Naser Nazari

Background: Leishmaniasis remains a major global health issue, with current treatments often limited by side effects, drug resistance, prolonged treatment durations, and high costs. These challenges have driven interest in alternative therapies. Objectives: The present study aimed to assess the in vitro and in vivo effects of pomegranate peel aqueous extract against Leishmania major. Methods: The study was a randomized, controlled, parallel-group trial with a 1:1:1:1:1 allocation ratio. Forty healthy female BALB/c mice (6 - 8 weeks old, 40 - 50 g) were subcutaneously infected with 2 × 106 L. major promastigotes. Once lesions developed, mice were randomly assigned to five groups (n = 8 per group). Randomization and blinding of outcome assessors were employed to minimize bias. Four groups received topical applications of pomegranate peel aqueous extract at concentrations of 2.5%, 5%, 20%, or 40%, while the control group received physiological serum. Treatments were applied directly to the lesion using a sterile cotton swab, twice daily for 30 days. The primary outcome was the change in lesion size, measured with a digital caliper at baseline and day 30. Secondary outcomes included the in vitro viability of L. major promastigotes after 72-hour exposure to extract concentrations (0.1 - 0.9 mg/mL), evaluated by trypan blue exclusion assay using a Neubauer chamber. Paired t-tests compared pre- and post-treatment lesion sizes within groups, and one-way ANOVA with post-hoc analysis compared between groups. A P ≤ 0.05 was considered statistically significant. Results: The extract inhibited L. major promastigote growth at concentrations between 0.3 and 0.9 mg/mL in vitro. In vivo, lesion size reductions were dose-dependent. Treatment with 20% extract led to a 27% reduction in lesion size (from 0.37 ± 0.03 cm to 0.27 ± 0.03 cm; mean difference = 0.10 cm, 95% CI: 0.07 - 0.13 cm, P < 0.05). The most significant effect was observed with 40% extract, resulting in an 84.7% reduction in lesion size (from 4.90 ± 0.75 cm to 0.75 ± 0.11 cm; mean difference = 4.15 cm, 95% CI: 3.85 - 4.45 cm, P < 0.05). Lower concentrations (2.5% and 5%) showed no statistically significant difference compared to the control group (P > 0.05). Conclusions: Pomegranate peel aqueous extract appears to be a promising treatment for cutaneous leishmaniasis (CL). However, the study has several limitations, including the absence of a standard anti-leishmanial drug as a positive control, potential imprecision in dosing due to percentage-based concentrations, and the use of a crude extract without quantification of active compounds. Further studies should compare extract types and identify specific therapeutic components.

Human toxoplasmosis is a global public health concern and a commercial vaccine is still lacking. The present in silico study was done to design a novel vaccine candidate using tachyzoite-specific SAG1-realted sequence (SRS) proteins. Overlapping B-cell and strictly-chosen human MHC-I binding epitopes were predicted and connected together using appropriate spacers. Moreover, a TLR4 agonist, human high mobility group box protein 1 (HMGB1), and His-tag were added to the N- and C-terminus of the vaccine sequence. The final vaccine had 442 residues and a molecular weight of 47.71 kDa. Physico-chemical evaluation showed a soluble, highly antigenic and non-allergen protein, with coils and helices as secondary structures. The vaccine 3D model was predicted by ITASSER server, subsequently refined and was shown to possess significant interactions with human TLR4. As well, potent stimulation of cellular and humoral immunity was demonstrated upon chimeric vaccine injection. Finally, the outputs showed that this vaccine model possesses top antigenicity, which could provoke significant cell-mediated immune profile including IFN-γ, and can be utilized towards prophylactic purposes.

