Seroprevalence of hepatitis B virus and hepatitis C virus co-infection in human immunodeficiency virus infected patients at a tertiary care hospital in South India

Background: About one third of human immunodeficiency virus (HIV) infected patients are co infected with either hepatitis B virus (HBV) or hepatitis C virus (HCV) as the three viruses have similar routes of transmission that is through transfusion of blood and blood products, sharing of needles to inject drugs and unprotected sexual activity. The survival of HIV infected patients has been markedly improved with highly active antiretroviral therapy (HAART). However several studies showed that the liver diseases caused by HBV or HCV have emerged as one of the leading causes of non AIDS related deaths in HIV patients. The objective of this work was to study the prevalence of HBV & HCV co-infection in HIV infected patients at a Tertiary care centre in South India.Methods: The study group includes 100 HIV seropositive individuals confirmed by three rapid tests as per NACO (National AIDS Control Organization) guidelines in ICTC (Integrated Counseling and Testing Centre), Department of Microbiology, Andhra Medical College, Visakhapatnam, Andhra Pradesh, India. Age and sex matched 100 HIV seronegative individuals were also included in the study as controls. Both the groups were screened for detection of HBV and HCV markers by one rapid test and a solid phase enzyme linked immunosorbent assay (sandwich ELISA).Results: Out of 100 HIV positive patients in the study group 12(12%) were co infected with HBV and 2(2%) were co infected with HCV. Out of 12 HIV and HBV co infected patients 7(58.3%) were females and 5(41.7%) were males. The HIV &HCV co infected patients were both females. Co infection of HBV & HCV with HIV was found to be 0(0%). Co infection was most commonly seen in the age group 31-40 years followed by 21 – 39 years. In the control group out of 100 HIV negative individuals, 1(1%) was infected with HBV infection.Conclusions: The routine screening of HBV and HCV should be mandatory for HIV infected patients, as there is more chance of co infection with these Hepatitis viruses due to enhanced immunodeficiency by HIV and similar routes of transmission. Clear National policies should be established which should include clear economic and health care strategies to improve quality of living conditions, education and easy access to health care facilities.

Seroprevalence of syphilis in human immunodeficiency virus patients

Background: Syphilis is a sexually transmitted infection caused by, Treponema pallidum. Syphilis facilitates the transmission and acquisition of human immunodeficiency virus (HIV) and causes transient increase in the viral load. Sexually transmitted infections (STI) are 3-5 times more likely to acquire HIV infection, if exposed to the virus through sexual contact. Aim of the study was to estimate the seroprevalence of Syphilis in HIV patients.Methods: A total of 920 blood samples were collected from HIV patients attending ART (Antiretroviral therapy) centre and were tested for Syphilis by using Rapid Plasma Reagin (RPR) and Treponema pallidum Hemagglutination Assay (TPHA). A total of 100 HIV non-reactive individuals were taken as a control group.Results: Out of 920 samples, 102 (11.1%) were positive for Syphilis. Out of 102 Syphilis seropositive patients, males (76.5%) were more commonly affected in age group of 21-40 years. Both RPR and TPHA were reactive in 46% of cases and only TPHA reactive in 53.9% of cases. Out of 100 HIV non-reactive patients, 5% of patients are reactive for Syphilis.Conclusions: In the present study, prevalence of Syphilis was more in HIV patients compared to HIV non-reactive persons. Persons with HIV infection acquired through sexual route should be screened for Syphilis by one nonspecific test along with specific test to confirm the diagnosis. This will help in proper management of the patients having Syphilis and HIV co-infection

AN OUTBREAK OF SCRUB TYPHUS IN A REMOTE VILLAGE-AN OVERVIEW

Objective: An outbreak of scrub typhus was declared by Public Health authorities in our district in October, 2022. To investigate this a team of experts from Microbiology, SPM, Medicine and Pediatric Medicine was sent to the place of outbreak and they observed that there was heavy vegetation, stagnant water, dwellings heavily infested with rodent population. There was heavy rain in the past two weeks in the district. To investigate the cause of sudden upsurge of febrile illness cases. Methods: Blood samples were collected from 28 persons who were symptomatic with fever/headache/diarrhea/rash and myalgia to do tests for Scrub typhus, Typhoid fever, Malaria, Dengue, Complete blood count, CRP, LFT and KFT. The samples which were positive for Scrub typhus in rapid test kits were subjected for IgM ELISA to confirm scrub typhus. Results: Seven of twenty-eight patients tested positive for scrub typhus by rapid test (25%), and four of them were positive by IgM ELISA (14.28%). Three patients were Widal-positive (10.7%). More than half samples showed increased CRP levels (53.57%). Thrombocytopenia and mild leucocytosis was observed in scrub typhus cases (42.85%) as well as in typhoid cases. Conclusion: Any outbreak during monsoon should be investigated thoroughly not only for the specified disease but also for all infectious diseases that are prevalent in that area

The <em>C. elegans</em> Rab Family: Identification, Classification and Toolkit Construction

<div><p>Rab monomeric GTPases regulate specific aspects of vesicle transport in eukaryotes including coat recruitment, uncoating, fission, motility, target selection and fusion. Moreover, individual Rab proteins function at specific sites within the cell, for example the ER, golgi and early endosome. Importantly, the localization and function of individual Rab subfamily members are often conserved underscoring the significant contributions that model organisms such as <em>Caenorhabditis elegans</em> can make towards a better understanding of human disease caused by Rab and vesicle trafficking malfunction. With this in mind, a bioinformatics approach was first taken to identify and classify the complete <em>C. elegans</em> Rab family placing individual Rabs into specific subfamilies based on molecular phylogenetics. For genes that were difficult to classify by sequence similarity alone, we did a comparative analysis of intron position among specific subfamilies from yeast to humans. This two-pronged approach allowed the classification of 30 out of 31 <em>C. elegans</em> Rab proteins identified here including <em>Rab31/Rab50</em>, a likely member of the last eukaryotic common ancestor (LECA). Second, a molecular toolset was created to facilitate research on biological processes that involve Rab proteins. Specifically, we used Gateway-compatible <em>C. elegans</em> ORFeome clones as starting material to create 44 full-length, sequence-verified, dominant-negative (DN) and constitutive active (CA) <em>rab</em> open reading frames (ORFs). Development of this toolset provided independent research projects for students enrolled in a research-based molecular techniques course at California State University, East Bay (CSUEB).</p> </div

A chladogram of Rab family members from <i>C. elegans</i> and <i>H. sapien</i>s.

