Unusual vascular cause of syncope in an adult patient: type 2 proatlantal intersegmental artery.

A proatlantal intersegmental artery anomaly may present with symptoms such as dizziness and syncope. Because it has the risk of damage in surgical and interventional radiological procedures, to know the presence of this unusual vascular anomaly is very important.

Genetic characterization of Toxoplasma gondii strains isolated from humans living in Izmir, Türkiye.

PURPOSE: Toxoplasma gondii is an obligate intracellular zoonotic parasite that can infect all warm-blooded animals, including humans. Currently, clinical findings of toxoplasmosis are being related to T. gondii strains such as Type I genotype may cause high pathogenicity and Type II genotype causes a milder clinical presentation. We have showed in our previous that Type II genotype is the most frequent strain detected in stray cats and wild birds living in natural life of Izmir. The aim of this study was to assess toxoplasmosis seroprevalence in immunocompromised patients, investigate the presence of T. gondii DNA in their blood samples, and genotype the PCR positive ones. METHODS: The 42 buffy-coat and serum samples were collected from immunocompromised patients who were from various clinics. Thereafter, Real-Time PCR targeting RE gene of T. gondii was performed with DNA samples obtained from buffy-coat samples. Genotyping was performed by sequencing of GRA6 and GRA7 gene regions of positive DNA samples obtained from tissues of bioassay and PCR positive samples. RESULTS: According to Real-Time PCR results, T. gondii DNA was detected in 23.8% (10/42) samples. Among these 10 samples, two samples were determined as T. gondii Type II genotype. Anti-Toxoplasma IgG antibodies were detected in 28.57% (12/42) samples. CONCLUSIONS: Overall, the detection of Type II genotype in humans in Izmir province suggested that T. gondii infection in humans, stray cats, and wild animals may be associated to each other in terms of transmission.

Importance of screening severe COVID-19 patients for IFN-λ1, IL-6 and anti-S1 IgG levels.

Cytokine storm is an important cause of death in COVID-19 patients. A recent clinical study showed that administration of recombinant interferon lambda 1 (IFN-λ1 or IL-29) may prevent severe COVID-19. On the other hand, IL-6 has been associated as a prognostic marker of worsening for COVID-19 patients. The objective of this study is to screen IFN-λ1, IL-6 and antibody levels in consecutive serum sample sets of COVID-19 patients. A total of 365 serum samples collected from 208 hospitalized COVID-19 patients were analyzed for IFN-λ1 and IL-6 levels as well as SARS-CoV-2 neutralizing antibodies and anti-S1 IgG antibodies. Analyses of serum samples for cytokine levels showed that IFN-λ1 (>8 pg/mL) and IL-6 (>2 pg/mL) were detected in approximately 64% and 21% patients, respectively. A decrement in IFN-λ1 levels and IL-6 levels above 35 pg/mL can be sign of clinical severity and upcoming dead. An increment in IL-6 levels wasn't detected in every COVID-19 patient but a decrement in IL-6 levels was related to clinical improvement. Importantly, the detection of IFN-λ1 level together with an increase in anti-S1 IgG antibody response were observed in clinically improved patients. Screening severe COVID-19 patients for IFN-λ1, IL-6, and anti-S1 IgG antibody levels during their hospital stay especially in intensive care units may be beneficial to monitor the clinical status and management of treatment strategies. Importantly, detection of IFN-λ1 together with protective IgG antibody response can be an indication of clinical improvement in severe COVID-19 patients and these patients may be discharged from the hospital soon.

A preliminary study to develop a lateral flow assay using recombinant GRA1 protein for the diagnosis of toxoplasmosis in stray cats.

Toxoplasma gondii is a protozoan parasite that may infect many mammals including humans. Cats are one of the main sources of infection for humans. Therefore, routine screening of cats with tests that are inexpensive, rapid, and do not require sophisticated laboratory equipment is important. In this study, a lateral flow assay (LFA) was designed to rapidly diagnose toxoplasmosis in cats. For this purpose, we selected GRA1 protein of T. gondii due to its high antigenicity in diagnostic and vaccine studies. We further analyzed the immunological properties of GRA1 protein using in silico tools. Then, we expressed and purified recombinant GRA1 (rGRA1) protein and used it during the development of LFA to detect toxoplasmosis in serum samples (n = 40) of cats. According to the results, rGRA1 protein has negative GRAVY value, high aliphatic index, alpha helix, random coil and 12 B cell epitopes. The in silico data supported the high antigenic properties of rGRA1 protein and showed that it can be a good antigen candidate for LFA. Among 30 cat positive serum samples, 27 were found positive by the LFA while seronegative sera (n = 10) were negative by the LFA. The preliminary data showed that the LFA has high sensitivity (90 %) and specificity (100 %). When we used high responsive cat sera (i.e. sera that have optical density > 0.5 with ELISA) the sensitivity value reached 100 %. These results showed that rGRA1 protein is a good candidate to develop a LFA for rapid diagnosis of toxoplasmosis in cats.

