The Effect of Curcumin on the Expression of INFγ, TNF-α, and iNOS Genes in PBMCs Infected with Leishmania major [MRHO/IR/75/ER].

BACKGROUND: Leishmaniasis, caused by the Leishmania parasite, is one of the most important tropical neglected diseases. The urgent search for effective, inexpensive, and preferably herbal anti-leishmanial agents, is needed. OBJECTIVE: Curcumin is a natural polyphenolic compound derived from turmeric that is well known for its antioxidant, anti-inflammatory, anti-tumor, and anti-cancer activity. METHODS: The present work evaluates the anti-leishmanial [Leishmania major] activity of curcumin. The infected PBMCs were treated with curcumin. The ROS level at 6, 12, 24 h and gene expression levels at 24, 48, and 72 h of PBMCs after treatment with curcumin were determined. RESULTS: Based on the results, the curcumin concentrations of 268 µM [24 h] and 181.2 µM [72 h] were defined as IC50 against L. major promastigotes. Treatment of L. major infected-peripheral blood mononuclear cells [PBMCs] with IC50 concentrations of curcumin, depending on exposure time, significantly induced the reactive oxygen species [ROS] generation and increased the expression levels of interferongamma [IFN-γ], tumor necrosis factor-alpha [TNF-α], and nitric oxide synthase [iNOS] genes. CONCLUSION: These findings suggest the potential of curcumin against Leishmaniasis.

Anti-toxoplasma and Cytotoxic Activities of Holothuria leucospilota Extract and TiO2NPs In vitro and In vivo.

BACKGROUND: An impressive treatment for toxoplasmosis is the combinatory use of sulfadiazine and pyrimethamine. However, both the drugs involve significant side effects and toxicity for the host. Therefore, the discovery of new anti-toxoplasma medications with high efficacy and less to no side effects is urgently needed. OBJECTIVE: This study aimed to evaluate the anti-toxoplasmic effects of Holothuria leucospilota (H. leucospilota) extract and TiO2NPs on the cell death of Toxoplasma gondii (T. gondii) tachyzoites in vitro and serum liver enzymes (AST, ALT, and ALP), and also to evaluate the immune response and production of IL-5, IFN-γ, and TNF-α in a mouse model. MATERIALS AND METHODS: The cytotoxicity of TiO2NPs and H. leucospilota extract against the tachyzoite of T. gondii was evaluated by the methyl thiazolyl tetrazolium (MTT) assay. The levels of serum TNF-α, IFN-γ, IL-5, and liver enzymes were measured, as well. All the groups were subjected to T. gondii, and the survival rate of experimental mice was evaluated. RESULTS: Our findings suggested in vivo and in vitro anti-toxoplasmic activity of TiO2NPs and H. leucospilota extract by inhibiting the proliferation and invasion of T. gondii tachyzoite. In addition, a significant increase in IFN-γ and TNF-α production was observed in mice treated with high doses of TiO2NPs and H. leucospilota extract. However, IL-5 levels decreased in TiO2NPs and H. leucospilota extract-treated mice. Our results also showed a highly significant increase (P < 0.05) in the levels of ALT, AST, and ALP in the groups injected with TiO2NPs and H. leucospilota extract, but not the control group. CONCLUSION: TiO2NPs and H. leucospilota extract have greater anti-toxoplasma effects in vitro and in vivo. These two compounds could be considered as a candidate for use against toxoplasmosis, both therapeutically and prophylactically.

Screening of Cystic Echinococcosis and Toxocariasis in Urmia Municipal Workers, Northwest Iran.

