VL endemic region of India. Result and Discussion: Our analysis revealed four variations; rs1518111 (2195 A>G, intron), rs1554286 (2607 C>T, intron), rs3024496 (4976 T>C, 3’ UTR) and rs3024498 (5311 A>G, 3’ UTR). Of these, a variant g.5311A is significantly associated with VL (χ2 = 18.87; p = 0.00001). In silico approaches have shown that a putative micro RNA binding site (miR-4321) is lost in rs3024498 mRNA. Further, analysis of the above four variations in 1138 individuals from 34 ethnic populations, representing different social and linguistic groups who are inhabited in different geographical regions of India, showed variable frequency. Interestingly, we have found, majority of the tribal populations have low frequency of VL (‘A’ of rs3024498); and high frequency of leprosy (‘T’ of rs1554286), and Behcet’s (‘A’ of rs1518111) associated alleles, whereas these were vice versa in castes. Our findings suggest that majority of tribal populations of India carry the protected/less severe allele against VL, while risk/more severe allele for leprosy and Behcet’s disease. This study has potential implications in counseling and management of VL and other infectious diseases

Priming exposures to lipopolysaccharides do not affect the induction of Polycomb target genes upon re-exposure

The Polycomb group (PcG) proteins are chromatin factors underlying the process of transcriptional memory to preserve developmental decisions and keep cellular identities. However, not only developmental signals need to be memorized and thus maintained during the life of an organism. For host protection against pathogens, also a memory of previous exposures to an immunogenic stimulus is crucial to mount a more protective immune response upon re-exposure. The antigen-specific adaptive immunity in vertebrates is an example of such a memory to previous immunogenic stimulation. Recently, adaptive characteristics were also attributed to innate immunity, which was classically seen to lack memory. However, the mechanistic details of an adaptive innate immune response are yet to be fully understood and chromatin-based epigenetic mechanisms seem to play an important role in this phenomenon. Possibly, PcG proteins can contribute to such an epigenetic innate immune memory. In this study, we analyzed whether the PcG system can mediate a transcriptional memory of exposure to lipopolysaccharides (LPS). To this end, various forms of LPS pre-treatment were applied to reporter cells and expression kinetics of PcG target genes were analyzed after a second LPS exposure. Neither single nor multiple LPS pre-treatment affected the induction of endogenous LPS-responsive transcripts upon re-exposure. Altogether, our extensive analyses did not provide any evidence for a PcG system-mediated memory of LPS stimulation.ISSN:1932-620

Open-source personal pipetting robots with live-cell incubation and microscopy compatibility

Liquid handling robots have the potential to automate many procedures in life sciences. However, they are not in widespread use in academic settings, where funding, space and maintenance specialists are usually limiting. In addition, current robots require lengthy programming by specialists and are incompatible with most academic laboratories with constantly changing small-scale projects. Here, we present the Pipetting Helper Imaging Lid (PHIL), an inexpensive, small, open-source personal liquid handling robot. It is designed for inexperienced users, with self-production from cheap commercial and 3D-printable components and custom control software. PHIL successfully automates pipetting (incl. aspiration) for e.g. tissue immunostainings and stimulations of live stem and progenitor cells during time-lapse microscopy using 3D printed peristaltic pumps. PHIL is cheap enough to put a personal pipetting robot within the reach of most labs and enables users without programming skills to easily automate a large range of experiments.ISSN:2041-172

Open-source personal pipetting robots with live-cell incubation and microscopy compatibility

Liquid handling robots have the potential to automate many procedures in life sciences. However, they are not in widespread use in academic settings, where funding, space and maintenance specialists are usually limiting. In addition, current robots require lengthy programming by specialists and are incompatible with most academic laboratories with constantly changing small-scale projects. Here, we present the Pipetting Helper Imaging Lid (PHIL), an inexpensive, small, open-source personal liquid handling robot. It is designed for inexperienced users, with self-production from cheap commercial and 3D-printable components and custom control software. PHIL successfully automated pipetting for e.g. tissue immunostainings and stimulations of live stem and progenitor cells during time-lapse microscopy. PHIL is cheap enough for any laboratory member to have their own personal pipetting robot(s), and enables users without programming skills to easily automate a large range of experiments

A. Distribution of genotype frequencies for<i>IL10</i> polymorphism, rs1518111 A>G; rs1554286 C>T; rs3024496 C>T and rs3024498 A>G in VL case and control subjects

<p>. B. Distribution of allele frequencies for <i>IL10</i> polymorphism, rs1518111 A>G; rs1554286 C>T; rs3024496 C>T and rs3024498 A>G in VL case and control subjects.</p

Geographical location and allele distribution of rs1518111 (2195 A>G, intron), rs1554286 (2607 C>T, intron) and rs3024498 (5311 A>G) polymorphism in tribe and caste population of India.

<p>Population no. 1 to 16 belongs to castes (Fig A, C and E). Population no. 17 to 34 belongs to tribes (Fig B, D and F).</p

The structure of the human <i>IL10</i> (chr1, 206945839–206940947; ENST00000423557).

<p>Exons of the gene are shown in pink, introns in brown. rs1518111 (2195 A>G) and rs1554286 (2607 C>T) were the intronic variant of second and third exons while rs3024496 (4976 T>C) and rs3024498 (5311 A>G) were the 3’ UTR variant of fifth exons.</p

Sample size, mean age and sex ratio of case and control samples.

<p>Sample size, mean age and sex ratio of case and control samples.</p

Prevalence (yearly in millions) of different infectious disease (Visceral Leishmaniasis, Leprosy, Tuberculosis Malaria and Filaria) in worldwide and Indian region.

<p>Prevalence (yearly in millions) of different infectious disease (Visceral Leishmaniasis, Leprosy, Tuberculosis Malaria and Filaria) in worldwide and Indian region.</p

Primer sequence, GC % and annealing temperature of <i>IL10</i> exons.

<p>Primer sequence, GC % and annealing temperature of <i>IL10</i> exons.</p