The effects of Fasciola hepatica recombinant proteins (peroxiredoxin and cathepsin L1) on Crohn's disease experimental model.

The immunomodulatory potential of the excretory-secretory (E/S) proteins of the helminths has been shown in previous investigations. This study evaluated the effects of the recombinants and excretory-secretory proteins of the Fasciola hepatica on induced colitis in Balb/c mice. The F. hepatica Recombinant proteins, Cathepsin L1 and Peroxiredoxin, and E/S proteins were intraperitoneally injected into the three mice groups as the case groups, while the control groups received PBS. Colitis was induced in mice by intraluminal administration of the 2, 4, 6-Trinitrobenzenesulfonic acid solution (TNBS). After 8 h, the case groups received the second dosage of the treatments, and it was repeated 24 h later. The immunological, pathological, and macroscopic changes were evaluated 3 days after colitis induction. The macroscopic evaluation revealed significantly lower inflammatory scores in the mice treated with recombinant Peroxiredoxin (rPRX) and recombinant Cathepsin L1 (rCL1). Despite the macroscopic observation, the pathological finding was insignificant between the groups. IFN-γ secretion was significantly lower in splenocytes of the groups that received rPRX, rCL1, and E/S than the controls. IL-10 showed significantly higher levels in groups treated with rPRX and rCL1 than controls, whereas the level of IL-4 was not statistically significant. Excretory-secretory proteins of the F. hepatica showed immunomodulatory potency and the main effects observed in this study were through the reduction of inflammatory cytokine and inflammation manifestation as well as induction of anti-inflammatory cytokines.

The Therapeutic Effects of Curcumin-coated Gold Nanoparticle Against Leishmania Major Causative Agent of Zoonotic Cutaneous Leishmaniasis (ZCL): An In Vitro and In Vivo Study.

We synthesized and characterized curcumin-coated gold nanoparticles (Cur@AuNPs) and investigated their stability, cytotoxicity, leishmanicidal activity in in vitro and in in vivo experiments. Cur@AuNPs synthesized through a simple one-pot green chemistry technique. The in vitro leishmanicidal activity of curcumin-coated gold nanoparticles against extracellular promastigotes and intracellular amastigotes of protozoan parasite Leishmania major (L. major) was determined by applying the tetrazolium reduction colorimetric quantitative MTT technique. For in vivo assessment, the footpad lesion size and parasite burden in two infection site organs including lymph nodes and footpads of susceptible BALB/c mice infected with L. major were measured. Mice immune responses in all study groups were quantified by measuring the levels of gamma interferon (IFN-γ) and interleukin-4 (IL-4). Viability of Leishmania promastigotes significantly diminished with the inhibition in promastigotes growth (IC50) of 64.79 µg/mL and 29.89 µg/mL for 24 h and 48 h, respectively. In vitro nanoparticles treatment efficiently cleared the L. major amastigotes explanted in macrophages but had no harmful toxicity on the mice cells. In the in vivo condition, in the treated infected BALB/c mice the CL lesion size, Leishmania parasite burden, and IL-4 were decreased, while IFN-γ was significantly increased. The results suggest that Cur@AuNP was an effective compound against Leishmania parasite in vitro and in vivo, efficiently induced T-helper 1 (Th1) responses and augmented host cellular immune responses, and ending in a reduced Leishmania parasite burden. Therefore, it may be identified as a novel potential therapeutic approach for the local therapy of zoonotic CL treatment with high cure rates.

Blastocystis, urticaria, and skin disorders: review of the current evidences.

Blastocystis is one of the most common intestinal protozoan parasites worldwide, which is linked to cutaneous lesions and urticaria. In a setting of systematic review, the data on the association of Blastocystis infection with cutaneous lesions were searched in order to summarize the main clinical symptoms, diagnostic methods, treatment, and outcome of the patients. The search identified 28 eligible articles, including 12 cross-sectional studies and 16 case reports/case series (including 23 cases). A diverse spectrum of skin symptoms, mainly urticaria, rash, and itching, was reported from the studies. Of the 23 infected cases with the skin symptoms, gastrointestinal symptoms were reported from the 16 cases, whereas 7 cases with urticaria had asymptomatic infection. The most frequent subtypes were ST1, ST2, and ST3, respectively. Metronidazole, paromomycin, and tinidazole were the most prescribed drugs in patients with single Blastocystis infection. Notably, urticaria and other cutaneous symptoms of all treated patients were resolved after treatment. In conclusion, this study indicates that Blastocystis infection can be a neglected cause of urticaria and skin disorders. Since the treatment of Blastocystis infection is simple, screening and treatment of this infection should be considered in patients with urticaria and other skin disorders.

