Hepatoprotective effect of Descurainia sophia seed extract against paracetamol-induced oxidative stress and hepatic damage in mice

Introduction: In this study, we aimed to evaluate the protective effects of Descurainia sophia seed extract on paracetamol-induced oxidative stress and acute liver injury in mice. Methods: Sixty male Swiss albino mice were divided into 6 groups, group I: as control group received NaCl 9%. Group II: as paracetamol group received paracetamol intraperitoneally (i.p.) (500 mg/kg). Groups III to VI as treatment groups received different doses of D. sophia seed extract (T1 = 50, T2 = 100, T3 = 200 and T4 = 300 mg/kg, respectively). Twenty-four hours after the paracetamol administration, the mice were sacrificed under mild anesthesia and their blood samples were collected to estimate serum alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin and malondialdehyde (MDA) levels. Their livers were also removed for histopathological examinations. Results: Pretreatment of mice with D. sophia seed extract, significantly prevented the paracetamol induced elevation in the levels of serum ALT, AST and ALP, total bilirubin and MDA (P < 0.05). The results of histopathologic studies were consistent with the above findings. Conclusion: Descurainia sophia seed extract has a protective effect against paracetamol induced oxidative damage and acute hepatotoxicity in mice

Molecular detection of Brucella melitensis in sheep and goat milk in Iran

Purpose: To detect Brucella melitensis in the milk of reared sheep and goats from Isfahan and Shahrekord regions, Iran.Methods: A total of 225 milk samples (sheep = 125; goat = 100) were collected from Isfahan and Shahrekord regions, Central Iran. Polymerase chain reaction (PCR) was used to detect the presence of B. melitensis in the milk following standard procedures.Results: From 225 milk samples, 20 (8.9 %) were positive for B. melitensis. Out of 125 sheep milk, 12 (9.6 %) had B. melitensis, and of these, 8 (66.6 %) were milk collected from Shahrekord and 4 (33.3 %) from Isfahan region. On the other hand, out of 100 goat milk samples, 18 (18 %) were positive for B. melitensis, out of which 10 (55.5 %) were from Shahrekord and 8 (44.4 %) from Isfahan.Conclusion: The findings show that B. melitensis is present in a significant proportion of caprine and ovine milk in a section of Iran.Keywords: Brucella melitensis, Milk, Polymerase chain reaction, Sheep, Goa

Unresponsiveness to Glucantime Treatment in Iranian Cutaneous Leishmaniasis due to Drug-Resistant Leishmania tropica Parasites

BACKGROUND: Recent circumstantial evidence suggests that an increasing number of Iranian patients with cutaneous leishmaniasis are unresponsive to meglumine antimoniate (Glucantime), the first line of treatment in Iran. This study was designed to determine whether the clinical responses (healing, or non-healing) were correlated with the susceptibility of Leishmania parasites to Glucantime. METHODS AND FINDINGS: In vitro susceptibility testing was first performed on 185 isolated parasites in the intracellular mouse peritoneal macrophage model. A strong correlation between the clinical outcome and the in vitro effective concentration 50% (EC (50)) values was observed. Parasites derived from patients with non-healing lesions had EC (50) values at least 4-fold higher than parasites derived from lesions of healing patients. A selection of these strains was typed at the molecular level by pulsed-field gels and by sequencing the pteridine reductase 1 (PTR1) gene. These techniques indicated that 28 out of 31 selected strains were Leishmania tropica and that three were Leishmania major. The L. major isolates were part of a distinct pulsed-field group, and the L. tropica isolates could be classified in three related additional pulsed-field groups. For each pulsed-field karyotype, we selected sensitive and resistant parasites in which we transfected the firefly luciferase marker to assess further the in vitro susceptibility of field isolates in the monocyte cell line THP1. These determinations confirmed unequivocally that patients with non-healing lesions were infected with L. tropica parasites resistant to Glucantime. Additional characterization of the resistant isolates showed that resistance is stable and can be reversed by buthionine sulfoximine, an inhibitor of glutathione biosynthesis. CONCLUSIONS: To the authors' knowledge, this is the first report of proven resistant parasites contributing to treatment failure for cutaneous leishmaniasis and shows that primary Glucantime-resistant L. tropica field isolates are now frequent in Iran

Evaluation of Immune Response against Leishmaniasis in BALB/c Mice Immunized with Cationic DOTAP/DOPE/CHOL Liposomes Containing Soluble Leishmania major Antigens