Visceral leishmaniasis is a life‐threatening infectious disease worldwide. Extensive experiments have been done to introduce potential vaccine candidates to combat leishmaniasis. The present study was done to evaluate Leishmania donovani hydrophilic acylated surface protein B1 as a potential vaccine candidate using in silico methods. For this aim, server‐based predictions were performed regarding physicochemical characteristics, solubility, antigenicity, allergenicity, signal peptide, transmembrane domain, and posttranslational modifications (PTMs). Also, secondary and tertiary structures were predicted using NetSurfP‐3.0 and I‐TASSER, respectively. The 3D model was further subjected to refinement and validation, and promising B‐cell, cytotoxic T‐lymphocyte (CTL; human, dog), and helper T‐lymphocyte (HTL; human) epitopes were predicted. The protein had a molecular weight of 42.19 kDa, with high solubility (0.749), stability (instability index: 21.34), and hydrophilicity (GRAVY: ‐2.322). No signal peptide or transmembrane domain was predicted, and the most abundant PTMs were phosphorylation, O‐glycosylation, and acetylation. Many coils and disordered regions existed in the secondary structure analysis, and the tertiary model had a good confidence score (‐0.79). Next, the ProSA‐web and PROCHECK tools showed adequate improvements in the refined model compared to the crude model. Only 4 shared B‐cell epitopes among three web servers (ABCpred, BepiPred 2.0, and SVMTriP) were shown to be antigenic, nonallergenic, and with good water solubility. Also, five potent CTL epitopes in dogs and five in humans were predicted. Notably, two HTL epitopes were found to be potential IFN‐γ inducers. In conclusion, our results demonstrated several immunogenic epitopes in this protein, which could be directed towards multiepitope vaccine design.

Background Sarcocystis species are diverse apicomplexan parasites, though only two zoonotic species (S. hominis and S. heydorni) circulate between cattle and humans. Due to the importance of cattle in the human food chain and to prevent the consequences of parasitosis in humans, the first global systematic review and meta-analysis on molecular epidemiology, species distribution, and zoonotic significance of Sarcocystis infection in cattle was performed. Methods For this aim, four international English databases (PubMed, Scopus, Google Scholar, and Web of Science) were systematically searched till 20th September 2021, and random-effect models were drawn to calculate total estimates and their 95% confidence intervals (CIs). Results Finally, 44 papers from 21 countries were qualified for this review which examined 8526 cattle regarding Sarcocystis infection, rendering a total prevalence of 62.7% (95% CI 53–71.5%). Globally, 12 Sarcocystis spp. have been reported from cattle, including S. cruzi, S. hominis, S. hirsuta, S. rommeli, S. heydorni, S. bovifelis, S. bovini, S. sinensis, S. gigantea, S. fusiformis, S. hjorti and S. tenella. Among them, S. cruzi (37 studies), S. hominis (22 studies) and S. hirsuta (19 studies) were the 3 most common species, with 76.4% (95% CI 64.8–85%), 30.2% (95% CI 19.3–44%) and 8.7% (95% CI 3.8–18.6%), respectively. However, molecular identification was not performed in 48.4% (95% CI 27.3–70.1%) of the positive samples. Conclusion Despite the zoonotic significance of Sarcocystis spp., particularly S. hominis, the epidemiology and distribution of Sarcocystis infection in cattle remains unclear and demands more extensive researches around the world.

Background and Aim This study aimed to investigate the clinical and laboratory characteristics of hydatid cyst disease in patients admitted to Imam Reza, Imam Hossein, and Bisotoon hospitals in Kermanshah City, Iran, from 2014 to 2018. Methods & Materials In this descriptive study, we studied and analyzed the medical records of 350 patients with hydatid cysts who were hospitalized and underwent surgical treatment in Kermanshah hospitals from 2014 to 2018. Also, the demographic characteristics of patients, including gender, age, occupation, place of residence, and affected organs, were extracted and recorded. Ethical Considerations The Ethics Committee of Kermanshah University of Medical Sciences approved this study (Code: IR.KUMS.REC.1398.366). Results In total, 350 patients with a Mean±SD age of 43.2±19.7 in women and 43.3±20.7 in men were admitted with a diagnosis of hydatid cyst. The highest and lowest prevalence of hydatid cysts were in the age groups of 41-50 (16.5%) years and 1-10 (4.8%) years, respectively. The prevalence was 181 (51.7%) in women and 169 (48.2%) in men. The organs mostly infected with hydatid cysts were the liver, lung, and brain, with 241 cases (68.8%), 55 (15.7%), and 24 (6.8%), respectively. Conclusion This study shows that hydatid cyst disease is an important disease, especially in rural areas of Kermanshah Province, Iran.