Sambucus ebulus extract stimulates cellular responses in cutaneous leishmaniasis.

This is the first study aiming to determine the therapeutic effects of the Sambucus ebulus aquatic extract as an antileishmanial herbal drug and evaluate the immune responses in Leishmania major major infected BALB/c mice. The antileishmanial activity of S ebulus aquatic extract was evaluated using MTT test as well as parasite rescue and transformation assay. Footpad swelling and parasite load of infected mice were measured by several techniques. The immune responses were evaluated by measuring the levels of IFN-γ, IL-4, nitric oxide and arginase. The results indicated that S. ebulus can significantly decrease L. major promastigotes and amastigotes viability, but it was not toxic to macrophages. The lesion size, parasite burden and the level of ARG decreased in the treated infected mice, while the IFN-γ-to-IL-4 ratio and the level of NO increased significantly. Altogether, the S. ebulus extract is an effective compound for killing Leishmania parasite without excessive toxicity to the host cells and can cure the CL by switching the host immune responses towards Th1 response. Thus, it may be a perfect therapeutic option for CL treatment.

Simulation of the potential impact of climate change on malaria incidence using artificial neural networks (ANNs).

Climate change can increase the spread of infectious diseases and public health concerns. Malaria is one of the endemic infectious diseases of Iran, whose transmission is strongly influenced by climatic conditions. The effect of climate change on malaria in the southeastern Iran from 2021 to 2050 was simulated by using artificial neural networks (ANNs). Gamma test (GT) and general circulation models (GCMs) were used to determine the best delay time and to generate the future climate model under two distinct scenarios (RCP2.6 and RCP8.5). To simulate the various impacts of climate change on malaria infection, ANNs were applied using daily collected data for 12 years (from 2003 to 2014). The future climate of the study area will be hotter by 2050. The simulation of malaria cases elucidated that there is an intense increasing trend in malaria cases under the RCP8.5 scenario until 2050, with the highest number of infections occurring in the warmer months. Rainfall and maximum temperature were identified as the most influential input variables. Optimum temperatures and increased rainfall provide a suitable environment for the transmission of parasites and cause an intense increase in the number of infection cases with a delay of approximately 90 days. ANNs were introduced as a practical tool for simulating the impact of climate change on the prevalence, geographic distribution, and biological activity of malaria and for estimating the future trend of the disease in order to adopt protective measures in endemic areas.

Co-infection of Phlebotomus papatasi (Diptera: Psychodidae) gut bacteria with Leishmania major exacerbates the pathological responses of BALB/c mice.

Clinical features and severity of the leishmaniasis is extremely intricate and depend on several factors, especially sand fly-derived products. Bacteria in the sand fly's gut are a perpetual companion of Leishmania parasites. However, consequences of the concomitance of these bacteria and Leishmania parasite outside the midgut environment have not been investigated in the infection process. Herein, a needle infection model was designed to mimic transmission by sand flies, to examine differences in the onset and progression of L. major infection initiated by inoculation with "low" or "high" doses of Enterobacter cloacae and Bacillus subtilis bacteria. The results showed an alteration in the local expression of pro- and anti-inflammatory cytokines in mice receiving different inoculations of bacteria. Simultaneous injection of two bacteria with Leishmania parasites in the low-dose group caused greater thickness of ear pinna and enhanced tissue chronic inflammatory cells, as well as resulted in multifold increase in the expression of IL-4 and IL-1ß and a decrease in the iNOS expression, without changing the L. major burden. Despite advances in scientific breakthroughs, scant survey has investigated the interaction between micro and macro levels of organization of leishmaniasis that ranges from the cellular to macro ecosystem levels, giving rise to the spread and persistence of the disease in a region. Our findings provide new insight into using the potential of the vector-derived microbiota in modulating the vertebrate immune system for the benefit of the host or recommend the use of appropriate antibiotics along with antileishmanial medicines.

