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The use of venom fractions from the Iranian cobra could be useful adjunct treatments of malaria with chloroquine. A metabolomic investigation with 1HNMR spectroscopy was conducted on an effective fraction tested earlier using Plasmodium berghei as an experimental murine model. We sought to ascertain both safety and anti-parasitic effects of experimental therapies. After purification of the venom fractions, 25 mice were infected, then treated for 4 days with 0.2 ml of 5 mg/kg, 2.5 mg/kg and 1 mg/kg of the effective fraction, chloroquine, and a drug vehicle. An ED50 was obtained using Giemsa staining and real-time PCR analysis. The toxicity tests inspecting both liver and kidney tissues were performed. A clear inhibitory effect on parasitaemia was observed (with 75% inhibition with 5 mg/kg and 50% reduction when 2.5 mg/kg dosage used). ED50 obtained 2.5 mg/kg. The metabolomics were identified as differentiation of aminoacyl-t-RNA biosynthesis, valine, leucine, isoleucine biosynthesis and degradation pathways were observed. Upon therapeutic effects of cobra venom fraction, further optimization of dose-dependent response of pharmacokinetics would be worthwhile for further exploration in adjunct experimental venom therapies.

Background: Although a majority of patients with cutaneous leishmaniasis (CL) are healed with Glucantime chemotherapy, the increased drug resistance rate following its application is a concern. In this study, Naja naja oxiana crude venom of cobra snakes was used for the first time as an assembled combination of bioactive pharmaceutical components on Leishmania major (L. major) standard strain. Objective: To evaluate the efficacy of the Naja naja oxiana crude venom of Iranian cobra snakes on Leishmania major standard strain in vitro. Methods: Five concentrations (1.25, 2.5, 5, 10 and 20 μg/mL) of venom were added to Leishmania major cultures at 24, 48 and 72 h. The viability of the parasites and venom toxicity were assessed by MTT test. The apoptosis was determined by flow cytometry, while IC50 was determined by counting parasites compared to that of glucantime. Each test was conducted in triplicate. Results: After the exposure of the venom for 72 h, IC50 value was 0.36 μg/mL and 14.12 μg/mL for promastigotes and amastigotes, respectively. MTT valuation clarified 1.01% promastigotes viability. Furthermore, the results indicated that the venom of Naja naja oxiana induced early and late apoptosis in parasites. Conclusions: The venom of Naja naja oxiana revealed remarkable anti-leishmanial effects as a novel anti-parasitic alternative. Thus, the bioactive and effective fractions of this venom may be considered as anti-leishmanial candidates in future studies.

Objective: The purpose of this study is to evaluate the diagnostic value of nitrite test in comparison with urine culture (U/C) results in patients with Urinary tract infection (UTI) symptoms. Methods: This cross-sectional study was performed on 203 patients with UTI symptoms. Middle urine sample was taken into sterile plastic containers, and simultaneously the urine sample and the nitrite test were done by the use of urine dipstick test. Data were analyzed using SPSS software version 23. Results: The results of the urine culture test indicated that the highest excreted organisms in both sexes were E. coli (58.8%), Candida (17.6%), Klebsiella (8.8%), E. coli (MDR) (5.9%), Pseudomonas (2.9%), Enterococcus (2.9%) and Acinetobacter (2.9%). The results showed that there was a significant correlation between U/C and urine analysis (U/A) test results (P = 0.01), and in U/C positive results, U/A results were significantly positive for UTI. Other findings showed a significant relationship between the results of nitrite one and nitrite two tests (P = 0.001). There was a significant correlation between urinary, and nitrite1 levels (P = 0.04). Also, in this study, the sensitivity and specificity of diagnostic tests for U/A and nitrite 1 and 2 with U/C were calculated as the Golden Standard method. Conclusion: Overall, the results of this study showed that the negative nitrite test and urine dipstick test could be performed in emergency cases to prevent ectopic dysfunction and inadequate diagnosis. Also, it can reduce the unreasonable expenses for U/A.