Neospora caninum is an economically significant parasite among livestock, particularly in dairy cattle herds, causing storm abortions. Vaccination seems necessary to limit the infection and its harsh consequences. This is the first steps towards developing a multiepitope vaccine candidate against N. caninum using in silico approaches. High‐ranked mouse MHC‐binding and shared linear B‐cell epitopes from six proteins (SRS2, MIC3, MIC6, GRA1, IMP‐1, and profilin) as well as IFN‐γ‐inducing epitopes (from SAG1) were predicted, screened, and connected together through appropriate linkers. Finally, RS‐09 protein (TLR4 agonist) and histidine tag were added to N‐ and C‐terminal of the vaccine sequence, yielding 486 residues in length. Physicochemical properties showed a stable (instability index: 27.23), highly soluble, antigenic (VaxiJen score: 0.9554), and nonallergenic candidate. Secondary structure of the multiepitope protein included 58.85% random coil, 20.99% extended strand, and 20.16% alpha helix. Also, the tertiary structure was predicted, and further analyses validated a stable interaction between the vaccine model and mouse TLR4 (binding score: ‐1261.6). Virtual simulation of immune profile demonstrated potently stimulated humoral (IgG+IgM) and cell‐mediated (IFN‐γ) responses upon multiepitope vaccine injection. Altogether, a potentially immunogenic vaccine candidate was developed using several N. caninum proteins, with the capability to elicit IFN‐γ upsurge and other components of cellular immunity, and can be used in prophylactic purposes against neosporosis.

Prevention of neosporosis is advantageous for cattle health and productivity. Previously, several vaccine candidates were nominated for vaccination against Neospora caninum. This study was premised on in silico evaluation of N. caninum IMP‐1 in order to determine its physicochemical features and immunogenic epitopes. We employed a wide array of network‐based tools for the prediction of antigenicity, allergenicity, solubility, posttranslational modification (PTM) sites, physicochemical properties, transmembrane domains and signal peptide, secondary and tertiary structures, and intrinsically disordered regions. Also, prediction and screening of potential continuous B cell peptides and those epitopes having stringent affinity to couple with mouse major histocompatibility complex (MHC) and cytotoxic T lymphocyte (CTL) receptors were accomplished. The protein had 393 residues with a molecular weight of 42.71 kDa, representing aliphatic index of 85.83 (thermotolerant) and GRAVY score of ‐0.447 (hydrophilic). There were 47 PTM sites without a signal peptide in the sequence. Secondary structure comprised mostly of extended strand and helices, followed by coils. The Ramachandran plot of the refined model showed 90.1%, 9.9%, 0.0%, and 0.0% residues in the favored, additional allowed, generously allowed, and disallowed regions, correspondingly. Additionally, various potential B cell (linear and conformational), CTL, and MHC binding epitopes were predicted for N. caninum IMP‐1. The findings of the present study could be further directed for next‐generation vaccine design against neosporosis.

Aim: The prevalence of cutaneous leishmaniasis (CL) is growing in Iran, and new sources of the disease have been found in the country. The purpose of this study was to describe the epidemiology of CL in Qasr-e Shirin County, Kermanshah Province, West of Iran. Qasr-e Shirin is located near the Iran-Iraq border, and several million pilgrims pass through this area to Iraq every year. Materials and Methods: A descriptive cross-sectional study was carried out for active case detection from April 1, 2014, to March 31, 2015. All individuals with suspicious lesions were identified through house-to-house visits in the aforementioned area. A questionnaire was filled out for everyone with clinical suspicion of CL. A diagnosis was made based on parasitological examination of the lesion exudate. The annual incidence and prevalence of CL were estimated. Results: In total, 5277 individuals were randomly screened for the presence of active lesions or scars suspected of CL. The overall prevalence of CL in the County was 4.8% (60 cases had active lesions and 194 cases had scars). All age groups were affected, but the highest incidence of CL was seen in the age group 20-29 years (1.9%). The incidence and prevalence were higher in women than men (5.5% vs. 4%). Most cases (45.6%) had a single lesion or scar and 44 (17.4%) patients had five or more lesions or scars. Conclusion: The incidence of CL in Qasr-e Shirin County is high. There is a need for further studies on reservoirs and vectors of leishmaniasis in this area. The results of such studies help to perform more efficient and timely spraying against phlebotomine sandfly or detect and decrease the animal reservoir population.