Toxoplasmosis and symptoms severity in patients with COVID-19 in referral centers in Northern Iran.

Toxoplasmosis has been categorized as one of the long-lasting protozoan parasitic infections. It affects almost one-third of the world's population. In recent years, several documented studies have elucidated that infected individuals have a remarkably higher incidence of distinct health problems and show various adverse effects. In the PCR-positive COVID-19 patients in Gonbad-e-Kavus, Kalaleh, and Minoodasht counties in the northern part of Iran from June 2021 to December 2021, we sought to investigate any potential relationships between the severity of COVID-19 symptoms and acute and latent toxoplasmosis caused by Toxoplasma gondii (T. gondii). Whole blood samples of 161 COVID-19 patients with positive PCR. The samples were centrifuged to separate serum and screened for two important antibodies against T. gondii (IgM and IgG) by using ELISA kits for human anti-T. gondii IgM and IgG. Anti-T. gondii IgM and IgG antibodies were detected in 8/161 (5.0%) and 42/161 (26.1%) COVID-19 patients, respectively. No significant relationships were found between Toxoplasma IgM and IgG results with clinical signs, age, sex, contact with animals, comorbidities, and also the mortality rate of people with COVID-19. These findings showed that acute and latent toxoplasmosis infections are common among patients with COVID-19; however, no significant associations were found between toxoplasma infections and the symptoms of COVID-19. Therefore, toxoplasmosis is not considered a risk factor for COVID-19.

Corrigendum: Co-infection of Phlebotomus papatasi (Diptera: Psychodidae) gut bacteria with Leishmania major exacerbates the pathological responses of BALB/c mice.

[This corrects the article DOI: 10.3389/fcimb.2023.1115542.].

Anti-Leishmanial Effects of a Novel Biocompatible Non-Invasive Nanofibers Containing Royal Jelly and Propolis against Iranian Strain of Leishmania major (MRHO/IR/75/ER): an In-Vitro Study.

Background: Current medications especially the pentavalent antimonial compounds have been used as the first line treatment of cutaneous leishmaniasis (CL), but they have limitations due to serious side effects such as drug resistance, cardio and nephrotoxicity, and high costs. Hence, the demand to find more usable drugs is evident. Synthesis and development of natural, effective, biocompatible, and harmless compounds against Leishmania major is the principal priority of this study. Methods: By electrospinning method, a new type of nanofiber were synthesized from royal jelly and propolis with different ratios. Nanofibers were characterized by Scanning Electron Microscope (SEM), Transmission Electron Microscopy (TEM), Thermogravimetric Analysis (TGA), Contact angle, and Fourier-transform infrared spectroscopy (FTIR). The Half-maximal inhibitory concentration (IC50), Half-maximal effective concentration (EC50) and the 50% cytotoxic concentration (CC50) for different concentrations of nanofibers were determined using quantitative calorimetric methods. Inductively coupled plasma-optical emission spectrometry (ICP-OES) and flow cytometry were performed as complementary tests. Results: The results showed that the proposed formulas provide a new achievement that, despite the significant killing activity on L. major, has negligible cytotoxicity on the host cells. Royal jelly nanofibers have significantly shown the best 72 hours results (IC50= 35 µg/ml and EC50=16.4 µg/ml) and the least cytotoxicity. Conclusion: This study presents a great challenge to introduce a new low-cost treatment method for CL, accelerate wound healing, and reduce scarring with minimal side effects and biocompatible materials. Royal jelly and propolis nanofibers significantly inhibit the growth of L. major in-vitro.

Immunogenic properties of empty pcDNA3 plasmid against zoonotic cutaneous leishmaniasis in mice.

