Effects of Zinc Supplementation on Inflammatory Status and Nonalcoholic Steatohepatitis in Overweight or Obese Children: a Randomized Clinical Trial.

The purpose of the present clinical trial was to determine the impact of zinc supplementation on serum liver enzymes, steatosis severity, lipid profile, and inflammatory status in overweight or obese children with nonalcoholic steatohepatitis (NASH). This randomized controlled trial was conducted by enrolling 60 children with NASH, aged 10-18 years old. The participants were randomly assigned to two groups that received either 30 mg/day of elemental zinc or placebo for 16 weeks. The severity of liver steatosis was evaluated using liver ultrasonography. Fasting blood samples were collected from each patient at the beginning and after 16 weeks of intervention to measure biochemical parameters. Following a 16-week intervention, zinc supplementation compared with placebo significantly decreased serum alanine aminotransferase (ALT) concentrations and high-sensitivity C-reactive protein and considerably enhanced HDL-cholesterol values. However, zinc intake had no considerable impact on aspartate aminotransferase, the severity of liver steatosis, anthropometric parameters, and other lipid indices versus the placebo group. Overall, zinc supplementation showed a promising impact on serum ALT, HDL-cholesterol, and inflammatory status in overweight or obese children suffering from NASH. Zinc supplementation may be a new strategy for the amelioration of NASH in overweight or obese children. This trial has been registered on the Iranian website for registration of clinical trials with the special ID of IRCT20200531047614N1 ( https://www.irct.ir/trial/48543 ).

Antimalarial effects of the hydroalcoholic extract of Allium paradoxum in vitro and in vivo.

Malaria still is the most fatal parasitic disease affecting 50% of the world's population. Although annual deaths attributed to malaria has reduced, crucial importance of its prevention and treatment remains a priority for health care systems and researchers. The worldwide increase in resistance to most common antimalarial drugs such as chloroquine, their unpleasant side effects and low efficiencies persuade researchers to prioritize finding alternative drugs including herbal medication from plant roots. The present study aimed to examine in vitro and in vivo effects of hydroalcoholic extract of herbal medicinal plant, Allium paradoxum, on growth rate in Plasmodium falciparum and Plasmodium berghei. The cytotoxicity assay was performed for hydroalcoholic extract of A. paradoxum. The 3D7 strain of P. falciparum was cultured. The IC50 assay and enzymatic activity of lactate dehydrogenase were performed. BALB/c mice were infected with P. berghei in vivo. Toxicity and histopathological changes in the tissues of liver and kidney were also examined. The highest efficacy of A. paradoxum extract was observed at 80 µg/mL in P. falciparum culture resulting in 60.43% growth inhibition compared to control groups. The significantly highest parasite growth inhibition with 88.71% was seen in the mice infected with P. berghei when administered with 400 mg/kg extract compared to control groups. No significant changes in the liver and kidney cells were observed between experimental and control groups. The study showed that A. paradoxum extract exhibited significant antimalarial properties in vitro on P. falciparum and in vivo in mice infected with P. berghei. There was no significant toxicity in the liver and kidney of the treated mice.

Identification of Leishmania species by high-resolution melting analysis in newly emerged foci in Sabzevar, northeast of Iran.

Cutaneous leishmaniasis (CL) is a zoonotic disease with 1 to 1.5 million annual incidences. Microscopic examination of the Giemsa stained slides is the most common diagnostic method for CL. However, this method cannot distinguish leishmania species. Hence the present study was conducted to identify leishmania species by high-resolution melting (HRM) analysis in the newly emerged foci of CL in Sabzevar, northeast of Iran. In this cross-sectional study, fifty patients with suspicious cutaneous lesions referring to the designated health center for diagnosis and treatment of CL in Sabzevar during 2017-2018, were recruited. All collected samples and prepared slides were stained for microscopic examination and then undergone HRM real-time PCR (HRM-PCR) assay to identify species of Leishmania parasites. The results of HRM-PCR technique showed that Leishmania major (L. major) was the dominant causative parasite in the newly emerged foci whereas L. tropica (L. tropica) was positive only in two patients. This was the first time that 7SL RNA-HRM-PCR assay was performed to precisely identify leishmania parasites in the northeast of Iran. We proved the newfound foci in which both L. major and L. tropica were present. In contrast to the recent studies which identified only L. major in the region, we showed that L. tropica was still present.