We used TSA as a principal antigen of Leishmania major with adjuvants. The aim of this research was to assess the efficacy of protein vaccine in the presence of Montanide vis-a-vis chitosan nanoparticle. The expression of recombinant protein was confirmed with SDS (sodium dodecyl sulfate) page and Western blotting. A total of 36 BALB/c mice were divided into six groups (TSA/high chitosan, TSA/low chitosan, TSA/Montanide, high chitosan, low chitosan, and PBS groups) and subcutaneously immunized with 20 mg of vaccine at three time intervals on days 0, 14, and 28. The mice were challenged with parasites 21 days after the final immunization. For assaying lymphocyte proliferation, the Brdu test and for evaluation of IgG subclasses and cytokines, the ELISA method was carried out. No statistically noteworthy variation was found between the vaccinated groups before and after the infectious challenge or intervention in terms of IgG and IFN-γ values. Statistically, significant difference was seen among the vaccinated and control groups in terms of IgG and IFN-γ. In terms of IL-4 before and after intervention, no statistically substantial difference was noted in the vaccinated and control groups, whereas, IgG isotypes rose to a great extent prior to and following intervention in all vaccine-immunized groups as compared with control groups. The formulated recombinant TSA protein vaccine with adjuvants induced lymphocytes proliferation and increased cytokines and antibodies as compared with the control groups. Given their potency, the formulated candidate vaccines may be recommended for further studies.

Background: Bioethanol produced by fermentative microorganisms is regarded as an alternative to fossil fuel. Bioethanol to be used as a viable energy source must be produced cost-effectively by removing expense-intensive steps such as the enzymatic hydrolysis of substrate. Consolidated bioprocessing (CBP) is believed to be a practical solution combining saccharification and fermentation in a single step catalyzed by a microorganism. Bacillus subtills with innate ability to grow on a diversity of carbohydrates seems promising for affordable CBP bioethanol production using renewable plant biomass and wastes. Results: In this study, the genes encoding alcohol dehydrogenase from Z. mobilis (adhZ) and S. cerevisiae (adhS) were each used with Z. mobilis pyruvate decarboxylase gene (pdcZ) to create ethanologenic operons in a lactate-deficient (Δldh) B. subtilis resulting in NZ and NZS strains, respectively. The S. cerevisiae adhS caused significantly more ethanol production by NZS and therefore was used to make two other operons including one with double copies of both pdcZ and adhS and the other with a single pdcZ but double adhS genes expressed in N(ZS)2 and NZS2 strains, respectively. In addition, two fusion genes were constructed with pdcZ and adhS in alternate orientations and used for ethanol production by the harboring strains namely NZ:S and NS:Z, respectively. While the increase of gene dosage was not associated with elevated carbon flow for ethanol production, the fusion gene adhS:pdcZ resulted in more than two times increase of productivity by strain NS:Z as compared with NZS during 48 h fermentation. The CBP ethanol production by NZS and NS:Z using potatoes resulted in 16.3 g/L and 21.5 g/L ethanol during 96 h fermentation, respectively. Conclusion: In this study for the first time, Bacillus subtilis was successfully used for CBP ethanol production with S. cerevisiae alcohol dehydrogenase. The results of the study provide insights on the potentials of B. subtilis for affordable bioethanol production from inexpensive plant biomass and wastes. However, the potentials need to be improved by metabolic and process engineering for higher yields of ethanol production and plant biomass utilization.

Objective To evaluate the efficacy of some medicinal plants and systemic glucantime in a comparative manner against the causative agent of cutaneous leishmaniasis both in vitro and in BALB/c mice. Methods For in vivo testing, inbred mice were challenged with Leishmania major parasites and the resultant ulcers were treated with extract based-ointments applied topically two times per day for a period of 20 days. A group of 56 mice were randomly divided into 7 subgroups. The control group received the ointment void of extracts, whereas the reference group received glucantime only. The efficacy of treatments was evaluated by measuring ulcer diameter, parasite burden and NO production. Results Our results indicated that plant extract based-ointments were effective in reducing ulcer size and parasite burden in spleens, but their effects did not differ significantly from that of glucantime. The plant extracts tested in this study were able to increase NO production that helped parasite suppression. Conclusions Our findings indicate that the tested plant extracts are effective against Leishmania major both during in vitro and in vivo experiments, but further researches are required to recommend a potential plant extract as an alternative drug.