BACKGROUND: Leishmania (L) parasite, the causative agent of zoonotic cutaneous leishmaniasis (ZCL), effectively stimulates the mammalian cells to mount strong humoral responses by enhancing T-helper-2 (Th2)-associated cytokines for its survival. The best strategy to decrease the intensity of infection in the host is induction of cellular immunity. METHODS: We evaluated the effects of the empty bacterial pcDNA3 plasmid on mice infected with L. major and quantified the immune mediators including IFN-γ, IL-4, IL-10, IgG2a, IgG1, arginase activity and nitric oxide (NO) in the mice. Moreover, the footpad lesion size and parasite load were assessed. RESULTS: We observed that pcDNA3 could modulate the immune responses in favor of host cells and decrease the disease severity. Th2- associated mediators, including arginase, IL-4, and IL-10 are downregulated, while cellular responses are upregulated in line with an increase in the levels of nitric oxide (NO) and interfero-gamma (IFN-γ). Interestingly, pcDNA3 induced specific Th1-associated antibodies, IgG2a isotype; however, it suppressed the production of humoral IgG1. The stimulation of the immune response by the empty pcDNA3 is able to shift the immune function to predominant cellular responses caused by Th1, and it had a positive effect on the treatment of zoonotic cutaneous leishmaniasis (ZCL). CONCLUSIONS: Altogether, we introduced the pcDNA3 as a potential interfering factor in the modulation of the immune system against ZCL. Since this vector has been widely used as a control group in different studies, we suggest that the potential function of the empty vector should be deeply assessed, as it exerts anti-parasitic effects on mice infected with L. major.

Effects of negative air ions (NAIs) on Leishmania major: A novel tool for treatment of zoonotic cutaneous leishmaniasis (ZCL).

BACKGROUND: Cutaneous leishmaniasis (CL) is a Neglected Tropical Disease (NTD) that causes high morbidity in the tropics and sub-tropics. Despite the remarkable advancements in the treatment of CL, the available therapeutics are far from ideal and also cause serious adverse side effects. Negative air ions (NAIs) generators are widely available for domestic and industrial uses. Several studies have reported on positive effects of NAIs therapy on human health as a non-pharmaceutical treatment for respiratory disease, allergy, or stress-related health conditions, including infectious diseases. To our knowledge, no studies have examined the effectiveness of the NAIs therapy against Leishmania parasites. The aims of this study were to investigate the effect of NAIs therapy on Leishmania major (L. major) the causative agent of CL in in vitro and in a murine model. METHODOLOGY/PRINCIPAL FINDINGS: In vitro anti-leishmanial effects of NAIs therapy were measured by parasitological methods. NAIs therapy was assessed in vivo in L. major infected BALB/c mice by measuring the footpad (FP) lesion size and parasite load using metric caliper tool and qPCR, respectively. Immune responses in treated and non-treated mice were assessed by measuring the levels of IFN-γ, IL-4, NO and arginase activity. In vitro NAIs therapy significantly decreased the viability of Leishmania promastigotes and of amastigotes cultured in macrophages, but did not affect the host cells. NAIs therapy of L. major infected BALB/c mice resulted in reduced FP lesion size, diminished parasite burden, and importantly decreased induction of IL-4 and arginase activity in the presence of NAIs. In contrast IFN-γ and NO levels were significantly enhanced. NAIs therapy significantly diminished the progression of disease compared to the control group, but was less effective than amphotericin B treatment. CONCLUSIONS: Our study shows that NAIs treatment was effective in vitro and in Leishmania-infected mice, elicited a T-helper 1 (Th1) response and increased efficient cellular immunity, resulting in a diminished parasite load. Therefore, NAIs therapy can be considered as a useful and safe tool that can contribute to clearing L. major infections without inducing toxicity in host cells. The applications and mechanisms of NAIs therapy warrant further investigation especially in humans suffering from CL.

Arginase/nitric oxide modifications using live non-pathogenic Leishmania tarentolae as an effective delivery system inside the mammalian macrophages.

