Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus granulosus for Diagnosis of Human Hydatidosis

Background: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test.Methods: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individu­als (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen.Results: Recombinant protein was purified by affinity chromatography using His-Tag column. After purifica­tion, recombinant protein was confirmed by western blot analysis using His Tag monoclonal anti­body or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16. Conclusion: While the produced recombinant AgB16 kDa showed promising results in diagnosing human hy­datidosis, but more investigations should be implemented to reach an accurate gold standard

The elusive MAESTRO gene: Its human reproductive tissue-specific expression pattern

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Gene Cloning of 30 kDa Toxoplasma gondii Tachyzoites Surface An¬tigen (SAG1)

Background: Toxoplasma gondii is an obligate intracellular parasite and its sexual and asexual cycles, respectively take place in the intestinal epithelial cell of definitive host and tissue of intermediate hosts. Congenital toxoplasmosis is more impor¬tant when the mother acquired the infection during pregnancy period for the first time. Serological tests are the only meth¬ods for diagnosis of toxoplasmosis. Among serological tests, ELISA has specific value and availability of parasite spe¬cific anti¬gen increases the specificity of test. This study has designed and performed in the aim of availability to specific anti¬gen of Toxoplasma. Methods: A pair of forward and reverse primers was designed based on published sequence of T. gondii SAG1 gene. PCR reac¬tion was performed and PCR product was cloned in the pQE-30 expression vector. Results: The gene of 30 kDa protein of Toxoplasma tachyzoites was cloned in expression vector successfully. Recombinant plas¬mid was confirmed and is ready to express recombinant protein for further studies. Conclusion: In this research we cloned P30 gene of T. gondii tachyzoites surface protein successfully and is ready to ex¬press the recom¬binant protein

In Vitro Differentiation of First Trimester Human Umbilical Cord Perivascular Cells into Contracting Cardiomyocyte-Like Cells

Myocardial infarction (MI) causes an extensive loss of heart muscle cells and leads to congestive heart disease (CAD), the leading cause of mortality and morbidity worldwide. Mesenchymal stromal cell- (MSC-) based cell therapy is a promising option to replace invasive interventions. However the optimal cell type providing significant cardiac regeneration after MI is yet to be found. The aim of our study was to investigate the cardiomyogenic differentiation potential of first trimester human umbilical cord perivascular cells (FTM HUCPVCs), a novel, young source of immunoprivileged mesenchymal stromal cells. Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity. When applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells within 1 week of coculture, making them the first MSC type with this ability. Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting that in vitro predifferentiation could be a potential strategy to increase their effectiveness in vivo.Peer Reviewe

Spirituality and religiosity of non-directed (altruistic) living kidney donors.

AIMS AND OBJECTIVES:To describe the spirituality and religiosity of 30 non-directed (altruistic) living kidney donors in the USA and explore how they may have affected their motivations to donate and donation process experiences. BACKGROUND:The rise in non-directed donors and their ability to initiate kidney chains offer a novel approach to help alleviate the overextended kidney transplant wait list in the USA. However, little is known about the non-directed donors' motivations, characteristics and experiences. DESIGN:We conducted a qualitative-dominant study and used a grounded theory approach to analyse data. METHODS:Thirty participants completed in-depth interviews between April 2013-April 2015. Three analysts independently read and coded interview transcripts. Grounded theory techniques were used to develop descriptive categories and identify topics related to the non-directed donors donation experience. RESULTS:Sixteen of the 30 non-directed donorss discussed the topic of spirituality and religiosity when describing their donation experiences, regardless of whether they were actively practising a religion at the time of donation. Specifically, three themes were identified within spirituality and religiosity: motivation to donate, support in the process, and justification of their donation decisions postdonation. CONCLUSIONS:Findings from this study are the first to describe how spirituality and religiosity influenced the experiences of U.S. non-directed donorss and may help improve non-directed donors educational resources for future spiritual or religious non-directed donors, and the overall non-directed donors donation experience in efforts to increase the living donor pool. RELEVANCE TO CLINICAL PRACTICE:Spirituality and religiosity are often overlooked yet potentially influential factors in Western medicine, as demonstrated through the experiences of Jehovah's Witnesses and their religious restrictions while undergoing surgery and the beliefs of Christian Scientists against taking medications and receiving medical procedures. Understanding needs of non-directed donors specifically with spirituality and religiosity can better position kidney transplant centres and teams to improve predonation screening of non-directed donor candidates and provide support services during the donation process

The elusive MAESTRO gene: Its human reproductive tissue-specific expression pattern

<div><p>The encoded transcript of the Maestro—<i>Male-specific Transcription in the developing Reproductive Organs</i> (MRO) gene exhibits sexual dimorphic expression during murine gonadal development. The gene has no homology to any known gene and its expression pattern, protein function or structure are still unknown. Previously, studying gene expression in human ovarian cumulus cells, we found increased expression of <i>MRO</i> in lean-type Polycystic Ovarian Syndrome (PCOS) subjects, as compared to controls. In this study, we examined the <i>MRO</i> splice variants and protein expression pattern in various human tissues and cells. We found a differential expression pattern of the <i>MRO</i> 5’-UTR region in luteinized granulosa-cumulus cells and in testicular tissues as compared to non-gonadal tissues. Our study also shows a punctate nuclear expression pattern and disperse cytoplasmic expression pattern of the MRO protein in human granulosa-cumulus cells and in testicular germ cells, which was later validated by western blotting. The tentative and unique features of the protein hampered our efforts to gain more insight about this elusive protein. A better understanding of the tissue-specific <i>MRO</i> isoforms expression patterns and the unique structure of the protein may provide important insights into the function of this gene and possibly to the pathophysiology of PCOS.</p></div

<i>MRO</i> expression in human tissues with FL-248.

<p><i>MRO</i> expression in (A) ovarian tissue of a post-menopausal subject with <b>no active folliculogenesis</b> (age 52); (B) adult normal testis; (C) Typical testicular seminoma; (D) Cerebrum (brain) tissue. Abbreviations: GLC–granulosa-lutein cells, TL–theca lutein cells, SC–Sertoli cells, SG–spermatogonia, PS–primary spermatocyte, SP–spermatids, NP–Neuropil, GL–glial cells. Bar ruler, 100um.</p

Clinical parameters of PCOS and control patients.

<p>Clinical parameters of PCOS and control patients.</p