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Objective: Due to the spread of Coronavirus Disease 2019 (COVID-19) worldwide and the resulting mortality as well as the lack of definitive treatment for this disease, various non-invasive and invasive drugs and treatments have been used around the world for reducing the effects of this disease or accelerating the treatment process, but the positive and definite effect of none of these methods has been confirmed and they have been effective only in some cases. This study aims to investigate some clinical features and supportive therapies in deceased COVID-19 patients in Iran. Methods: This descriptive-analytical retrospective study was conducted on 100 Covid-19 patients died in Hajar Hospital in Tehran, Iran from February to September 2020. They were divided into two groups of with and without underlying diseases. After coordination and obtaining the necessary permission and informed consent, information was collected from the medical files of samples. Results: Mechanical ventilation, vasoconstrictor injection and blood products had been used as supportive therapies in deceased patients, where ventilation use (n=62, 62%) was significantly higher (P=0.01) compared to vasoconstrictor injection (n=28, 28%) and blood products (n=13, 13%). Most of deceased patients had an underlying diseases (79%). Hypertension, diabetes and heart problems were the most common underlying diseases. The disease severity, clinical manifestations, and mortality rate of deceased patients with cardiovascular diseases, diabetes and hypertension were significantly different compared to those with other underlying diseases (P= 0.01). The age and gender of deceased patients had no significant relationship with the use of any supportive therapies (P≥0.01), but had a significant relationship with the underlying diseases (P≤0.01). Conclusion: The treatment methods that are the definite needs of the patient and have more favorable effects on the recovery process have a high value in the treatment of COVID-19 patients. These patients need to use supportive therapies for purposeful and effective treatment. Mechanical ventilation is more important than injecting vasoconstrictors or using blood products.

Malaria is a parasitic lethal disease caused by Plasmodium protozoa. The resistance and drugs’ side effects have led to numerous researches for alternative suitable drugs with better efficiency and lower toxicity In the present study, we investigated in vivo antimalarial effects of G2 linear dendrimer-based nano-chloroquine. After the preparation of nano dendrimer G2, chloroquine loading was done. Determine the characterization of particles were specified by DLS, SLS and SEM. The LC–MS and FTIR were used for verifying the nano dendrimer G2 and the loading of chloroquine into the compound. The Solubility N-chloroquine and measurement of drug release rate were done. Antiplasmodial activity of N-chloroquine on BALB/c mice was performed by the microscope and enzymatic methods. At the end, In vivo toxicity of N-chloroquine on tissues was assayed. The RBC morphology and enzyme levels were identified. The results showed the synthesized N-chloroquine had suitable size and solubility. Highest inhibitory effect on Plasmodium parasitic growth was observed at 16 mg/kg dose of N-chloroquine, which eliminated 95% of the parasites (p > 0.05). ED50 is observed at 7.7 mg/kg of N-chloroquine dose. Biochemical findings showed the synthesized N-chloroquine was safer than chloroquine. The N-chloroquine no adverse effects were observed in examined tissues. Due to the better effect of the synthesized N-chloroquine on Plasmodium berghei in mice compared to chloroquine, this nanoparticle can be considered as an effective anti-plasmodium compound while more comprehensive research is recommended.

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 and Aims:: Due to the lack of an effective vaccine and complexity of the control measures against vectors and reservoir hosts, the control of leishmaniasis depends primarily on chemotherapy. This study was aimed to assess the snake venom, Naja naja oxiana fraction 11(NNOVF11) on Leishmania infantum and its broad mode of action. Methods:: A wide range of in vitro advanced assays including high-performance liquid chromatography (HPLC), MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5diphenyltetrazolium bromide; Thiazolyl blue), macrophage assays, quantitative real-time polymerase chain reaction (qPCR), flow cytometry and enzyme- linked immunosorbent assay (ELISA) on L. infantum promastigote and amastigote stages were used. IC50 values of L. infantum stages, CC50 value, and apoptosis were also analyzed. Results:: The NNOV-F11 demonstrated strong antileishmanial activity against L. infantum stages in a dose-dependent manner compared to the untreated control group. Interleukin (IL)-12, TNF-α, and iNOS genes expression as the indicators of T helper(h)1 response significantly increased; in contrast, the expression level of IL-10, as the representative of Th2 response significantly decreased (p < 0.001). Reactive oxygen species (ROS) detection showed a significant increase (p < 0.001) after treatment with different concentrations of NNOV-F11, unlike arginase (ARG) activity, which displayed a significant reduction (p < 0.001). Conclusion:: NNOV-F11 possessed a potent inhibitory effect on L. infantum stages with the multifunctional and broad mode of actions, which promoted the immunomodulatory role, induced ROS production, stimulated apoptotic–like mechanisms, and inhibited L-ARG activity, which collectively led to the parasite death. Further studies are crucial to assess the effect of the NNOV-F11 on animal models or clinical settings.