Recombinant live delivery system based on chemokine IFN-γ-inducible protein-10 kDa (CXCL 10 or IP-10), as a suitable immunotherapy tool, have been used for the treatment of Leishmania infections. This chemokine can defeat Leishmania spp. infection via producing nitric oxide (NO) for parasite killing. This study was performed to investigate the effects of IP-10 on the infected human macrophages by L. tarentolae expressing IP-10. We also quantified the arginase activity and NO production in the co-cultured human macrophages with L. tarentolae expressing IP-10 as compared with wild L. tarentolae. The results elucidate that in the infected cells with L. tarentolae expression of IP-10 the arginase activity decreased, and inversely, NO production intensely increased. Altogether, L. tarentolae expressing IP-10 shows a favorable therapeutic tool to improve the treatment of Leishmania infection. This work suggests that L. tarentolae expressing IP-10 cause specific effects on the metabolic pathways of the macrophage host, which might enable the host cells in killing of parasites and decreasing the survival of them against Leishmania infection.

Molecular Examination of Trichomonas vaginalis Infection and Risk of Prostate Cancer in the Biopsy of Patients with Different Prostate Lesions.

BACKGROUND: Trichomoniasis is a sexually transmitted infectious disease caused by a flagellated protozoa, Trichomonas vaginalis (T.vaginalis) and is often asymptomatic in men. Benign prostatic hyperplasia (BPH) and prostate cancer (PCA) are the most common urological diseases in the elderly. Scientists have proposed various factors which trigger prostate cancer, including sexually transmitted diseases. Thus, this study aimed to evaluate the potential role of T. vaginalis as a risk factor for various prostate lesions such as hyperplasia and prostate cancer. METHODS: A total of 250 paraffin-embedded of different prostate lesion biopsies were analyzed by Polymerase Chain Reaction (PCR) using the beta-tubulin gene for identifying T. vaginalis. RESULT: All 250 pathologic specimens were negative for this parasite by using PCR technique. CONCLUSION: It seems that T. vaginalis may have not had a causative role for different prostate lesions and it seems proposed PCR technique is an insufficient method to find the parasite in paraffin-embedded tissues. Therefore, other diagnostic techniques to identify the parasite in biopsy samples are suggested.

Anti-Leishmanial Activity of Artemisia persica, A. spicigera, and A. fragrance against Leishmania major.

BACKGROUND: Neglected tropical diseases (NTDs) like zoonotic cutaneous leishmaniasis (ZCL), is a widespread infectious disease with high mortality and morbidity. Various medications are used for treating the disease, but several side effects and drug resistance have been reported. Herbal medicines are unlimited sources for discovering new medications to treat infectious diseases. We aimed to determine the leishmanicidal activity of three species of Iranian Artemisia herbal plant extracts in in-vitro. METHODS: In-vitro anti-leishmanial activity of ethanolic extracts on both promastigotes and amastigotes was determined by using MTT method. IC50, CC50, EC50 and SI were calculated. The study was done in 2019-2020 in Iran University of Medical Sciences, Tehran, Iran. RESULTS: All of the three Artemisia species significantly reduced the number of parasite promastigotes. Among them, A. persica had the highest leishmanicidal activity against parasite promastigotes. Cytotoxicity assay elucidated that the Artemisia had no toxicity to the host cells, and killed the L. major amastigotes very efficiently. By increasing the dose of extracts, the parasite number in both phases (promastigotes and amastigotes) was reduced significantly. CONCLUSION: These results indicated satisfactory anti-leishmanial activity of Artemisia extracts against ZCL in-vitro. Accordingly, Artemisia ethanolic extracts might be considered as a strong, effective and safe herbal compound for clearing the L. major with less toxicity to the host macrophages cells. Hence, it may be recognized as an excellent herbal therapy for treating the ZCL.

Photodynamic Therapy Using Toluidine Blue O (TBO) Dye as a Photosensitizer against Leishmania major.

