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.

MCP-1 binds to oxidized LDL and is carried by lipoprotein(a) in human plasma.

Lipoprotein oxidation plays an important role in pathogenesis of atherosclerosis. Oxidized low density lipoprotein (OxLDL) induces profound inflammatory responses in vascular cells, such as production of monocyte chemoattractant protein-1 (MCP-1) [chemokine (C-C motif) ligand 2], a key chemokine in the initiation and progression of vascular inflammation. Here we demonstrate that OxLDL also binds MCP-1 and that the OxLDL-bound MCP-1 retains its ability to recruit monocytes. A human MCP-1 mutant in which basic amino acids Arg-18 and Lys-19 were replaced with Ala did not bind to OxLDL. The MCP-1 binding to OxLDL was inhibited by the monoclonal antibody E06, which binds oxidized phospholipids (OxPLs) in OxLDL. Because OxPLs are carried by lipoprotein(a) [Lp(a)] in human plasma, we tested to determine whether Lp(a) binds MCP-1. Recombinant wild-type but not mutant MCP-1 added to human plasma bound to Lp(a), and its binding was inhibited by E06. Lp(a) captured from human plasma contained MCP-1 and the Lp(a)-associated endogenous MCP-1 induced monocyte migration. These results demonstrate that OxLDL and Lp(a) bind MCP-1 in vitro and in vivo and that OxPLs are major determinants of the MCP-1 binding. The association of MCP-1 with OxLDL and Lp(a) may play a role in modulating monocyte trafficking during atherogenesis.

The use of biomarkers in the patient with heart failure.

Heart failure is a major burden to the health care system in terms of not only cost, but also morbidity and mortality. Appropriate use of biomarkers is critically important to allow rapid identification and optimal risk stratification and management of patients with both acute and chronic heart failure. This review will discuss the biomarkers that have the most diagnostic, prognostic, and therapeutic value in patients with heart failure. We will discuss established biomarkers such as natriuretic peptides as well as emerging biomarkers reflective of myocyte stress, myocyte injury, extracellular matrix injury, and both neurohormonal and cardio-renal physiology.

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.

Relationship between catheter forces, lesion characteristics, "popping," and char formation: experience with robotic navigation system.

INTRODUCTION: Popping, char and perforation are complications that can occur following catheter ablation. We measured the amount of grams (g) applied to the endocardium during ablation using a sensor incorporated in the long sheath of a robotic system. We evaluated the relationship between lesion formation, pressure, and the development of complications. METHODS: Using a robotic navigation system, lesions were placed in the left atrium (LA) at six settings, using a constant duration (40 seconds) and flow rate of either 17 cc/min or 30 cc/min with an open irrigated catheter (OIC). Evidence of complications was noted and lesion location recorded for later analysis at necropsy. RESULTS: Lesions using 30 Watts (W) were more likely to be transmural at higher (>40 g) than lower (<30 g) pressures (75% vs 25%, P < 0.001). Significantly higher number of lesions using >40 g of pressure demonstrated "popping" and crater formation as compared with lesions with 20-30 g of pressure (41% vs 15%, P = 0.008). A majority of lesions placed using higher power (45 W) with higher pressures (>40 g) were associated with char and crater formation (66.7%). No lesions using 10 g of pressure were transmural, regardless of the power. Lesions placed with a power setting less than 35 W were more likely to result in "relative" sparing of the endocardial surface than lesions at a power setting higher than 35 W (62% vs 33.3%, P = 0.02) regardless of the pressure. CONCLUSIONS: When using an OIC, lower power settings (

Observation of stethoscope sanitation practices in an emergency department setting.

