Functional characterization of phospholipase B enzyme from Giardia lamblia.

The microaerotolarent amitochondriate protozoan Giardia lamblia causes Giardiasis and produces a unique enzyme called Phospholipase B (PLB) in contrast to higher eukaryotes. The enzyme is produced upon induction with oxidative (H2O2) stress, thus leading to prostaglandin E2 (PGE2) production. It exists in dimeric form, and its molecular weight is 56 kDa. This PLB was extracellularly cloned in the pET21d vector. The ORF is 1620 bp (Genbank accession no. -OM939681) long and codes for a protein 539 amino acid long, with a 15 amino acid long amino-terminal signal peptide. The highest enzyme activity of PLB was identified at pH 7.5 and 35 °C. This specific enzyme was also active at 50 °C pH 10, but activity was low. We also analyzed the expression of PLB protein in G. lamblia, which was significantly induced under increased oxidative stress.

Domain Adapted Multitask Learning for Segmenting Amoeboid Cells in Microscopy.

The method proposed in this paper is a robust combination of multi-task learning and unsupervised domain adaptation for segmenting amoeboid cells in microscopy. A highlight of this work is the manner in which the model's hyperparameters are estimated. The detriments of ad-hoc parameter estimation are well known, but this issue remains largely unaddressed in the context of CNN-based segmentation. Using a novel min-max formulation of the segmentation cost function our proposed method analytically estimates the model's hyperparameters, while simultaneously learning the CNN weights during training. This end-to-end framework provides a consolidated mechanism to harness the potential of multi-task learning to isolate and segment clustered cells from low contrast brightfield images, and it simultaneously leverages deep domain adaptation to segment fluorescent cells without explicit pixel-level re- annotation of the data. Experimental validations on multi-cellular images strongly suggest the effectiveness of the proposed technique, and our quantitative results show at least 15% and 10% improvement in cell segmentation on brightfield and fluorescence images respectively compared to contemporary supervised segmentation methods.

Epidemiology of Soil-Transmitted Helminth Infections among Primary School Children in the States of Chhattisgarh, Telangana, and Tripura, India, 2015-2016.

Soil-transmitted helminth (STH) infections are highly prevalent in many developing countries, affecting the poorest and most deprived communities. We conducted school-based surveys among children studying in first to fifth standard in government schools in the Indian States of Chhattisgarh, Telangana, and Tripura to estimate the prevalence and intensity of STH infections during November 2015 and January 2016. We adopted a two-stage cluster sampling design, with a random selection of districts within each agro-climatic zone in the first stage. In the second stage, government primary schools were selected by probability proportional to size method from the selected districts. We collected information about demographic details, water, sanitation, and hygiene (WASH) characteristics and stool samples from the school children. Stool samples were tested using Kato-Katz method. Stool samples from 3,313 school children (Chhattisgarh: 1,442, Telangana: 1,443, and Tripura: 428) were examined. The overall prevalence of any STH infection was 80.2% (95% confidence interval [CI]: 73.3-85.7) in Chhattisgarh, 60.7% (95% CI: 53.8-67.2) in Telangana, and 59.8% (95% CI: 49.0-69.7) in Tripura. Ascaris lumbricoides was the most prevalent STH infection in all three states. Most of the STH infections were of light intensity. Our study findings indicate that STH infections were highly prevalent among the school children in Chhattisgarh, Telangana, and Tripura, indicating the need for strengthening STH control program in these states. The prevalence estimates from the survey would serve as a baseline for documenting the impact of the National Deworming Day programs in these states.

3D cell morphology detection by association for embryo heart morphogenesis.

Advances in tissue engineering for cardiac regenerative medicine require cellular-level understanding of the mechanism of cardiac muscle growth during embryonic developmental stage. Computational methods to automatize cell segmentation in 3D and deliver accurate, quantitative morphology of cardiomyocytes, are imperative to provide insight into cell behavior underlying cardiac tissue growth. Detecting individual cells from volumetric images of dense tissue, poised with low signal-to-noise ratio and severe intensity in homogeneity, is a challenging task. In this article, we develop a robust segmentation tool capable of extracting cellular morphological parameters from 3D multifluorescence images of murine heart, captured via light-sheet microscopy. The proposed pipeline incorporates a neural network for 2D detection of nuclei and cell membranes. A graph-based global association employs the 2D nuclei detections to reconstruct 3D nuclei. A novel optimization embedding the network flow algorithm in an alternating direction method of multipliers is proposed to solve the global object association problem. The associated 3D nuclei serve as the initialization of an active mesh model to obtain the 3D segmentation of individual myocardial cells. The efficiency of our method over the state-of-the-art methods is observed via various qualitative and quantitative evaluation.