BACKGROUND: Photodynamic therapy (PDT) is alternative treatment of cutaneous leishmaniasis (CL), and phenolthiazine dyes such as Toluidine Blue O (TBO) have the potential role in PDT and notably affect parasites inactivation. This study aimed to evaluate the effectiveness of PDT by using TBO and a light-emitting diode (LED) in the treatment of zoonotic CL (ZCL). METHODS: The study was conducted in Iran University of Medical Sciences, Tehran, Iran in 2018-2020. Different concentration (7.8 µg/mL up to 500 µg/mL) of TBO as a photosensitizer and a 630 nm LED light as a source of light were used for antileishmanial activity against both forms of Leishmania major promastigotes and intracellular amastigotes. Effective concentration (EC50) and cell cytotoxicity (CC50) were calculated in both infected and non-infected J774.A1 macrophages, respectively. As well as inhibitory concentration (IC50) was quantified in L. major promastigotes for 2 h, 24 h, and 48 h after incubation using a MTT colorimetric assay. RESULTS: TBO dye in combination with the PDT significantly decreases the L. major promastigotes and intra-cellular amastigotes viability when compared with TBO alone. Both TBO dye in combination with the PDT and TBO alone had no toxic effects on the mice macrophages; however, it significantly killed the entered parasites inside the cells. Our results in the current study established satisfactory findings in clearing intracellular L. major parasites in in-vitro conditions. CONCLUSION: TBO dye in combination with the PDT can be considered as a harmless, effective and importantly perfect treatment against L. major, causative agent of ZCL, in an in-vitro situation without any negative toxicity to the mice macrophages.

Cryopreservation of Echinococcus granulosus Protoscoleces.

BACKGROUND: The main objective of the current study was to investigate on the cryopreservation of protoscoleces of Echinococcus granulosus, a causative agent of cystic hydatidosis in man. METHODS: This study was conducted on isolated protoscoleces from hydatid cysts infected livers collected from slaughterhouse of Tehran, Iran in 2016. Viability of protoscoleces was evaluated by dye test. Cryopreservation of isolated protoscoleces in the presence of Dimethylsulfoxide (DMSO) and glycerol using a three-step cooling protocol involving an initial period at -20 °C, -80 °C and liquid nitrogen was performed. RESULTS: The mean viability rate of 10% DMSO and 15% glycerol were 9% and 8% respectively. The protoscoleces of Echinococcus granulosus have been successfully thawed and recovered after 6 months storage in liquid nitrogen. CONCLUSION: Cryopreservation method needs to be improved for each species of helminthes and can be useful for other immunological and laboratorial studies.

How climate change can affect cholera incidence and prevalence? A systematic review.

Although the number of cholera infection decreased universally, climate change can potentially affect both incidence and prevalence rates of disease in endemic regions. There is considerable consistent evidence, explaining the associations between cholera and climatic variables. However, it is essentially required to compare and interpret these relationships globally. The aim of the present study was to carry out a systematic review in order to identify and appraise the literature concerning the relationship between nonanthropogenic climatic variabilities such as extreme weather- and ocean-related variables and cholera infection rates. The systematic literature review of studies was conducted by using determined search terms via four major electronic databases (PubMed, Web of Science, Embase, and Scopus) according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) approach. This search focused on published articles in English-language up to December 31, 2018. A total of 43 full-text studies that met our criteria have been identified and included in our analysis. The reviewed studies demonstrated that cholera incidence is highly attributed to climatic variables, especially rainfall, temperature, sea surface temperature (SST) and El Niño Southern Oscillation (ENSO). The association between cholera incidence and climatic variables has been investigated by a variety of data analysis methodologies, most commonly time series analysis, generalized linear model (GLM), regression analysis, and spatial/GIS. The results of this study assist the policy-makers who provide the efforts for planning and prevention actions in the face of changing global climatic variables.

Loop-Mediated Isothermal Amplification (LAMP) Assay to Detect Toxoplasmosis in Schizophrenia Patients.

BACKGROUND: Toxoplasma gondii (T. gondii) causes an important parasitic infection known as toxoplasmosis, which is a globally distributed important zoonosis. One of the major serious characteristics of T. gondii is its ability to manipulate the behavior of intermediate hosts. We performed a cross-sectional study to determine toxoplasmosis in schizophrenic patients, as one of the major neuropsychiatric disorders, using loop-mediated isothermal amplification (LAMP) technic by targeting parasite B1 gene. METHODS: Blood samples were taken from 118 schizophrenic patients hospitalized in tow hospitals including Baharan, Clinic of Psychiatric Ali-ibn-Abi-Talib Hospital (in Zahedan City), and Amir-al Momenin Psychiatric Hospital (in Zabol City), Sistan and Baluchestan Province, southeast Iran in 2016. They were analyzed using LAMP, and compared with the previous data of nested-PCR and serology. RESULTS: Out of the 118 schizophrenic individuals, 56 patients (47.4%) were found to be infected with T. gondii. The diagnosis of toxoplasmosis was confirmed in 41 patients (34.7%) via the nested-PCR. The seroprevalence of toxoplasmosis in schizophrenic patients was 55.9% (66/118). CONCLUSION: We found a high efficiency of LAMP method in identifying toxoplasmosis and its high prevalence among schizophrenic patients. Our findings could provide viable offer implications for the prevention of schizophrenia.