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.

The aim of this study was to explore the activity of Naja naja oxiana venom on Leishmania tropica and its modes of action. Different fractions of Naja naja oxiana venom (NNOV) were prepared and characterized by high-performance liquid chromatography. The superior component, fraction k (FK) was selected. The activity of the fraction was assessed using advanced assays. Interleukin (IL)-12, TNF-α and iNOS gene expression as the indicators of Th1 significantly increased. In contrast, the level of IL-10, as the marker of T helper 2 substantially decreased (p

As an important global disease, cutaneous leishmaniasis is associated with complications such as secondary infections and atrophic scars. The first line treatment with antimonials is expensive and reported to have serious side effects and enhance resistance development. The main objective of this study was to evaluate the effect of Cinnarizine on standard strains of Leishmania major because of paucity of information on this subject. Methods: In this experimental study, four concentrations of the drug (5, 10, 15 and 20 μg/ml) were added to Leishmania major cultures at 24, 48 and 72 hours intervals. MTT assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Half maximal inhibitory concentration (IC50) was ascertained by counting parasites. The inhibitory effect of the drug was compared with that of Glucantime. Flow-cytometry was performed to investigate apoptosis. Each test was repeated thrice. Results: The IC50 values of Cinnarizine after 72 hours were calculated to be 34.76 μg/ml and 23.73 μg/ml for promastigotes and amastigotes, respectively. The results of MTT assays showed 48 % promastigote viability after 72 hour-exposure to Cinnarizine at 20 μg/ml concentration. Programmed cell death in promastigote- and amastigote-infected macrophages was quantified to be 13.66 % and 98.7 %, respectively. Flow- cytometry analysis indicated that Cinnarizine induced early and late apoptosis in parasites. All treatments produced results which differed significantly from control group (P Conclusion: Cinnarizine showed low toxicity with anti-leishmanial and apoptosis effects on both promastigote and intracellular amastigote forms. Therefore, we may suggest further assessment on animal models of this drug as candidates for cutaneous leishmaniasis therapy.

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. Nano-chitosan as a nontoxic antimicrobial agent was examined against three most prevalent protozoa in humans, Plasmodium falciparum, Giardia lamblia and Trichomonas vaginalis. 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. 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. 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.

Visceral leishmaniasis (VL),a protozoan disease caused by Leishmania infantum is a major public health problem and cause of death among infants aged under 1 year and the elderly in endemic foci of Iran. The aim of this study is to determine the status of L.infantum infection in stray dogs from Meshkin-Shahr, a typical endemic area of VL in Iran.Sixty-eight randomly trapped stray dogs in Meshkin-Shahr area were tested for L. infantum infection using the direct agglutination test (DAT) from June to October 2016. The confirmation of seropositive samples was performed by Microscopic slides of spleen, culture and then PCR. The molecular methods performed by ITS1-PCR, RFLP-PCR and kDNA-PCR. The allof kDNA -PCR products were sequenced.Out of 68 examined stray dogs, 17 (25.0%) were positive for L. infantum by DAT (1:320 titers or higher). Parasite test showed that all of seropositive samples have amastigote forms in their spleens but only 3 out of them could be cultured. The kDNA-PCR confirmed all of seropositive samples but ITS1-PCR and RFLP-PCR only confirmed 3 out of 17 (17.6%) seropositive samples. The sequenced products showed 94% homology with L. infantum.The results showed a high prevalence of L. infantum infection in dogs in an endemic area of CVL and it provided key information for designing control programs against canine and human leishmaniasis.