Background: Whole killed Leishmania vaccine reached phase III clinical trials but failed to display significant efficacy in human mainly due to limited Th1 inducer adjuvant. Liposomes consisting of 1, 2-dioleoyl-3trimethylammonium-propane (DOTAP) bearing an inherent adjuvanticity and 1, 2-dioleoyl-L-α-glycero-3-phosphatidylethanolamine (DOPE) is well known to intensify the efficacy of positively charged liposomes. Methods: Soluble Leishmania major antigens (SLA) encapsulated in cationic liposomes using lipid film method in 2016). BALB/c mice were immunized subcutaneously (SC), three times in a 2-wk interval, with Lip (DOTAP)-SLA+, Lip (DOTAP/DOPE)-SLA+, Lip (DOTAP/DOPE/CHO)-SLA+, Lip (DOTAP/DOPE/CHO), Lip (DOPE/CHO), SLA or HEPES buffer. At week 2 after the last booster injection, immunized mice have challenged SC in the footpad with L. major parasites. To investigate the rate of protection and the type of immune response generated in mice, lesions development was assessed, IL-4 and IFN-γ levels with the ratio of IgG2a/IgG1 isotype were studied to describe the type of generated immune response. Results: Mice immunized with all liposomal form of SLA showed smaller footpad swelling and lower parasite burden in the spleen and footpad compared to the group of mice received buffer. However, these formulations did not show protection against leishmaniosis because of a generated mixed Th1/Th2 response in mice characterized by high production of IFN-γ and IL4 and a high titer of IgG1 and IgG2a antibody. Conclusion: Immunization with Lip (DOTAP/DOPE/CHO)-SLA+ was not an appropriate strategy to protect mice against leishmaniosis

The changing profile of cutaneous leishmaniasis agent in a central province of Iran

Cutaneous leishmaniasis in Iran is usually caused by Leishmania major or L. tropica. However, the direct examination or the cultures of biopsies for diagnosis are not very sensitive. The objective of this study was to identify the responsible species obtained from patients suspected of cutaneous leishmaniasis referred to the reference laboratory at Yazd in Iran during 2010-2011 using parasitological and molecular assays. After completing a clinical/epidemiologic data questionnaire for 145 patients with suspected skin lesions, scraping samples were collected. Each specimen was examined using both direct microscopy and molecular assay using polymerase chain reaction-restriction length polymorphism (PCR-RFLP). Location of the lesions included 47.7% on hands, 30.7% on face, 15.4% on feet, and the remainder on other regions. Out of 145 samples, Leishman body was observed in 52 by direct smear and 73 by PCR assay. Molecular assay indicated 36 cases as L. major, 36 cases as L. tropica and one case as unknown.  In conclusion, molecular characterization showed changing profile of Leishmania species in the study area which may have implications on treatment and/or control strategies

Iranian Native Plants on Treatment of Cutaneous Leishmaniosis: A Narrative Review

Background: Chemotherapy still relies on the use of pentavalent antimonials, amphotericin B, paromomycin, miltefosin, and allopurinol. In this study, we explained about the native plant that grows in different regions of Iran and used as anti-leishmanial in Iran and even many other countries. Methods: This narrative review covers all information about local herbal medicine in Iran that used in treatment of cutaneous leishmaniasis in all the worlds, published in local and international journals from 1996 to 2015 using various databases including PubMed, SID, Google Scholar, Scopus, and Science Direct. Results: Overall, 150 articles in databases were identified. Many local plants grown in some places of Iran were used to treat this endemic disease. Conclusion: The cutaneous leishmaniasis is also a major health problem in Iran, especially in Mashhad (Northeast of Iran). Therefore, many patients seek for herbal therapy that is cheaper and readily available. This review provides information regarding plant that exists in Iran and exhibiting effects on anti- Leishmania activity. Among the anti-leishmanial mentioned in this review, most have never been tested for cytotoxicity and very few have been tested for in vivo activity

Evaluation of a Possible Synergistic Effect of Meglumine Antimoniate with Paromomycin, Miltefosine or Allopurinol on in Vitro Susceptibility of Leishmania tropica Resistant Isolate

Background: Pentavalent antimonials are still the first choice treatment for leishmaniasis, but with low efficacy and resistance is emerging. In the present study, the effect of meglumine antimoniate (MA, Glucantime) combined with paromomycin, miltefosine or allopurinol on in vitro susceptibility of Leishmania tropica resistant isolate was evaluated. Method: The drugs were obtained from commercial sources and diluents of each drug in medium were prepared on the day of experiment. J774 A.1 murine macrophage cell lines were attached to the cultured on slide and incubated at 37 0C with 5% CO2 for 24 h. Then the stationary phase promastigotes were added to the cells and after 4 hrs of incubation different concentrations of MA, paromomycin, miltefosine or allopurinol were added and incubated for an additional of 72 h. Then the slides were dried and fixed with methanol, stained by Giemsa and studied under a light microscope. Drug activity was evaluated by assessing the macrophage infection rate and the number of amastigotes per infected macrophage was done by examining 100 macrophages. The experiment was done in triplicates. Result: Various concentrations of MA along with paromomycin, miltefosine or allopurinol significantly inhibited (P<0.01) the proliferation of L. tropica amastigote stage in the macrophage cell line as compared with MA alone or positive control. Conclusion: Combination of Glucantime with paromomycin, miltefosine or allopurinol showed a synergistic effect on the clinical isolate of L. tropica in vitro. Use of combination therapy is a new hope and a logical basis for therapy of the patients with cutaneous leishmaniasis. Further investigations are needed to evaluate the therapeutic effects of these drugs on the CL patients