<p>The evolutionary history was inferred using the Neighbor-Joining phylogenetic reconstruction method. The tree is rooted with the natural outlying clade, Rab28. The optimal tree is shown with the percentage of replicate trees (>40) in which the associated genes cluster together in the bootstrap test (500 replicates) provided next to each branch. The tree is drawn to emphasize topology. The evolutionary distances were computed using the JTT amino acid substitution method and are in the units of the number of amino acid differences per site. Evolutionary analyses were conducted using MEGA5. Clades marked with red, orange or yellow circles indicate their degree of stability under a variety of phylogenetic reconstruction parameters (see text and methods for details). Red = 14/14, orange = 13/14 and yellow = 12/14 trees. Genes highlighted with black circles represent putative orphan <i>C. elegan</i> Rabs (lacking a human ortholog). For simplicity, closely related splice variants and well-supported human-specific clades were deleted (see methods for details).</p

Comparative analysis of intron position among diverse Rab subfamily members.

<p>A) Cladogram indicating evolutionary relationships of 18 species examined here <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049387#pone.0049387-Dunn1" target="_blank">[128]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049387#pone.0049387-Csuros1" target="_blank">[130]</a>. Ecdy. = Ecdysozoa, Chromal. = Chromalveolata. For species abbreviations see Methods. B) An ML tree of Opisthokonts created from the MSA used to map intron positions. Bootstrap support (100 replicates) is indicated for each subfamily cluster. C) For each subfamily, the number of times a <u>S</u>ubfamily <u>S</u>pecific <u>C</u>onserved <u>I</u>ntron <u>P</u>osition (SSCIP) involving the indicated number of species was observed (gray bars), compared to what is expected by chance (black diamonds). The difference between observed and expected is statistically significant where indicated. *P(Monte Carlo) <0.05. ***P(Monte Carlo)≤0.00001. The Rab<i>31, 6, 5, 22, 34, 21</i> and <i>23</i> subfamilies include 17, 18, 17, 9, 9, 10, 14 and 12 species, respectively. D) and E) Heat map indicating number of introns within <i>Rab31</i> (D) or <i>Rab6</i> (E) that match SSCIPs from <i>Rab31, 5, 22, 21, 6, 34</i> and <i>23</i>. The circled number indicates the number of introns present in the MSA for each gene. % equals the percentage of introns that are shared with the true SSCIP. <i>C56E6.2</i> (D) and <i>Y71H2AM.12</i> (E) are highlighted red. Genbank Descriptions (if any) and RABDB! classifications are included. Classification abbreviations include: HypoRabX1 (H.RabX1), HypoRabX2 (H.RabX2), HypoRabX3 (H.RabX3) and MetazoaRabX3 (M.RabX3). F) A pairwise comparison of intron position conservation between specific genes (Rab31 at left, Rab6 at right) and their corresponding set of SSCIPs. Black diamonds plot the probability that a specific number of intron matches would be expected by chance for each set of conditions. Chart 1 plots a comparison of 5 introns with 7 SSCIPs (5×7). Chart 2∶4×7. Chart 3∶2×7. Chart 4∶4×6. Observed values for a subset of genes are indicated with P values estimated from the Monte Carlo simulation data (See text and methods). Species abbreviations are as in A. C) and F) 72 protosplice sites assumed.</p

A flow chart describing the lab module involving verification and modification of ORFeome Rab clones.

<p>Steps 1 through 4 were done in parallel to steps A through D. Two peer-review steps at 3 and D were included to minimize mistakes in primer design and sequence analysis of WT ORFeome clones. Abbreviations: Gene of Interest (GOI), Constitutive Active (CA), Dominant Negative (DN), Restriction Fragment Length Polymorphism (RLFP), Polyacrylamide Gel Electrophoresis (PAGE), Human Ras (HRAS), Wild-Type (WT).</p

A list and description of Rab isolates created for the <i>C. elegans</i> ORFeome-based toolkit.

<p>WT, DN and CA clones included in the Rab Toolkit are given isolate names otherwise an explanation for its absence is provided. Subfamily classifications are based on Diekmann et al. 2011 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049387#pone.0049387-Diekmann1" target="_blank">[33]</a> and/or data presented here. The majority of <i>C. elegans rab</i> genes are predicted to have only one splice variant with the following exceptions: WormBase describes two splice variants for <i>rab-3</i> that code for proteins 233 and 219 amino acids (aa) in length. ORFeome project primers were designed to amplify the shorter isoform only. WormBase describes two splice variants for <i>4R79.2 (Rab44)</i> that code for proteins 311 aa and 395 aa in length. ORFeome project primers were designed to amplify the longer isoform only. Names listed under “other” are from WormBase or Pereira-Leal and Seabra (2001). Finally, while <i>rab-37</i> shows 100% identity with the Refseq protein NP_001041293 it contains an additional 5 amino acids at its N-terminus. See text for details.</p