Preliminary analysis of Plasmodium vivax genotypes isolated in southeastern Turkey

WOS: 000351855600009PubMed ID: 26203991Plasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.Ege University, TurkeyEge University [2008TIP024]This study received financial support from the grant given by Ege University, Turkey (Grant number: 2008TIP024) to Yuksel Guruz. All authors read and approved the manuscript and declare that they have no competing interests

The biomarker features of miR-145-3p determined via meta-analysis validated by qRT-PCR in metastatic cancer cell lines

WOS: 000483423500041PubMed ID: 31195093MicroRNAs (miRNAs) play important roles in the cancer biology such as proliferation, differentiation, and apoptosis. The pivotal roles that miRNA expression plays, make them ideal candidates for detection of cancer progression as well as cancer metastasis. Especially for breast, lung and prostate cancer which are originated from soft tissues and prone to metastasis. Thus, the aim of this study is to evaluate the expression level of miR145-3p which is a shared potential biomarker identified by meta-analysis of breast, prostate and lung cancer data sets. Six different data sets representative of three different cancer types were analyzed. These data sets are pooled together to have a master metamiRNA list while getting rid of the platform differentiations between them. As a result, 24 common differentially expressed miRNAs are determined in which miR-145-3p has the topmost rank. To mimic in vivo cancer microenvironment, hypoxia and serum deprivation were used to induce metastasis in breast (MCF-7, MDA-MB-231, MDA-MB-453), prostate (PC3, LNCaP, DU145), lung (A549, NCIH82,) cancer cell lines and noncancerous cell lines of the coresponding tissues (MCF10A, RWPE-1, MRC-5). miR-145-3p expression levels were determined by qRT-PCR. It has been shown that it is down regulated by the induction of metastasis in cancer cell lines while it is up regulated in normal cell lines to suppress the tumor formation. As a conclusion, as representing the same results in three different cancer cell types, miR-145-3p will be a promising biomarker to follow up its expression to detect cancer metastasis

Construction of a multiepitope vaccine candidate against Fasciola hepatica: an in silico design using various immunogenic excretory/secretory antigens

Background Fasciola hepatica is an important pathogen that causes liver fluke disease in definitive hosts such as livestock animals and humans. Various excretory/secretory products have been used in serological diagnosis and vaccination studies targeting fasciolosis. There are no commercial vaccines against fasciolosis yet. Bioinformatic analysis based on computational methods have lower cost and provide faster output compared to conventional vaccine antigen discovery techniques. The aim of this study was to predict B- and T-cell specific epitopes of four excretory/secretory antigens (Kunitz-type serine protease inhibitor, cathepsin L1, helminth defense molecule, and glutathione S-transferase) of Fasciola hepatica and to construct a multiepitope vaccine candidate against fasciolosis. Methods and Results Initially, nonallergic and the highest antigenic B- and T- cell epitopes were selected and then, physico-chemical parameters, secondary and tertiary structures of designed multiepitope vaccine candidate were predicted. Tertiary structure was refined and validated using online bioinformatic tools. Linear and discontinuous B-cell epitopes and disulfide bonds were determined. Finally, molecular docking analysis for MHC-I and MHC-II receptors was performed. Conclusion This multi-epitope vaccine candidate antigen, with high immunological properties, can be considered as a promising vaccine candidate for animal experiments and wet lab studies

Cryptosporidium spp. during chemotherapy: a cross-sectional study of 94 patients with malignant solid tumor