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: Trichomonas vaginalis is a prevalent sexually transmitted infection cause trichomoniasis. In this study prevalence and genotype of Iranian isolates of T. vaginalis infected (dsRNA) viruses were evaluated by PCR-RFLP and obtained patterns were then confirmed by sequence analysis and genotype of these Iranian isolates confirmed again. Methods: Ten strains of T.vaginalis were collected from 1700 vaginal samples of women referred to hospitals associated with Iran University of Medical Sciences in Tehran, Iran during Feb 2016 to Jul 2017, evaluated in points of infection to T. vaginalis Virus (TVV-1) were used in a PCR-RFLP. All of ten isolates of T. vaginalis were examined by designed nested PCR for actin gene and then digestion patterns of three endonuclease enzymes of HindII, MseI and RsaI were evaluated and genotype of these isolates was defined. Results: By combination of fragments pattern of three enzymes of HindII, RsaI and MseI, three genotypes were found; six genotypes E, two genotypes G and two genotypes I. The most dominant genotypes were genotype E. Among four TVV infected isolates two genotype E, one genotype G and one genotype I were found, however among six uninfected T. vaginalis isolates to TVV-1, all of three genotypes were also found. Conclusion: Three genotypes E, G and I in T. vaginalis infected with dsRNA isolates were found, however, these three genotypes in T. vaginalis without virus were also observed. Further study is needed to evaluate genotypes of T. vaginalis, which infected virus in more great T. vaginalis population

The use of venom fractions from the Iranian cobra could be useful adjunct treatments of malaria with chloroquine. A metabolomic investigation withWe 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.

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 IC(50) 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.

Background and Aim: There is a large amount of information on intestinal parasites in stray dogs and golden jackals (Canis aureus) in Mazandaran Province, Iran. However, there is little information about foxes and wolves, which have a potential role in the spread of dangerous parasitic diseases, such as echinococcosis and toxocariasis. The aim of the present study was to identify the genus or species of parasitic worms in stool samples obtained from carnivores in Mazandaran Province, Iran, from August 2017 to April 2018. Materials and Methods: A total of 274 fecal samples were collected from carnivores, including dog, fox, wolf, and C. aureus in three areas of Mazandaran Province, Iran. All specimens were examined by centrifugal fecal flotation using a solution of Sheather's sugar to detect helminths eggs. Then, all samples were assessed using a light microscope. Data analysis was performed by SPSS version. 18 (Chicago, IL, USA). Results: In this study, seven genera of helminths were observed, including Ancylostoma, Uncinaria, Toxocara, Dipylidium, Toxascaris, Taenia, and Spirocerca. The prevalence of helminth infections was 97.7% (127 out of 130), 56.7% (51 out of 90), 51.4% (18 out of 35), and 52.6% (10 out of 19), among dogs, C. aureus, foxes, and wolves, respectively. The highest prevalence of Ancylostoma and Toxocara infections occurred in the eastern and central areas of the province (42.1% and 35.7%, respectively). Conclusion: Based on the results of this study, the infection with intestinal zoonotic helminths in carnivores was an important public health factor in Mazandaran. Therefore, these infections can be potentially harmful to humans and other animals.

Due to side-effects and inefficiency of the drugs used in malaria treatment, finding alternative medicine with less side-effects has attracted much attention. In this regard, in the present study, nanocomposite synthesized and its effects on the metabolites of P. falciparum were investigated. Subsequent to synthesis of nanocomposites, characterization was carried out using nuclear magnetic resonance (NMR), liquid chromatography-mass spectrometry (LC-MS), scanning electron microscopy, dynamic light scattering and Fourier-transform infrared tests. Solubility and drug release were measured and its toxicity on Vero cell was assessed using the MTT assay. The antiparasitic effect of the nanocomposite on the metabolites of P. falciparum was investigated by 1H NMR spectroscopy. Among synthesized nanocomposites, the average size of 239 nm showed suitable solubility in water as well as slow drug release. The MTT assay showed no toxicity for Vero cell lines. Concentrations of 2.5 μg mL-1 of nanocomposite eliminated 82.6% of the total parasites. The most effected metabolic cycles were glyoxylate and dicarboxylate metabolism. In this study, 1H NMR spectroscopy was used with untargeted metabolomics to study the effect of the nanocomposite on P. falciparum. Playing an essential role in understanding drug-target interactions and characterization of mechanism of action or resistance exhibited by novel antiprotozoal drugs, can be achieved by targeting metabolic using LC-MS.