BACKGROUND: Echinococcus granulosus and Toxocara spp., the causative agents of hydatid cyst and toxocariasis in humans and animals, are zoonotic infections with medical and veterinary significance respectively. AIM: Herein, we aimed to investigate the seroprevalence of toxocariasis and cystic echinococcosis in Urmia municipal workers (gardeners and sweepers) in 2018 using ELISA assay. OBJECTIVE: There is no precise report on the seroprevalence of toxocariasis and cystic echinococcosis in Urmia municipal workers using ELISA assay. Therefore, this study found an opportunity to investigate this subject. METHODS: A total of 220 blood samples were obtained from municipal workers with an average age of 41.31 ± 9.42. Commercial anti-Toxocara canis and hydatid cyst ELISA kits were used to assess the seroprevalence rates. RESULTS: Approximately, 14.5% and 2.3% of individuals were seropositive for toxocariasis and hydatidosis, respectively. Consuming unboiled milk and/or raw/uncooked meat and rural conditions are risk factors for toxocariasis. Regarding hydatidosis, consuming unwashed vegetables was the only statistically significant risk factor. Toxocara infection (p = 0.01), but not hydatid cyst (p = 0.05), is more prevalent among sweepers than municipal gardeners. CONCLUSION: Due to the occupational condition of municipal workers, they are exposed to many infectious agents; thus, more researches should be done on these populations.

Prevalence of IgG and IgM anti-Toxoplasma gondii Antibodies in Blood Donors at Urmia Blood Transfusion Organization, Iran.

OBJECTIVE: The importance of toxoplasmosis lies in its global spread, opportunistic nature, and causative role in abortion or irreparable adverse effects on infants of infected pregnant women. Toxoplasma gondii has different transmission routes to humans, including blood transfusion. The objective of the present study was to evaluate the prevalence of IgG and IgM antibodies specific for T. gondii in blood donors at the Urmia Blood Transfusion Organization in west Azerbaijan, Iran. METHODS: The present analytical, descriptive study evaluated the plasma of 270 randomly selected blood bags donated in 2013. The enzyme-linked immunosorbent assay (ELISA) test was employed to measure anti-T. gondii IgG and IgM antibodies. RESULTS: The results of the ELISA test showed that 102 samples (37.8%) from 270 blood bags had IgG antibodies in their plasma and none of them were IgM-positive, whereas 98 were men and four were women. CONCLUSION: Any increase in the level of IgM antibodies indicates the presence of an acute disease because the parasite is inside white blood cells and contaminates blood transfusion. Fortunately, all samples were IgM-negative. However, a province-wide seroepidemiological study is required for the Blood Transfusion Organization to consider including screening for anti-T. gondii antibodies in its screening programs.

Antileishmanial and Immunomodulatory Activity of Allium sativum (Garlic): A Review.

Leishmaniasis is caused by an obligate intracellular protozoa belonging to Leishmania genus. The current drugs for treatment of leishmaniasis possess many disadvantages; therefore, researchers are continuously looking for the more effective and safer drugs. The aim of this study is to review the effectiveness, toxicities, and possible mechanisms of pharmaceutical actions of different garlic extracts and organosulfur compounds isolated from garlic against Leishmania spp. in a variety of in vitro, in vivo and clinical trials reports. All relevant databases were searched using the terms "Allium sativum," "Garlic," "Allicin," "Ajoene," "Leishmania," "in vitro," "in vivo," and "clinical trial," alone or in combination from 5 English databases (Web of Science, PubMed, Science Direct, Scopus, Google Scholar) and 3 Persian databases (Scientific Information Database, Iran Medex, and Magiran) from 1990 to 2014. In summary, garlic with immunomodulatory effects and apoptosis induction contributes to the treatment of leishmaniasis.

Evaluation of the alum-naloxone adjuvant activity against experimental murine leishmaniasis due to L. major.