BACKGROUND: Cryptosporidium spp. is a protozoan parasite that infects many vertebrate animals, including humans. Since Cryptosporidium spp. can cause chronic life-threatening diarrhea and severe malabsorption in immunocompromised patients, we investigated the prevalence of this parasite among patients undergoing chemotherapy for malignant solid tumors. OBJECTIVE: Investigate the prevalence of Cryptosporidium spp. in stool samples. DESIGN: Cross-sectional. SETTING: Tertiary care. PATIENTS AND METHODS: Stool samples were collected from adult patients with malignant solid tumors receiving chemotherapy and diarrhea. Cryptosporidium spp. prevalence was determined using Ziehl-Neelsen staining, ELISA, and real-time PCR targeting of the COWP gene. MAIN OUTCOME MEASURE: The prevalence of Cryptosporidium spp. in patients undergoing chemotherapy for malignant solid tumors. SAMPLE SIZE: 94 RESULTS: The prevalence was 2.1% (2/94), 5.3% (5/94), and 5.3% (5/94) as detected by Ziehl-Neelsen staining, real-time PCR and ELISA, respectively. The prevalence reached 8.5% (8/94) using all results obtained from the three methods. Among eight positive stool samples, four were positive by at least two different methods (Ziehl-Neelsen staining-ELISA or ELISA-real-time PCR) whereas the remaining four were positive by either ELISA or real-time PCR. CONCLUSION: These findings show the risk of cryptosporidiosis in cancer patients and the necessity to use at least two diagnostic methods during the diagnosis of cryptosporidiosis to reach more accurate and trustworthy results. LIMITATIONS: Further studies with a larger sample size are recommended. CONFLICT OF INTEREST: None.Izmir Katip Celebi University Scientific Research Projects Coordination Unit [x 2018-TDU-TIPF-0055]Izmir Katip Celebi University Scientific Research Projects Coordination Unit x 2018-TDU-TIPF-0055 to A.A

Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples

Background Discovery of new Toxoplasma gondii serotyping epitopes is important due to reports showing the influence of genotype on the severity of toxoplasmosis. In Turkey, genotypes belonging to type II, type III and Africa 1 lineages were mainly detected. The present study focused on to find out epitopes with high discriminative capacity to serotype these genotypes using well characterized strains isolated from Turkey. Methods To meet this objective, GRA6 and GRA7 genes were sequenced from strains belonging to the type II, III and Africa 1 lineages, and B cell epitopes inside these sequences were predicted by Bcepred and additional docking analysis was performed with B cell receptor. Based on these analyses, 22 peptides harboring lineage specific epitopes were synthesized. Then, the serotyping potency of these peptides was tested using peptide ELISA and well categorized serum samples collected from stray cats infected with genotypes of the different lineages type II (n:9), III (n:1) and Africa 1 (n:1). As a result of peptide-ELISA, a serotyping schema was constructed with peptides that show high discriminative capacity and this assay was validated by sera collected from humans after an outbreak (n:30) and mother/newborn pair sera (n:3). Later, the validated serotyping schema was used to serotype a larger group of human (n:38) and cat (n:24) sera. Results Among 22 peptides, GRA6II/c, GRA7III/d, and GRA6 Africa 1/b epitopes have shown discriminative capacity. During the validation of peptide-ELISA, the serotype of toxoplasmosis outbreak and mother/newborn cases were detected to be serotype II. Moreover, the analyses in a larger group showed that serotype II was prevalent in humans and stray cats. Conclusions Overall, the results showed that the serotyping schema could be successfully used to serotype T. gondii infections caused by type II, III and Africa 1 genotype.This study was supported by a project given by the Izmir Katip Celebi Scientific Research Projects Coordination Unit (Project number: 2018-ONAPTIPF-0006) to AAG.Izmir Katip Celebi Scientific Research Projects Coordination Unit [2018-ONAPTIPF-0006

Investigation of Folding of Purified Recombinant GRA1 Protein Using Web Based Protein Disorder Servers and Trypsin Digestion

WOS: 000268219500018PubMed ID: 19601915The successful folding of a recombinant protein after expression and purification is essential for structural, biochemical and vaccination studies. Toxoplasma gondii recombinant GRA1 protein is a promising vaccine candidate against toxoplasmosis. In the present study, the folding of recombinant GRA1 protein has been evaluated by web based bioinformatics tools that predict protein folding. Subsequently, trypsin digestion, which is a simple indication of proper protein folding, has been used to determine whether recombinant GRA1 protein is likely to be folded. The results indicate that the recombinant GRA1 protein is predicted to be folded by most of the web based bioinformatics predictors. Moreover, in protease digestion experiments, the recombinant GRA1, which was purified to homogeneity without the use of denaturants, gives rise to a discrete band pattern that is indicative of a folded protein. Together, the results suggest that recombinant GRA1 protein is in a folded conformation, suitable for structural, biochemical and vaccination studies