Evaluation of Glutathione Bioactivity on Toxoplasma Gondii in BALB/c Mice Post Impact of Selenium and Calcium Supplementation.

BACKGROUND: Studies have shown that selenium is an essential component of glutathione as an important antioxidant to reduce oxidative stress and inhibit intracellular parasites' growth. In contrast, calcium in the cytosol of such parasites plays a key role in the entry of the parasite into the host cell and its primary motility. AIMS AND OBJECTIVES: The present study was designed to evaluate and compare glutathione peroxidase bioactivity effects post administration of selenium and calcium in BALB/c mice infected by Toxoplasma gondii. METHODS: Sixty BALB/c mice susceptible to T. gondii were randomly divided into twelve groups of case and control groups. There were six control groups including two positive controls infected only with the parasites either 104 or 5×104, non-infected and untreated groups. Treated controls received only calcium, selenium, or both respectively. Case groups were infected with 104 or 5×104 parasites. While each set of three case groups separately received minerals alone or together. Mice were orally fed with 200 µg selenium, 50 µg calcium or their combination for 7 days. Mice were infected by parasite's tachyzoites. Sera of mice were kept and the peritoneal macrophages were isolated for counting tachyzoites during infection. RESULTS: The results showed that selenium unlike calcium was significantly effective in reducing Toxoplasma tachyzoites compared to control groups. Moreover, glutathione peroxidase [GPX] activity was elevated in mice treated with selenium and vice versa decreased in mice treated with calcium. CONCLUSION: Administration of selenium unlike calcium reduced Toxoplasma tachyzoites proliferation by elevating bioactivity of selenium-dependent detoxification enzyme, GPX.

In Vitro Antiprotozoal Effects of Nano-chitosan on Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis.

BACKGROUND: Treatment of parasitic infections with conventional drugs is associated with high toxicity, and undesirable side effects require cogent substitutions. Nanotechnology has provided novel approaches to synthesize nano-drugs to improve efficient antipathetic treatment. PURPOSE: Nano-chitosan as a nontoxic antimicrobial agent was examined against three most prevalent protozoa in humans, Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis. METHODS: Chitosan extracted from Penicillium fungi was converted to nanoparticles to maximize its therapeutic properties. Safety of nano-chitosan was examined by determining its hemolytic property and toxicity on PC12 cells. The studied parasites were identified with RFLP-PCR and cultivation in relevant media. Characteristics of nano-chitosan as an useful and valuable curative compound was evaluated by FTIR, DLS and SEM. Dose dependent anti-parasitic effect of nano-chitosan was evaluated. RESULTS: The highest anti-parasitic activity of the nano-chitosan was observed at 50 µg/mL by which growth rates of cultivated P. falciparum, T. vaginalis and G. lamblia were inhibited by 59.5%, 99.4%, and 31.3%, respectively. The study demonstrated that nano-chitosan with the least toxicity, low side effects, and substantial efficacy deserved to be considered as an anti-parasitic nano-compound. CONCLUSION: Nano-chitosan significantly inhibited protozoan growth in vitro promising to explore its use to combat parasitic infections. Further investigations covering extended sample size, in vivo experiments and optimizing the concentration used may lead to efficient treatment of protozoan diseases.

Status of humoral and cellular immune markers in human T-cell lymphotropic virus type 1 (HTLV-1) asymptomatic carriers in northeastern Iran, Mashhad.

It is estimated that about 10-20 million peoples are infected with human T-cell leukemia virus type 1 (HTLV-1) around the world and suffered from HTLV-related diseases. The present study was aimed to evaluate the cellular immunity, T-cell activation, humoral immunity, and inflammatory response hallmarks which affect HTLV-1-associated disease progression. A total of 78 participants were included in the study, comprising 39 HTLV-1 asymptomatic careers (ACs) and 39 healthy controls. The HTLV-proviral load (PVL) was determined via real-time PCR technique, and anti-HTLV antibody, sIL2R, sCD30, Neoptrin, hs-CRP, IgE, anti-VCA, anti-EBNA, and anti-EA were assessed by ELISA method. Mean PVL in ACs was 352.7 ± 418.7 copies/104 PBMCs. A significant higher level of sIL-2R was observed in ACs (P < 0.0001). Anti-VCA antibody titer in ACs and healthy controls was 80.72 ± 105.95 and 156.05 ± 130.71, respectively (P = 0.007). Intriguingly, suppression in ACs immune response was not observed. Resultantly, HTLV-1 infection has no effect on the humoral immune response in ACs but greater T-cell activation and function cellular responses were detected. Finally, more studies on various immune markers in adult T-cell leukemia/lymphoma (ATLL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients are greatly needed to illuminate the association of ACs' immune status with the development of the related diseases.