Modulation of the Immune Response to DNA Vaccine Encoding Gene of 8-kDa Subunit of Echinococcus granulosus Antigen B Using Murine Interleukin-12 Plasmid in BALB/c Mice

Abstract Background: The current study was designed to evaluate immune responses induced by DNA vaccines encoding 8-kDa subunit of antigen B (HydI) of Echinococcus granulosus and murine interleukin 12 (IL-12) as genetic adjuvants in BALB/c mice. Methods: Expression plasmid pcDNA3.1 containing HydI (pcHyd1) as vaccine along with the murine interleukin 12 (pcMIL12) as adjuvant were used. Thirty-five mice in the five experimental groups received PBS, empty pcDNA3.1, pcHydІ, pcMIL-12, and pcHydІ+ pcMIL-12 in days zero, 14th and 28th. Two weeks after the last immunization, evaluation of the immune response was performed by evaluating the proliferation of splenic lymphocytes, IFN-γ and IL-4, determination of IgG isotyping titer. Results: Mice that received the pcHydI+pcMIL12 exhibited higher levels of lymphocyte proliferation compared to mice that received the pcHydI alone (P<0.001), and produced significantly more IFN-γ in comparison to other groups (P< 0.001). In addition, they produced significantly less IL-4 than mice receiving the PBS and the empty plasmid (P<0.023). The IgG2a levels were clearly higher in pcHydI+pcMIL12 group in comparison with the groups of pcHydI alone, empty plasmid, and PBS. In contrast, IgG1 was elevated in the group of pcHydI. Conclusion: Co-delivery of IL-12 with DNA encoding 8-kDa subunit of antigen B was effective significantly in inducing the immune response in mice

A Prospective Cohort Study of Cutaneous Leishmaniasis Risk and Opium Addiction in South Eastern Iran

Opium addiction and cutaneous leishmaniasis (CL) are endemic in different parts of Iran, particularly in Bam, where a massive earthquake occurred. This study was designed to compare the incidence rate and severity of CL cases among opium addicted and non-addicted individuals in south-eastern Iran. This study was carried out as a prospective cohort by active house-to-house visits of 1,481 habitants in Bam. CL cases were confirmed by smear and identification of Leishmania species was performed using nested-PCR. The data was analyzed by x2 and t-tests, using SPSS software and also KaplanMeier survival curve and long-rank test in Stata 11.2 and P,0.05 was considered as significant. A total of 904 individuals consisting of 226 opium addicted and 678 non-addicted individuals were followed-up for a period of seven years. The two cohorts were similar in terms of age, sex and place of residency. A similar pattern of incidence was observed among the two cohort groups. In contrast, the severity of CL in terms of the number, duration and the size of the lesions in opium addicted individuals was significantly (P,0.001) higher than non-opium addicted individuals. In conclusion, the present findings indicate that there is no relationship between the incidence of CL and opium addiction

Emergence of a new focus of anthroponotic cutaneous leishmaniasis due to Leishmania tropica in rural communities of Bam district after the earthquake, Iran

objectives To describe a new emerging focus of anthroponotic cutaneous leishmaniasis (ACL) due to Leishmania tropica in rural areas of Dehbakry county, south-eastern Iran, after the earthquake of 2003. methods House-to-house survey of 3884 inhabitants for active leishmaniasis lesions or scars. The diagnosis was confirmed by smears, cultures and identification of the parasite by polymerase chain reaction (PCR). results All age groups were affected, although patients £10 years of age showed the highest rate of infection (P = 0.0001). The overall prevalence rate was 5.3%; 6.3% in females and 4.3% in males. Of 204 cases, 1.8% had active sores and 3.5% had scars, with a significant difference between the sexes (P = 0.005). 47% of the lesions were on the face and 77.9% had one lesion. The incidence rose gradually 2004–2005, but grew exponentially 2006–2008. Electrophoresis of PCR products indicated that L. tropica was the causative agent. conclusions The current emergence was unexpected in this rural locality, where no previous history of CL was recorded. According to our knowledge this is the first report of a gradually establishing new ACL focus in rural communities after the 2003 earthquake