Background: Today, most patients with cutaneous leishmaniasis are treated with drugs such as glucantime; however, severe drug resistance to these drugs is found, in addition to recurrence, extensive drug complications, and secondary infections in many patients. Objectives: Therefore, due to the lack of sufficient information on this subject, this study was undertaken for the first time to investigate the antimicrobial activity of multifunctional lactoferrin as one of the most important bioactive compounds on amastigotes and promastigotes forms of Leishmania major in vitro. Methods: In this study, six concentrations of lactoferrin (2.5, 5, 10, 20, 40, and 80 µg/mL) were added to parasite cultures at specified times. The MTT was done. Leishmania major promastigotes were incorporated to J774 cells and then incubated for 72 hours. The results were compared with the results of glucantime, as a standard. In the end, flow cytometry was performed to investigate apoptosis. Results: After 24, 48, and 72 hours of incubation in the presence of various concentrations of lactoferrin, the number of amastigotes and promastigote parasites increased over time without any significant difference compared to the control group (P > 0.05), but it was significant compared to glucantime at the 80 µg/mL concentration (P < 0.05). Besides, lactoferrin showed very low toxicity effects on macrophages and stimulated the growth of both forms of L. major parasite. Conclusions: Our findings indicated that lactoferrin could not induce early and late apoptosis in both forms of L. major.

Objective: To evaluate immunoglobulin G (IgG) avidity as a useful and reliable technique in diagnosing toxoplasmosis in pregnant women referring to therapeutic centers of Alborz Province (Iran) in 2014, against two other tests, IgG and immunoglobulin G (IgM) antiToxoplasma. Methods: Serum samples (468 in total) were obtained from different therapeutic centers in Karaj City. ELISA method was used to test the anti-Toxoplasma avidity of IgG, IgM and IgG. The data were analyzed by descriptive statistical methods and Chi-square test (P < 0.05) using SPSS 17.0. Results: Anti-Toxoplasma tests of IgM and IgG were positive in 9 and 86 samples respectively. Also, a borderline IgM was detected in 2 suspected samples. In addition, among all positive and suspected samples, 79 cases indicated high titers of IgG avidity, 7 cases were of low titers and 1 case was of a borderline titer. The prevalence of anti-Toxoplasma antibodies was 20%. The sera which showed high avidity index was obtained from patients at chronic phase of infection (92%) while those which showed low avidity levels were from patients at acute toxoplasmosis (77.7%). Conclusions: This study clearly showed that acute and chronic phases of toxoplasmosis could be differentiated with the aid of IgG avidity test. This test may also assist in recognizing old and newly acquired infections.

BACKGROUND: Leishmaniasis is a parasitic disease induced by a protozoan from the genus Leishmania . No effective vaccine has yet been developed against the disease. AIM: In this work, two nano-vaccines, TSA recombinant plasmid and dendrimer and poly (methyl methacrylate) (PMMA) nanoparticles (as adjuvants), were designed and tested for their immunogenicity in BALB/c mice. METHODS: After the plasmid construction and preparation of adjuvants, three intramuscular injections of the nano-vaccines (100 µg) and the recombinant TSA protein (20 µg) were subcutaneously performed. Eventually, the challenged animals were infected with the parasites (1*10 6 promastigotes). After the last injections of the nano-vaccines, the responses of their antibody subclasses and cytokines were assessed via ELISA method before and after the challenge. RESULTS: This study revealed that the new nano-vaccines were strong and effective in inducing specific antibody and cellular responses and reducing the parasite burden in the spleen compared to the control groups of Leishmania major -infected BALB/c mice. CONCLUSION: Based on the results, we can suggest that the formulated vaccines are suitable candidates for further studies in the field of leishmaniasis control.

Leishmaniasis is a parasitic disease caused by several species of the genus Leishmania. Montanide ISA 70 is an adjuvant composed of a natural metabolizable oil and a very refined emulsifier from the manide monooleate family. The TSA (thiol-specific antioxidant) is a important antigen of Leishmania major. The purpose of this work was protein-vaccine efficacy as an protection and excellent candidate in the presence Montanide. The expression of recombinant protein was confirmed with SDS (sodium dodecyl sulfate) page and Western bloting. 48 BALB/c mice were divided into four groups (TSA/Freund,TSA/Alum + BCG, TSA/Montanide and PBS groups) and immunized with 20 μg of vaccine subcutaneously three times intervals on days 0, 14 and 28. The mice were challenged with parasite 21 days after final immunization. The lymphocyte proliferation was evaluated with Brdu method. Cytokines and also total antibody and subclasses were evaluated with ELISA method. The vaccine formulated with the recombinant TSA protein with Montanide induced lymphocytes proliferation cytokines and total antibody and subclasses as compared with the control group.