Leishmaniasis is caused by intracellular parasites of Leishmania species, which are transmitted by the bite of the sandfly. Recovery and protection against the infection depends on the induction of a strong Th1 type of immune response. Vaccination of mice with the opioid antagonist naloxone can promote the activation of the Th1 responses. We studied the efficacy of the mixture of naloxone and alum, as an adjuvant, to enhance immune responses and induce protection against Leishmania major infection in BALB/c as a susceptible mouse model. BALB/c mice were immunized with Ag-naloxone-alum, Ag-alum, Ag-naloxone or PBS subcutaneously three times at 2-week intervals. The humoral and cellular specific immune responses were assessed 2 weeks after the last immunization and compared with the control mice. Our results indicated that the administration of alum-naloxone as an adjuvant increased the capability of L. major promastigote antigens to enhance lymphocyte proliferation, the levels of IFN-γ, and the IFN-γ/IL-5 ratio. The results of DTH showed that there were no significant differences in footpad swelling between the groups of immunized mice as compared with the non-vaccinated control group; however, no significant differences were observed in the survival rate among groups. It can be concluded that although immunization with the alum-naloxone mixture in combination with the autoclaved L. major promastigote antigens could enhance cellular immunity and shift the immune response to a Th1 pattern, it could not protect the mice against Leishmania major infection.

A novel adjuvant, the mixture of alum and naltrexone, augments vaccine-induced immunity against Plasmodium berghei.

We previously showed that the mixture of naltrexone (NLT), a general opioid antagonist, and alum, acts as an effective adjuvant in enhancing vaccine-induced T helper 1 (TH1) humoral immune responses against Toxoplasma gondii. Here, we tested the efficacy of the mixture of NLT and alum in the induction of immunity in response to blood stages of Plasmodium berghei (BSPb) as a model vaccine. BALB/c mice were divided into five vaccination groups. Mice in the experimental groups received the BSPb vaccine alone or in combination with the adjuvant alum, NLT or the alum-NLT mixture. Mice in the control group received PBS. All mice were immunized on days 0, 7 and 14. Two weeks after the last immunization, immune responses to Plasmodium berghei were assessed. Our results indicated that including the alum-NLT mixture as an adjuvant during vaccination increased the ability of the BSPb vaccine to enhance lymphocyte proliferation, shifted the immune response towards a TH1 profile and increased Plasmodium berghei-specific IgG2a. This resulted in improved protective immunity against Plasmodium berghei. In conclusion, administering alum-NLT mixture in combination with the BSPb vaccine enhanced the vaccine-induced immunity, and shifted the immune response toward TH1 pattern.

A novel adjuvant, mixture of Alum and naltrexone, elicits humoral immune responses for excreted/secreted antigens of Toxoplasma gondii tachyzoites vaccine in Balb/c murine model.

OBJECTIVE: The excreted-secreted antigens (ESA) from the tachyzoites seem to play a key role in immunity against Toxoplasma gondii. The aim of this study is to investigate whether Alum-NLT mixture, as a new adjuvant, can induce humoral immunity in response to excreted secreted antigens (ESA) of Toxoplasma gondii as a model vaccine or not. METHODS: Six- to eight-week-old female Balb/c mice were divided into five groups. Mice in the experimental groups received either ESA vaccine alone or in combination with the adjuvant Alum, NLT or Alum-NLT mixture; Mice in the negative control group received phosphate buffered saline (PBS). All mice were immunised, three times subcutaneously (s.c.) with a total volume of 150µl each with a 10-day interval. Ten days after the final immunisation, immune response to Toxoplasma gondii was assessed. RESULTS: Our results revealed that Alum-NLT mixture as an adjuvant during vaccination boosts the efficacy of the ESA vaccine by means of increasing Toxoplasma gondii-specific IgG, IgG2a production and the ratio of IgG2a/IgG1 (P-value < 0.05). The use of this adjuvant mixture improved the protective immunity against Toxoplasma gondii. CONCLUSION: Administration of the Alum-NLT mixture as an adjuvant in ESA vaccine enhances humoral immunity.

Identification of Echinococcus granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan - Iran.

BACKGROUND: The aim of this study was DNA extraction from protoscolecses of Echinococcus granulosus and identification of these strains in West-Azerbaijan Province, north western Iran. METHODS: Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different restriction enzymes of Taq 1, Rsa 1 and Alu 1. RESULT: Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex). The results of RFLP analysis also were the same for all isolates. PCR-RFLP patterns restriction enzymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approximately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples. CONCLUSIONS: Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex) is dominant genotype in this province.