Biotechnological Based Recombinant Protein Vaccines Developed Against Toxoplasmosis

Toxoplasma gondii (T. gondii) that can infect most warm-blooded animals and humans, is an obligate intracellular apicomplexan parasite with a wide host range. About one-third of the world's population is infected with this parasite. While toxoplasmosis progresses asymptomatically in individuals with a strong immune system, it can cause serious clinical manifestations and death in immunocompromised individuals. The parasite is transmitted to humans through the consumption of water and food contaminated with cat feces, as well as raw or undercooked animal products, congenital infection and blood/organ transplantation. Additionally, T. gondii is often observed in farm animals such as sheep and goats. Clinical manifestations and abortions caused by T. gondii in sheep and goats lead to enormous economic loss worldwide. There is a commercial vaccine against T. gondii, called Toxovax (MSD, New Zealand) that can only be used in sheep. For these reasons, there is a need for innovative T. gondii vaccine that is harmless, easily produced, which can prevent losses and be used in all living things. Advances in immunology, molecular biology, genetic, biotechnology and proteomics bring new perspectives to vaccine studies. Studies in innovative vaccine studies against T. gondii have accelerated with the discovery of new antigens by in vitro screenings, and bioinformatic analyzes, the use of various expression systems and new adjuvant types. Recombinant protein vaccines are biotechnological vaccines that are frequently preferred due to their rapid and easy production in various expression systems, availability of very and high purity products, ease of manipulation and stimulation of both cellular and humoral immune responses. Recombinant protein vaccines, developed by biotechnological methods, are promising tools for providing a protective immune response against toxoplasmosis. In this review, an overview of the parasite complex life cycle, its pathogenesis, humoral and cellular immune responses in the host, and recombinant protein vaccine studies developed against the parasite are presented.

An Overview of DNA Vaccines Development Studies Against Toxoplasma gondii

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that can infect almost all warm-blooded animals, including humans, and one-third of the global population is thought to be infected with this parasite. Infection can occur through consumption of contaminated food, contact with an infected host, or congenital transmission. While toxoplasmosis is asemptomatic in people with a healthy immune system, it can cause severe infections in people with a suppressed immune system or with immunodeficiency. In addition to causing diseases in humans, it also causes infections in livestock and may result in stillbirth and abortion in sheep and goats. There is no 100% effective medicine or vaccination against the parasite that causes major clinical symptoms and financial losses. There is a need for an effective, safe, and durable vaccine that can provide protective immunity for use in humans and animals. Vaccination studies against toxoplasmosis have gathered speed since the 1990s. Today, studies can be carried out to develop effective and safe vaccines depending on the developments in molecular biology, biotechnology, and immunology. DNA vaccines are a promising vaccine platform against toxoplasmosis because they are easy to produce, they are safe, they do not need a cold chain, and they can stimulate both humoral and cellular immune responses. This review provides an overview of the complex life cycle, pathogenesis, and epidemiology of the parasite; the immune response that develops in the host against the infection it causes; and the DNA vaccines developed against toxoplasmosis and these vaccines.

Development of multistage recombinant protein vaccine formulations against toxoplasmosis using a new chitosan and porin based adjuvant system.

Toxoplasmosis is a global health problem affecting both human and animal populations. The lack of effective treatment makes the development of a vaccine against toxoplasmosis one of the main goals in the management of this disease. In our study, vaccine formulations containing the multistage recombinant antigens, rBAG1 + rGRA1 were developed with a combined adjuvant system consisting of chitosan and Salmonella Typhi porins in micro (MicroAS) and nanoparticulate (NanoAS) forms. BALB/c mice were immunized intraperitoneally with vaccine formulations two times at three-week intervals. Three weeks after the second vaccination, mice were challenged with 7-8 live tissue cysts of the virulent T. gondii PRU strain by oral gavage. Higher cellular uptake by macrophages and enhanced cellular (IFN-γ and I-4 in stimulated spleen cells) and humoral (IgG, IgG1, IgG2a) responses were obtained with the adjuvanted formulation, higher with microsystem when compared to that of nanosystem. Microsystem was found to stimulate Th1-polarized immune responses, whereasnon-adjuvanted antigens stimulated Th2-polarized immune response. The highest survival rate and reduction in cysts numbers and T. gondii DNA were obtained with the adjuvanted antigens.Our study showed that adjuvanted multistage recombinant vaccine systems increase theimmune response with strong protection againstT. gondii, more profoundly in microparticulate form.