Pyruvate Protects Giardia Trophozoites from Cysteine-Ascorbate Deprived Medium Induced Cytotoxicity.

Giardia lamblia, an anaerobic, amitochondriate protozoan parasite causes parasitic infection giardiasis in children and young adults. It produces pyruvate, a major metabolic product for its fermentative metabolism. The current study was undertaken to explore the effects of pyruvate as a physiological antioxidant during oxidative stress in Giardia by cysteine-ascorbate deprivation and further investigation upon the hypothesis that oxidative stress due to metabolism was the reason behind the cytotoxicity. We have estimated intracellular reactive oxygen species generation due to cysteine-ascorbate deprivation in Giardia. In the present study, we have examined the effects of extracellular addition of pyruvate, during oxidative stress generated from cysteine-ascorbate deprivation in culture media on DNA damage in Giardia. The intracellular pyruvate concentrations at several time points were measured in the trophozoites during stress. Trophozoites viability under cysteine-ascorbate deprived (CAD) medium in presence and absence of extracellular pyruvate has also been measured. The exogenous addition of a physiologically relevant concentration of pyruvate to trophozoites suspension was shown to attenuate the rate of ROS generation. We have demonstrated that Giardia protects itself from destructive consequences of ROS by maintaining the intracellular pyruvate concentration. Pyruvate recovers Giardia trophozoites from oxidative stress by decreasing the number of DNA breaks that might favor DNA repair.

SDL: Saliency-Based Dictionary Learning Framework for Image Similarity.

In image classification, obtaining adequate data to learn a robust classifier has often proven to be difficult in several scenarios. Classification of histological tissue images for health care analysis is a notable application in this context due to the necessity of surgery, biopsy or autopsy. To adequately exploit limited training data in classification, we propose a saliency guided dictionary learning method and subsequently an image similarity technique for histo-pathological image classification. Salient object detection from images aids in the identification of discriminative image features. We leverage the saliency values for the local image regions to learn a dictionary and respective sparse codes for an image, such that the more salient features are reconstructed with smaller error. The dictionary learned from an image gives a compact representation of the image itself and is capable of representing images with similar content, with comparable sparse codes. We employ this idea to design a similarity measure between a pair of images, where local image features of one image, are encoded with the dictionary learned from the other and vice versa. To effectively utilize the learned dictionary, we take into account the contribution of each dictionary atom in the sparse codes to generate a global image representation for image comparison. The efficacy of the proposed method was evaluated using three tissue data sets that consist of mammalian kidney, lung and spleen tissue, breast cancer, and colon cancer tissue images. From the experiments, we observe that our methods outperform the state of the art with an increase of 14.2% in the average classification accuracy over all data sets.

High prevalence of soil-transmitted helminth infections among primary school children, Uttar Pradesh, India, 2015.

BACKGROUND: Soil-transmitted helminth (STH) infections often affect the poorest and most deprived communities. In order to generate reliable data for planning a school based deworming program, we conducted a survey among primary school children studying in government schools in the Indian state of Uttar Pradesh. The objectives of our survey were to estimate the prevalence and intensity of STH infections. METHODS: We conducted a cross-sectional survey among children studying in 130 primary schools from 9 agro-climatic zones, during May - August 2015. Information about socio-demographic details, defecation and hand-hygiene practices, and stool samples were collected from the school children. Stool samples were examined using the Kato-Katz method. RESULTS: Stool samples from 6421 school children were examined. The overall weighted prevalence of any STH in the State was 75.6% (95% CI: 71.2-79.5). The prevalence was more than 50% in six of the nine agro-climatic zones. A. lumbricoides was the most prevalent STH (prevalence: 69.6%), followed by hookworm (prevalence: 22.6%) and T. trichura (4.6%). The majority of the STH infections were of low intensity. The practice of open defecation and not washing hands with soap after defecation and residence in kutcha house were significant risk factors of STH infection. CONCLUSIONS: STH prevalence among primary school children in Uttar Pradesh was high. Given the WHO guidelines on deworming frequency according to STH prevalence, Govt of Uttar Pradesh needs to implement a school-based deworming program with bi-annual frequency. The findings of our survey would also help monitor the performance of school based deworming programme.