Comparison of four methods for DNA extraction from Echinococcus granulosus protoscoleces.

OBJECTIVE: Cystic echinococcosis caused by the dog tapeworm Echinococcus granulosus, is a global zoonotic infection which is economically important and constitutes a major threat to public health in many countries, especially in Middle Eastern countries. Strain characterization is essential for the establishment of a preventive and control strategy in every endemic area. Before all molecular studies, it is necessary to achieve DNA of the parasite. The aim of this study was to compare four simple methods of DNA extraction from protoscolecses of E. granulosus in samples from the West Azerbaijan province of Iran. METHODS: After collecting sheep and cow hydatid cysts from several slaughterhouses of the province, DNA samples were extracted using four different methods involving the use of glass beads, commercial DNA extraction kit, boiling and crushing. For all DNA samples extracted, electrophoresis on 1.3% agarose gel was performed. RESULTS: The methods were compared regarding DNA, time and cost effectiveness and facility. All methods showed some advantages and disadvantages. The commercial kit method, which was the most time and cost effectiveness method, achieved no bands and glass beads method had the best results on gel electrophoresis. CONCLUSION: Glass beads and boiling methods were the most suitable methods regarding their ease, quickness and cost effectiveness.

Role of iron supplementation in promoting maternal and fetal outcome.

BACKGROUND: The data comparing daily versus intermittent iron supplementation during pregnancy remain controversial. This study was undertaken to compare the efficacy of daily versus two different intermittent iron supplementation regimes on hematologic markers and birth outcomes in nonanemic pregnant women. METHODS: Two hundred and ten women with singleton pregnancies, no known disease, and hemoglobin levels >11.0 g/dL were randomly assigned to one of three groups, ie, Group A consuming two iron supplementation tablets once weekly (100 mg iron per week, n = 70), Group B consuming one tablet twice weekly (100 mg iron per week, n = 70) and Group C, consuming one tablet daily (50 mg iron per day, n = 70). No additional micronutrients were supplied. Hemoglobin and serum ferritin levels were measured at 20, 28, and 38 weeks. Pregnancy and birth outcomes (pregnancy termination, method of delivery, birth weight, stillbirth) were analyzed. RESULTS: In total, 201 women completed the protocol. There was a significant difference in mean hemoglobin and ferritin levels in Group B at 38 weeks (P = 0.018 and P = 0.035, respectively) but this difference was not clinically significant (hemoglobin >12 g/dL, ferritin >19 µg/L). There was a significant increase in ferritin in Group C (P = 0.03) at 28 weeks. No significant difference was observed with respect to pregnancy or birth outcome across the groups. All regimens prevented the occurrence of hemoglobin <10.5 g/dL, but weekly supplementation was associated with development of a hemoglobin level <11.0 g/dL (risk ratio 0.044). CONCLUSION: Twice-weekly supplementation is as effective as daily supplementation, and may represent an acceptable compromise in iron supplementation regimens for nonanemic pregnant women.

Production of experimental hydatid cyst in the eye, peritoneum and liver of BALB/c mice.

As the main treatment for this infection is surgery, the surgery team personnel are at the risk of the protoscoleces released from the hydatid cysts (HC) of patients. The first goal of this study was to determine the probability of the production of ocular HC in mice due to the fluid of the aspirated protoscoleces from the sheep liver with HC. The second goal of this study was to produce HC in the peritoneum and liver, in order to gather more information for future studies on hepatic and peritoneal HC treatment procedures. For the first goal of this study, different concentrations of protoscoleces were prepared and injected into the eyes of 60 mice. After 20 weeks, 10 of the 60 mice of this group died. The remaining 50 mice were examined by a surgeon under the anesthesia. There weren't any symptoms of HC in the eyes and around it. For the second goal, 39 new mice were separated into three sub groups and 0.5 ml of protoscolex solution was injected intraperitoneally. After 20 weeks, they were anesthetized and their peritoneum, intestines and liver were examined. HC was seen in the peritoneum and liver of 6 mice.