Due to side-effects and inefficiency of the drugs used in malaria treatment, finding alternative medicine with less side-effects has attracted much attention. In this regard, in the present study, nanocomposite synthesized and its effects on the metabolites of P. falciparum were investigated. Subsequent to synthesis of nanocomposites, characterization was carried out using nuclear magnetic resonance (NMR), liquid chromatography-mass spectrometry (LC-MS), scanning electron microscopy, dynamic light scattering and Fourier-transform infrared tests. Solubility and drug release were measured and its toxicity on Vero cell was assessed using the MTT assay. The antiparasitic effect of the nanocomposite on the metabolites of P. falciparum was investigated by 1H NMR spectroscopy. Among synthesized nanocomposites, the average size of 239 nm showed suitable solubility in water as well as slow drug release. The MTT assay showed no toxicity for Vero cell lines. Concentrations of 2.5 μg mL−1 of nanocomposite eliminated 82.6% of the total parasites. The most effected metabolic cycles were glyoxylate and dicarboxylate metabolism. In this study, 1H NMR spectroscopy was used with untargeted metabolomics to study the effect of the nanocomposite on P. falciparum. Playing an essential role in understanding drug-target interactions and characterization of mechanism of action or resistance exhibited by novel antiprotozoal drugs, can be achieved by targeting metabolic using LC-MS.

Due to the lack of an effective vaccine and complexity of the control measures against vectors and reservoir hosts, the control of leishmaniasis depends primarily on chemotherapy. This study was aimed to assess the snake venom, Naja naja oxiana fraction 11(NNOVF11) on Leishmania infantum and its broad mode of action.A wide range of in vitro advanced assays including high-performance liquid chromatography (HPLC), MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5diphenyltetrazolium bromide; Thiazolyl blue), macrophage assays, quantitative real-time polymerase chain reaction (qPCR), flow cytometry and enzyme- linked immunosorbent assay (ELISA) on L. infantum promastigote and amastigote stages were used. ICThe NNOV-F11 demonstrated strong antileishmanial activity against L. infantum stages in a dose-dependent manner compared to the untreated control group. Interleukin (IL)-12, TNF-α, and iNOS genes expression as the indicators of T helper(h)1 response significantly increased; in contrast, the expression level of IL-10, as the representative of Th2 response significantly decreased (p0.001). Reactive oxygen species (ROS) detection showed a significant increase (p0.001) after treatment with different concentrations of NNOV-F11, unlike arginase (ARG) activity, which displayed a significant reduction (p0.001).NNOV-F11 possessed a potent inhibitory effect on L. infantum stages with the multifunctional and broad mode of actions, which promoted the immunomodulatory role, induced ROS production, stimulated apoptotic-like mechanisms, and inhibited L-ARG activity, which collectively led to the parasite death. Further studies are crucial to assess the effect of the NNOV-F11 on animal models or clinical settings.

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. Aim and Objective: 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.

Celiac disease is a genetic disease diagnosed to be associated with chronic intestinal inflammation. Endomysial and tissue transglutaminase antibodies are the best serum markers for celiac disease (CD) diagnosis. This research aimed to determine the levels of endomysial and tissue transglutaminase antibodies in patients with giardiasis. Forty cases of giardiasis were selected among the referees to Milad Hospital in Tehran, as well as a children’s hospital and some health centers in Karaj. Euro-immune IgA immunofluorescence and ELISA were utilized to measure their titers of endomysial and tissue transglutaminase antibodies, respectively. Twenty random serum samples of negative Giardia and 16 serum samples of positive anti-(tissue transglutaminase) tTG antibodies were evaluated by using ELISA and endomysium antibody through immunofluorescence techniques. Among the 40 cases of giardiasis, 16 positive endomysial and transglutaminase antibodies (40%) were detected. Sixteen positive samples of endomysial antibody (EMA) were also positive for anti-tTG, and 20 random negative samples were negative for EMA and anti-tTG. The chi-square test revealed a significant association between antibodies and giardiasis (P = 0.001). Atrophy of the intestinal villi can arise after giardia infection by mimicking the behavioral patterns of anti-EMA and anti-tTG antibodies. Due to low specificity, anti-gliadin antibody test is not helpful in detecting CD in patients with giardiasis. In the present study on anti-EMA and anti-tTG in patients with giardiasis, these antibodies were positive in giardiasis. Thus, the best and most reliable way to diagnose CD and treat giardiasis is intestinal biopsy as the gold standard method.