BACKGROUND: Stethoscopes harbor pathogens that can be transferred to patients when proper sanitary measures are not taken. Our aim was to assess medical provider stethoscope cleaning and hand hygiene in an emergency department setting. METHODS: The frequency and methods of stethoscope cleaning during and after provider-patient encounters were observed anonymously in an emergency department of the VA San Diego Healthcare System. RESULTS: Among the total of 426 encounters, 115 (26.9%) involved the use of a personal stethoscope. In 15 of these 115 encounters (13.0%), the provider placed a glove over the stethoscope before patient contact. In 13 of these 115 encounters (11.3%), the provider cleaned the stethoscope with an alcohol swab after patient interaction. Stethoscope hygiene with water and a hand towel before patient interaction was observed in 5 of these 115 encounters (4.3%). Hand sanitizer use or handwashing was observed in 213 of the 426 encounters (50.0%) before patient interaction. Gloves were used before patient interaction in 206 of these 426 encounters (48.4%). Hand sanitizer or handwashing was used in 332 of the 426 encounters (77.9%) after patient interaction. CONCLUSIONS: Rates of stethoscope and hand hygiene performance were lower than expected. Further investigation of stethoscope contamination and the associated risk of nosocomial infection are needed. Perhaps clearer guidelines on proper stethoscope cleaning would reduce this risk.

Esophageal luminal temperature measurement underestimates esophageal tissue temperature during radiofrequency ablation within the canine left atrium: comparison between 8 mm tip and open irrigation catheters.

INTRODUCTION: Evaluation of luminal temperature during left atrial ablation is used clinically; however, luminal temperature does not necessarily reflect temperature within the esophageal wall and poses a risk of atrioesophageal fistula. This animal study evaluates luminal esophageal temperature and its relation to the temperature of the external esophageal tissue during left atrial lesions using the 8 mm solid tip and the open irrigated tip catheters (OIC). METHODS AND RESULTS: A thermocouple was secured to the external surface of the esophagus at the level of the left atrium of the dogs. Luminal esophageal temperature was measured using a standard temperature probe. In four randomized dogs, lesions were placed using an 8 mm solid tip ablation catheter. In six randomized dogs, lesions were placed using the 3.5 mm OIC. The average peak esophageal tissue temperature when using the OIC was significantly higher than that of the 8 mm tip catheter (88.6 degrees C +/- 15.0 degrees C vs. 62.3 degrees C +/- 12.5 degrees C, P < 0.05). Both OIC and 8 mm tip catheter had significantly higher peak tissue temperatures than luminal temperatures (OIC: 88.6 degrees C +/- 15.0 degrees C vs 39.7 degrees C +/- 0.82 degrees C, P < 0.05) (8 mm: 62.3 degrees C +/- 12.5 degrees C vs 39.0 +/- 0.5 degrees C, P < 0.05). Both catheters achieved peak temperatures faster in the tissue as compared to the lumen of the esophagus, although the tissue temperature peaked significantly faster for the OIC (OIC: 25 seconds vs 90 seconds, P < 0.05) (8 mm: 63 seconds vs 105 seconds, P < 0.05). CONCLUSION: Despite the significant difference in actual tissue temperatures, no significant difference was observed in luminal temperatures between the OIC and 8 mm tip catheter.

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.

Internal Transcribed Spacer 1 (ITS1) based sequence typing reveals phylogenetically distinct Ascaris population.

Taxonomic differentiation among morphologically identical Ascaris species is a debatable scientific issue in the context of Ascariasis epidemiology. To explain the disease epidemiology and also the taxonomic position of different Ascaris species, genome information of infecting strains from endemic areas throughout the world is certainly crucial. Ascaris population from human has been genetically characterized based on the widely used genetic marker, internal transcribed spacer1 (ITS1). Along with previously reported and prevalent genotype G1, 8 new sequence variants of ITS1 have been identified. Genotype G1 was significantly present among female patients aged between 10 to 15 years. Intragenic linkage disequilibrium (LD) analysis at target locus within our study population has identified an incomplete LD value with potential recombination events. A separate cluster of Indian isolates with high bootstrap value indicate their distinct phylogenetic position in comparison to the global Ascaris population. Genetic shuffling through recombination could be a possible reason for high population diversity and frequent emergence of new sequence variants, identified in present and other previous studies. This study explores the genetic organization of Indian Ascaris population for the first time which certainly includes some fundamental information on the molecular epidemiology of Ascariasis.

Association between Giardia duodenalis and coinfection with other diarrhea-causing pathogens in India.