Assessment of ADAM17 and ADAM10 proteins with CXCL10 and thyroid autoimmunity in vitiligo pathogenesis.

Introduction: Vitiligo is an acquired chronic pigmentation disorder. The etiopathogenesis is still not fully understood. Aim: To research the correlation of ADAM proteins, shown to be associated with autoimmune diseases like rheumatoid arthritis and lupus erythematosus, with vitiligo also considered to be an autoimmune disease. Material and methods: The study included a patient group of 45 patients with the diagnosis of vitiligo and a control group of 45 healthy adults. The ADAM10 and ADAM17 protein serum levels and CXCL10 and thyroid autoantibody anti-TG and anti-TPO levels along with FT3, FT4, and TSH hormone levels were determined with the ELISA method. Statistical analysis of results was made with the SPSS 22.0 program. Results: In vitiligo patients, the ADAM10 levels (2.34 ±0.80 pg/ml) were statistically significantly low compared to the control group (10.29 ±1.71 pg/ml) (p < 0.05), while the ADAM17 levels (128.51 ±14.37 pg/ml) were statistically significantly high compared to the control group (16.30 ±6.31 pg/ml) (p < 0.05). Additionally, the CXCL10 levels were observed to be statistically significantly higher in the patient group (275.11 ±62.36) than in the control group (107.08 ±33.12). Thyroid autoimmunity test results (anti-TG, anti-TPO, and TSH) were shown to be different to a statistically significant degree in the patient group compared to the control group (p < 0.001, p < 0.000, p = 0.003, respectively). Statistical analyses used the Kolmogorov-Smirnov, Mann-Whitney U test, and the independent T-test. Conclusions: We obtained data that are important in terms of understanding the pathogenesis. ADAM10 and ADAM17 proteins may be new targets for future therapeutic approaches.

The importance of a disintegrin and metalloproteinase (ADAM)10 and ADAM17 in the pathogenesis of psoriasis.

BACKGROUND: Psoriasis is a chronic inflammatory skin disorder characterized by inflammation, hyperproliferation and neoangiogenesis. The disease pathogenesis has not been fully elucidated. The proteins, a disintegrin and metalloproteinase (ADAM)10 and ADAM17, are important proteases serving as regulators of inflammation. AIM: To determine the role of ADAM10 and ADAM17 in the pathogenesis of psoriasis through the comparison of their serum levels in patients with psoriasis and healthy controls (HCs). METHODS: In total, 179 participants (90 patients with psoriasis and 89 HCs) were enrolled in the study. Levels of ADAM10 and ADAM17 in serum were measured by ELISA for each participant from the patient and HC groups. The statistical data analysis was performed using SPSS (V19.0) and P < 0.05 was considered statistically significant. RESULTS: The mean values for serum ADAM10 and ADAM17 were, respectively, 3.1 ± 2.2 and 76.5 ± 31.1 in the psoriasis group and 8.6 ± 3.7 and 29.5 ± 22.4 in the HC group. A statistically significant difference between the patient and HC groups was detected for both ADAM10 and ADAM17 levels (P = 0.001). CONCLUSION: Considering the high levels of ADAM17 in the psoriasis group, ADAM17 protease might have a crucial role in the pathogenesis of psoriasis, while the low levels of ADAM10 might be attributable to its regulatory effect on keratinocyte differentiation and proliferation.

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.

Rapid detection of Toxoplasma gondii DNA in cat feces using colorimetric loop-mediated isothermal amplification (LAMP) assays targeting RE and B1 genes.

Toxoplasma gondii is an obligate protozoan parasite that can infect mammals and birds. Cats are the definitive host of T. gondii and have a very important role in transmission of toxoplasmosis due to the shedding of millions of unsporulated oocysts, that become infective in the environment. Since cats play a major key role in the epidemiology of toxoplasmosis, rapid and accurate diagnosis of infected cats has utmost importance. In this study, we developed a novel colorimetric loop mediated isothermal amplification (LAMP) assay detecting T. gondii RE gene and modified a previously developed colorimetric LAMP assay targeting B1 gene to detect T. gondii DNA in cat feces for the first time. The analytical sensitivity of colorimetric LAMP assays was determined using plasmid controls. The clinical sensitivities of both colorimetric LAMPs were determined using cat fecal DNA samples that were confirmed to be positive by two different real-time PCRs in our previous study. According to the results, analytical sensitivities of both assays were 1 copy plasmid/reaction. Using real-time PCR as a reference method, sensitivities of colorimetric LAMP assays targeting RE and B1 genes were 100% and 97.56% whereas specificities of both assays were 100%. Overall, the colorimetric LAMP RE assay developed in this study brings an advantage in the diagnosis of T. gondii in cat fecal samples since it has higher sensitivity, does not need for experienced personnel, and can be applied in basic laboratories or in the field.