The present study showed that osmopriming or pretreatment with low H2O2 doses (2 mM) for 6 h alleviated salt-reduced seed germination. The NADPH oxidase activity was the main source, and superoxide dismutase (SOD) activity might be a secondary source of H2O2 generation during osmopriming or H2O2 pretreatment. Hematin pretreatment similar to osmopriming improved salt-reduced seed germination that was coincident with the enhancement of heme oxygenase (HO) activity. The semi-quantitative RT-PCR confirmed that osmopriming or H2O2 pretreatment was able to upregulate heme oxygenase HO-1 transcription, while the application of N,N-dimethyl thiourea (DMTU as trap of endogenous H2O2) and diphenyleneiodonium (DPI as inhibitor of NADPHox) not only blocked the upregulation of HO but also reversed the osmopriming-induced salt attenuation. The addition of CO-saturated aqueous rescued the inhibitory effect of DMTU and DPI on seed germination and α-amylase activity during osmopriming or H2O2 pretreatment, but H2O2 could not reverse the inhibitory effect of ZnPPIX (as HO inhibitor) or Hb (as CO scavenger) that indicates that the CO acts downstream of H2O2 in priming-driven salt acclimation. The antioxidant enzymes and proline synthesis were upregulated in roots of seedlings grown from primed seeds, and these responses were reversed by adding DMTU, ZnPPIX, and Hb during osmopriming. These findings for the first time suggest that H2O2 signaling and upregulation of heme oxygenase play a crucial role in priming-driven salt tolerance.

Objectives: Given the relatively high prevalence and emphasis on the pathogenicity of Blastocystis sp., especially in immunocompromised patients and other at risked populations like military personnel, this study was carried out to investigate the prevalence of Blastocystis sp. in the city of Karaj during 2014 - 2015. Methods: In this descriptive study, 2500 stool samples were screened for Blastocystis sp. using microscopically examinations and iodine-stained, wet-mount preparations and formalin-ether as a sedimentation method. Demographic information such as age, gender, and symptoms of patients, were recorded in the questionnaire. Three stool samples were collected from each patient in disposable plastic containers. Data were analyzed using chi-square, ANOVA, and t-test SPSS V. 24 software. Results: From the total of 2500 cases, 345 (13.8%) cases were positive for Blastocystis sp., 56.5% of patients were male, and 43.5% were female. A total of 33.3% of patients had a clinical symptom and 66.7% of patient had no clinical symptoms. Anorexia was the most common symptom in patients with Blastocystis sp. Conclusions: The use of appropriate diagnostic laboratory procedures in hospitals and medical centers can provide an effective and accurate diagnosis that will play an important role in the promotion of community health; considering some unspecific signs could help the health professionals to have correct diagnose.

Background: The first line treatment against cutaneous leishmaniasis is meglumine antimoniate. This drug is expensive and has serious side effects, including development of drug resistance. Objectives: In this research, because of paucity of information, the apoptotic and leishmanicidal effects of ketotifen and cromolyn sodium, as cell membrane stabilizer drugs, were investigated on standard strain of Leishmania major. Methods: In this experimental study, L. major parasites were first cultured in RPM1 1640 media, supplemented with 10% fetal bovine serum (FBS) and antibiotics at 24 ± 1oC. Drug concentrations of 5, 10, 15, and 20 µg/mL were then added to L. major culture at 24-, 48- and 72-hour intervals. The 3 - (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) tetrazolium assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Half maximal inhibitory concentration (IC50) were ascertained by counting the parasites. The inhibitory effect of the drugs were compared with that of glucantime. Flow cytometry was performed in the next step, to evaluate apoptosis. Each test was repeated three times. Results: IC50 values of ketotifen and cromolyn sodium after 72 hours were calculated to be 2.04 and 17.67 µg/mL for promastigotes and 0.12 and 14.79 µg/mL for amastigotes, respectively. The results of MTT assays showed 20% and 35% promastigote viability after 72 hours of exposure to ketotifen and cromolyn sodium at 20 µg/mL concentration. Apoptosis in ketotifen and cromolyn sodium was quantified to be 11.52% and 9.96% in promastigotes and 99.5% and 98.6% in amastigote-infected macrophages, respectively. The results indicated that the drugs induce early and late apoptosis in parasites. All treatments produced results, which differed significantly from the control groups (P < 0.05). Conclusions: Drugs used in this study, especially Ketotifen, showed lower toxicity yet similar anti-leishmanial effects on both forms, as cromolyn sodium did. It could be suggested that further investigations about the in vivo effects of these drugs, as candidates for cutaneous leishmaniasis treatment, are required.