Antileishmanial activity of Urtica dioica extract against zoonotic cutaneous leishmaniasis.

BACKGROUND: Neglected parasitic diseases (NTDs) like cutaneous leishmaniasis (CL) have caused high mortality and morbidity rate in developing countries. This disease is considered as one of the six major tropical diseases, and has a great importance in HIV infected individuals as an opportunistic infection in those areas that both infections are endemic. This study evaluated the therapeutic effects of the Urtica dioica L (U. dioica) aqueous extract as an anti-leishmanial herbal drug in-vitro and in-vivo, and in addition to that, evaluated two vital immune system cytokines including gamma interferon (IFN-γ) and interleukin-4 (IL-4) plus nitric oxide (NO) and arginase activity against Leishmania major (L. major) infected mice. METHODOLOGY/PRINCIPAL FINDINGS: In-vitro anti-leishmanial activity of U. dioica aqueous extract was determined using MTT method and also Parasite Rescue Transformation Assay. Also, the footpad lesion size and parasite load in BALB/c mice infected with L. major were quantified for in-vivo assessment. Furthermore, for evaluating the immune responses, the levels of IFN-γ, IL-4, NO and arginase were measured in the BALB/c mice. These results indicated that U. dioica extract significantly reduced the L. major promastigotes viability. According to the in-vitro cytotoxicity assay of the extract on Leishmania parasites (CC50) and infected macrophages (EC50), the extract had no toxicity to the macrophages, however it efficiently killed the L. major amastigotes. In addition, the lesion size, parasite load, IL-4, and ARG were decreased in the treated infected mice, however IFN-γ and NO were significantly increased. CONCLUSIONS/SIGNIFICANCE: This study established satisfactory results in Leishmania parasite clearing both in-vivo and in-vitro. Therefore, U. dioica extract can be considered as an effective and harmless herbal compound for killing the parasite without toxicity to the host macrophages. Furthermore, it also can treat the CL by switching the mouse immune response towards a cell-mediated response (Th1); hence, it may be identified as a perfect therapeutic herbal drug for CL treatment.

Molecular Characterization of Fasciola spp. from Some Parts of Iran.

BACKGROUND: Identification of liver flukes, Fasciola hepatica, and Fasciola gigantica by morphometric parameters is not always reliable due to the overlapping measurements. This study aimed to characterize the liver flukes of animals from different parts of Iran by the genetic markers, ITS1, and COXI. METHODS: We collected flukes from infected livestock in six provinces of Iran from Sep to Nov 2016. The flukes were identified by amplification of a 680 bp sequence of ITS1 locus followed by a restriction fragment polymorphism (RFLP) assay. The genetic diversity among isolates was evaluated by amplification and sequencing of a 493 bp fragment of the COXI gene. RESULTS: We obtained 38 specimens from Khuzestan, 22 from Tehran, 10 from Isfahan, 10 from Mazandaran, 4 from Kurdistan, and 3 from Ardabil provinces. PCR-RFLP analysis revealed two patterns, representing F. hepatica, and F. gigantica. Fifty specimens from cattle and sheep exhibited F. hepatica pattern and 37 from the cattle, sheep, buffalo, and goat that of F. gigantica. The phylogeny based on COXI revealed two distinct clades separating F. hepatica from F. gigantica. In our phylogeny, the Iranian F. gigantica isolates showed a distinct separation from the African flukes, while grouped with the East Asia specimens demonstrating a common ancestor. The F. hepatica isolates clustered with the flukes from different parts of the world, including East Asia, Europe, and South America. CONCLUSION: The present study revealed a substantial genetic difference between F. gigantica populations of Asia and Africa, while F. hepatica isolates from different parts of the world shared high similarities.