Giardia duodenalis, is often seen as an opportunistic pathogen and one of the major food and waterborne parasites. Some insights of Giardia infestation in a diarrhoea-prone population were investigated in the present study. Our primary goal was to understand the interaction of this parasite with other pathogens during infection and to determine some important factors regulating the diarrhoeal disease spectrum of a population. Giardia showed a steady rate of occurrence throughout the entire study period with a nonsignificant association with rainfall (P > 0.05). Interestingly coinfecting pathogens like Vibrio cholerae and rotavirus played a significant (P ≤ 0.001) role in the occurrence of this parasite. Moreover, the age distribution of the diarrhoeal cases was very much dependent on the coinfection rate of Giardia infection. As per our findings, Giardia infection rate seems to play a vital role in regulation of the whole diarrhoeal disease spectrum in this endemic region.

Multilocus sequence typing system (MLST) reveals a significant association of Entamoeba histolytica genetic patterns with disease outcome.

The relationship between parasite genotypes and outcome of amoebic infection is still a paradox and needed to be explored. Proper identification and genetic characterization of Entamoeba histolytica clinical isolates is an effective tool for exploring this relation. Along with conventional polymorphic marker (Chitinase), tRNA linked short tandem repeat (STR) loci has been employed as multilocus genotyping tool due to its better resolution and evolutionary significance. Some common as well as exclusive repeat patterns showing significant relation with disease outcome have been identified. Phylogenetic analysis revealed that repeat patterns exclusively found in asymptomatic and amoebic liver abscess derived isolates are placed in a common lineage and has similar association pattern with the disease outcome. Assumption can be made that isolates of E. histolytica remaining asymptomatic is genetically closer siblings of those causing liver abscess rather than the diarrheal isolates.

The appropriate use of biomarkers in heart failure.

Natriuretic peptides and troponins have been extensively studied and used in heart failure, and their value has been extensively discussed. Renal markers, such as cystasin-C, NGAL, and KIM-1, have shown growing utility in heart failure. The activation of compensatory pathways and ongoing hemodynamic changes result in the release of biomarkers that can be monitored to chart disease progression and possibly target for therapy.

Ablation of atrial fibrillation: what can we tell our patients?

Although catheter-based radiofrequency ablation is no longer experimental, it is not yet the first-line treatment for most patients. The authors describe how this procedure works, its indications, benefits, and limitations, and important points to communicate to potential candidates for this procedure.

Hospital-based surveillance of enteric parasites in Kolkata.

BACKGROUND: Diarrhoea is the second leading cause of illness and death in developing countries and the second commonest cause of death due to infectious diseases among children under five in such countries. Parasites, as well as bacterial and viral pathogens, are important causes of diarrhoea. However, parasitic infections are sometimes overlooked, leading after a period of time to an uncertain aetiology. In this paper we report the prevalence of Giardia lamblia, Entamoeba histolytica and Cryptosporidium sp. in and around Kolkata. FINDINGS: A hospital-based laboratory surveillance study was conducted among the patients admitted between November 2007 and October 2008 to the Infectious Diseases (ID) Hospital (Population = 1103) with diarrhoeal complaints. Of the 1103 samples collected, 147 were positive for Giardia lamblia, 84 for Cryptosporidium sp. and 51 for Entamoeba histolytica. For all these parasites there was a high rate of mixed infection with common enteric viruses and bacteria such as Rotavirus, Vibrio cholerae and Shigella sp. There were also cases of co-infection with all other diarrheogenic pathogens. The age group >or= 5 years had the highest prevalence of parasites whereas the age group >5 - 10 years was predominantly infected with Giardia lamblia (p =< 0.001; Odds ratio (OR) = 3.937; 95% Confidence interval (CI) = 1.862 - 8.326) and with all parasites (p = 0.040; OR = 2.043; 95% CI = 1.033 - 4.039). The age group >10 - 20 years could also be considered at risk for G. lamblia (p = 0.009; OR = 2.231; 95% CI = 1.223 - 4.067). Month-wise occurrence data showed an endemic presence of G. lamblia whereas Cryptosporidium sp. and E. histolytica occurred sporadically. The GIS study revealed that parasites were more prevalent in areas such as Tangra, Tiljala and Rajarhat, which are mainly slum areas. Because most of the population surveyed was in the lower income group, consumption of contaminated water and food could be the major underlying cause of parasitic infestations. CONCLUSION: This study provides important information on the occurrence and distribution of three important intestinal parasites and indicates their diarrheogenic capacity in Kolkata and surrounding areas.