Analysis of Cytidine Monophospho-N-Acetylneuraminic Acid Hydroxylase (CMAH) Gene Related to Neonatal Isoerythrolysis in Stray Cats of Izmir, Turkey

WOS: 000374312100006Neonatal isoerythrolysis is a life threatening disease in new born cats. It occurs when type A or type AB kittens are born from a type B queen (female cat). A homozygous 18 bp insertion located in cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene has been detected in type B cats, causing production of inactive CMAH enzyme. Currently, molecular methods are being used to determine type B blood in female cats, which can help prevent neonatal isoerythrolysis in kittens. These molecular assays target the presence of 18 bp insertion in CMAH gene. In this study, we aimed to analyze the potential of neonatal isoerythrolysis among stray cats of Izmir, Turkey using PCR detecting the 18 bp insertion in CMAH gene. During the study, we analyzed 793 cats' blood sample for the presence of 18 bp insertion in CMAH gene. Three cats known to have blood types A, B, and AB were used as control in PCR. According to the PCR results, blood type A control cat displayed a 175 bp product indicating a homozygous type A cat while blood type control B cat showed a 193 bp product in CMAH gene (with 18 bp insertion) indicating a homozygous type B cat. Interestingly, blood type AB control cat showed a heterozygous pattern for CMAH gene, in which three different bands (175 bp like that of type A, 193 bp product for type B, and the third unique band with approximately 240 bp size) were detected. Among 793 stray cats of Izmir, 791 were homozygous for CMAH gene with 175 bp band size (99.7%). The remaining two stray cats showed heterozygous band pattern like blood type AB cat (0.12%). Overall, 175 bp band displaying type A cats are prevalent contrary to the two cats that have type AB pattern and non-existence of homozygous type B cats. These results show that the potential of neonatal isoerythrolysis in stray cats of Izmir is minimal. Future studies are required to scrutinize the reason(s) for non-existence of type B cats in Izmir and presence of unique band in blood type AB

Do Toxoplasma gondii apicoplast proteins have antigenic potential? An in silico study

Can, Huseyin/0000-0001-9633-9786; Koseoglu, Ahmet Efe/0000-0002-3505-4397WOS: 000510947400018PubMed: 31810853Toxoplasma gondii, one of the extensively studied Apicomplexan parasites, is prevalent worldwide in animals and humans. Apart from its nuclear genome, T. gondii contains an apicoplast genome in 35 kb length which is originated from a secondary endosymbiotic event. in this study, we aimed to investigate the antigenic potential of apicoplast genome encoded proteins (n:28) of T. gondii using in silica analysis. For this purpose, proteins were primarily predicted to reveal antigenic probability and then, several bioinformatics analyses were applied for all predicted antigenic apicoplast proteins to analyze physico-chemical parameters, subcellular localization and transmembrane domain. Also, further prediction analyses including structural, B cell and MHC-I/II epitope sites as well as post-translational modifications were performed for antigenic proteins that have a signal peptide or a high antigenicity value. of the 28 apicoplast proteins, 19 were predicted as probable antigen. Among antigenic proteins, ribosomal protein S5, L11 and S2 were predicted to have signal peptide whereas ribosomal protein L36 and S17 were predicted to have a significantly high antigenicity value (P < 0.05). in addition, ribosomal protein S5, L11, S2, L36 and S17 were predicted to have a lot of epitopes which have low IC50 and percentile rank value indicating a strong binding among epitopes and MHC-I/II alleles, and post-translational modifications such as N-linked glycosylation, acetylation and phosphorylation. To the best of authors' knowledge this is the first study to show the antigenic potential and other properties of apicoplast-derived proteins of T. gondii