Protective effects of Vitex pseudo-negundo leaves on diabetic-induced nephropathy in rats.

Objectives The belief of therapeutic effects of herbal remedies in diseases such as diabetes is rooted in medical history. The present study evaluated protective efficacy of the hydroalcoholic extract of Vitex pseudo-negundo leaves (VLHE) on the renal disorders in streptozotocin-induced diabetic rats. Methods Fifty Wistar male rats were recruited and divided into five groups of 10, including healthy controls and diabetic controls: three diabetic groups of which first group was treated with glibenclamide, and two groups treated with 250 and 500 mg/kg of VLHE, respectively, for six weeks. Renal biochemical tests and tissue histopathological evaluation were performed and the antioxidant status was examined. Results There were significant decreases in superoxide dismutase and glutathione peroxidase activities and increases in malondialdehyde levels in renal tissue of diabetic groups compared with healthy controls. In the VLHE-treated rats, fasting blood sugar, blood urea nitrogen and creatinine were declined, serum albumin elevated, kidney weight lowered, lipid peroxidation and reinforcement of the activities of antioxidant enzymes decreased compared with healthy groups. Histological assessments revealed that the vacuolar degeneration of tubules and shrinkage of glomeruli in VLHE-treated rats was decreased compared with diabetic rats. Conclusions The study suggested that administrating of VLHE in nephropathic rats ameliorated the disease by reduction of oxidative stress and increase in renal antioxidant enzyme activities.

Bovine herpesvirus type 5 in semen samples from bulls in Iran.

Bovine herpesvirus 5 (BoHV-5) is an important pathogen of the central nervous system and has already been described in the genital tract of cattle and in semen. This virus is responsible for sporadic epizootics of fatal meningoencephalitis of calves. The objective of the present study was the identification and characterization of BoHV-5 in semen samples from bulls for the first time in Iran. DNA was extracted from bull semen samples, and the glycoprotein D (gD) gene of BoHV-5 and also the thymidine kinase (tK) gene of bovine herpesvirus 1 (BoHV-1) were amplified by PCR assay. The results showed a high prevalence of BoHV-5 (73.2 %) and BoHV-1 (25.89 %) in Iranian bull semen samples. In addition, in order to identify and compare BoHV-5 isolated from Iranian bulls with other isolates from all over the world, the gD gene of this virus was cloned and sequenced. A BLAST search showed that the sequence of the gD gene of BoHV-5 from Iran was 99 % identical to other sequences in the GenBank database. The present study indicated that semen samples are important transmission sources of BoHV-5 virus in Iranian bulls.

Gentamicin-attenuated Leishmania infantum vaccine: protection of dogs against canine visceral leishmaniosis in endemic area of southeast of Iran.

An attenuated line of Leishmania infantum (L. infantum H-line) has been established by culturing promastigotes in vitro under gentamicin pressure. A vaccine trial was conducted using 103 naive dogs from a leishmaniosis non-endemic area (55 vaccinated and 48 unvaccinated) brought into an endemic area of southeast Iran. No local and/or general indications of disease were observed in the vaccinated dogs immediately after vaccination. The efficacy of the vaccine was evaluated after 24 months (4 sandfly transmission seasons) by serological, parasitological analyses and clinical examination. In western blot analysis of antibodies to L. infantum antigens, sera from 10 out of 31 (32.2%) unvaccinated dogs, but none of the sera from vaccinated dogs which were seropositive at >100, recognized the 21 kDa antigen of L. infantum wild-type (WT). Nine out of 31 (29%) unvaccinated dogs, but none of vaccinated dogs, were positive for the presence of Leishmania DNA. One out of 46 (2.2%) vaccinated dogs and 9 out of 31 (29%) unvaccinated dogs developed clinical signs of disease. These results suggest that gentamicin-attenuated L. infantum induced a significant and strong protective effect against canine visceral leishmaniosis in the endemic area.