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 and Objectives: Toxoplasmosis is a parasitic infectious disease. The occurrence of this disease brings about multiple complications resulting in abortion, death, or severe fetal complications. These complications may occur as a result of increased fibrinogen levels during pregnancy and increased serum factors associated with Toxoplasmosis. This study examined the plasma level of fibrinogen in the first trimester in mothers with acute toxoplasmosis for the first time. Methods: In this study, 205 pregnant women who were referred to various medical centers of Tehran during 2014 - 2015 were studied. The bloods were examined for the levels of IgG avidity, anti - toxoplasma IgM and IgG, and fibrinogen levels. SPSS software v. 22.0 was used to study the results. Data regarding their demographic characteristics were collected by a questionnaire. Results: Among a total of 205 patients, 41 cases had positive IgG titer, two had positive IgM titer (one low avidity), and five were borderline. Fibrinogen level was high in one case of positive IgM titer as well as low avidity and positive in 10 out of 41 cases with IgG titers. Conclusions: The results of the present study shows that increased levels of fibrinogen in pregnant women during the first trimester of pregnancy is not significantly associated with toxoplasmosis.

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.

The present study showed that osmopriming or pretreatment with low H

Leishmaniasis is an important disease in humans. Leishmania homologue of receptor for Activated C Kinase (LACK) and thiol specific antioxidant (TSA) as immuno-dominant antigens of Leishmania major are considered the most promising molecules for a DNA vaccine. We constructed a DNA cocktail, containing plasmids encoding LACK and TSA genes of Leishmania major and evaluated the immune response and survival rate in BALB/c mice. IgG and Interferon gamma values were noticeably increased in the immunized group with DNA cocktail vaccine, which were significantly higher than those in the single-gene vaccinated and control groups (p0.05) following the immunization and after challenging with Leishmania major. Interleukin 4 values were decreased in all immunized groups, but only in DNA vaccine cocktail and single-gene vaccination with pc-LACK there were statistical differences with control groups (p0.05). The immunized mice with the cocktail DNA vaccine presented a considerable reduction in diameter of lesion compared to other groups and a significant difference was observed (p0.05) in this regard. The survival time of the immunized mice with the cocktail DNA vaccine was significantly higher than that in the other groups (p0.05) after their being challenged with Leishmania major. The findings of this study indicated that the cocktail DNA vaccine increased the cellular response and survival rate and induced protection against infection with Leishmania in the mice.

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.

Leishmaniasis is a major infectious disease caused by protozoan parasites of the genus Leishmania. Despite of many efforts toward vaccine against Leishmania no effective vaccine has been approved yet. DNA vaccines can generate more powerful and broad immune responses than conventional vaccines. In order to increase immunity, the DNA vaccine has been supplemented with adjuvant. In this study a new nano-vaccine containing TSA recombinant plasmid and poly(methylmethacrylate) nanoparticles (act as adjuvant) was designed and its immunogenicity tested on BALB/c mouse. After three intramuscular injection of nano-vaccine (100 μg), the recombinant TSA protein (20 μg) was injected subcutaneously. Finally as a challenge animals were infected by Leishmania major. After the last injection of nano-vaccine, after protein booster injection, and also after challenge, cellular immune and antibody responses were evaluated by ELISA method. The findings of this study showed the new nano-vaccine was capable of induction both cytokines secretion and specific antibody responses, but predominant Th1 immune response characterized by IFN-γ production compared to control groups. Moreover, results revealed that nano-vaccine was effective in reducing parasite burden in the spleen of Leishmania major-infected BALB/c mice. Base on results, current candidate vaccine has potency for further studies.

Microsporidia infections occur in virtually all invertebrate and vertebrate hosts, including humans. The aim of this study is detection of microsporidiosis in various samples of Iranian immunosuppressed patients during 2011–2012 by molecular methods. The samples included stool samples from the healthy participants and samples from biological fluids of the patients according to consult of their physician and the site of infection. The sample size was determined as 258 for each group. Clinical and demographical data related to each participant was collected. DNA extraction and nested polymerase chain reaction were carried out on all the samples. In the control group, the rates of Encephalitozoon and Enterocytozoon infections were 5.3 and 4 % respectively higher in males and in the age range of 30–45, and all positive cases had gastrointestinal symptoms. In the patient group, most infection cases occurred in male patients and in the age range of 60 and above. Patients with microsporidiosis mostly had the symptoms of chronic diarrhea, vomiting, weight loss, dyspepsia, and malabsorption. In BAL samples from patients 2 % Encephalitozoon and 0.7 % Enterocytozoon, in the sampling bone marrow transplantation from patients 5.7 % Encephalitozoon, 1.43 % Enterocytozoon and from patients who underwent kidney transplantation 5.26 % Enterocytozoon were detected. The most cases of human microsporidiosis are associated with human immunodeficiency virus infection or other states of immunosuppression, particularly in organ transplant recipients; the result of this study confirms this claim.