Potential Protective Role of Cyrtopodion Scabrum in Antioxidant Parameters in Serum and Liver of Rats with 5-FU-Induced Oxidative Damage.

Chemotherapy is the main approach for the treatment of cancer; however, it often causes unpleasant oxidative damages. Therefore, the development of an effective alternative/complementary therapy with improved tumor suppression efficiency and lower adverse effects is highly required. Recently, it has been shown that Cyrtopodion scabrum extract (CsE) is an effective and selective tumor suppressor medicine. The present study investigated the antioxidant activity of Cyrtopodion scabrum homogenate (CsH) and CsE and their effects on attenuating 5-fluorouracil (5-FU)-induced liver dysfunction in rats. A total of 60 male rats (weight: 200-220 g) were divided into six groups and treated for 14 days. The control (group I) and 5-FU (group II) groups received distilled water and 5-FU, respectively. The other four groups were orally administered with CsE, CsH, CsE+5-FU, and CsH+5-FU (groups III to VI), respectively by gavages based on a daily schedule. The 5-FU-induced oxidative damage was evaluated by changes in the weight and food and water intake during the treatment and antioxidant parameters in the liver and serum of the treated rats. The obtained data indicated that the administration of CsH and CsE significantly improved liver function and defense system of antioxidants by attenuating the levels or activities of malondialdehyde, superoxide anion, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase and decrease of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, total antioxidant capacity, glutathione, total protein, and albumin in the liver and serum, induced by 5-FU treatment. The obtained data of the current study suggested that CsH and CsE play a protective role in the imbalance elicited by 5-FU and can be used as alternative/complementary supplements with 5-FU to reduce oxidative damages which is the consequence of reactive oxygen species production in cancerous patients.

Clinical and laboratory evaluation of cured and non-cured patients with cutaneous leishmaniasis treated by Glucantime.

BACKGROUND & OBJECTIVES: Insufficient treatment of cutaneous leishmaniasis (CL) by conventional drugs is a major barrier in control strategies. This study was aimed to evaluate Glucantime efficacy and the susceptibility of Glucantime unresponsive and responsive CL isolates in the field and laboratory. METHODS: Chi-square test (x[2]) was used to determine the significance of difference between proportions in Glucantime-treated patients. The inhibitory activity of various concentrations of Glucantime against Leishmenia tropica stages was evaluated by a colorimetric cell viability MTT and macrophage assays. Mixed model, t-test and ANOVA were performed to determine the significance of difference between various concentrations of Glucantime unresponsive or responsive isolates and untreated control group and p <0.05 was defined as significant level. Altogether, 89.8% of the patients were cured by Glucantime, whilst 10.2% remained non-cured. RESULTS: The overall Glucantime efficacy in different age groups and genders was similar. The IC50 values of promastigotes and amastigotes for Glucanime unresponsive isolates were 2.1 and 2.6 times higher than the equivalent rates obtained for responsive cases, respectively. The overall mean number of amastigotes within macrophages in unresponsive isolates was significantly higher (32.68 ± 1.24) than that in responsive ones (18.68 ± 1.52, p <0.001). Glucantime unresponsive and responsive field isolates of anthroponotic CL (ACL) caused by L. tropica strongly correlated to in vitro assays. INTERPRETATION & CONCLUSION: Monitoring of Glucantime unresponsiveness by the health surveillance system is extremely important, where anthroponotic transmission occurs in humans. Hence, physicians should be aware of such clinical unresponsive presentations with ACL for antimonial therapeutic failure to improve management of disease in endemic regions.

A newly emerged focus of zoonotic cutaneous leishmaniasis in South-western Iran.

Leishmaniasis is rising in many countries, including Iran, due to climate change, refugee crises, urbanization and etc. The aim of this study was to explore the epidemiology, extent and identity of Leishmania species in a newly emerged focus in Abdanan County, Ilam Province, South-western Iran. This study was performed as a descriptive cross-sectional study by a systematic house-to-house approach. The Leishmania species was identified by RFLP-PCR and sequencing. Altogether, 46799 individuals consisting of 22907 (48.9) female and 23892 (51.1%) male were interviewed and physically examined for the presence of skin lesions. Overall, the incidence rate was 0.34% (n = 160). All age groups were affected and the incidence rate was the highest in <10 years of age group (0.49%) and the lowest in >50 years old individuals (0.15%), although there was no significant difference regarding the sex and age. The majority of patients had one lesion (47.5%) on hands (56%) and most of the cases occurred in Abdanan city (%54) in summer. Based on the RFLP-PCR analysis, all the Leishmania isolates were L. major of single genotype. A newly emerged focus of zoonotic CL caused by L. major occurred in South-western of Iran. Multiple risk factors created this epidemic area. Further studies on the vector and reservoir are crucial needed to provide evidences to select the prophylactic and therapeutic measures for future control strategies.

Molecular detection and genetic diversity of Toxoplasma gondii in different tissues of sheep and goat in Eastern Iran.

This study was designed to detect parasitic DNA in tissues from sheep and goats raised and slaughtered in the southeastern Iran as well as to genetically characterize infecting strains of T. gondii. A total of 240 tissue samples consisting of heart, brain, and diaphragm were obtained from sheep (n=40) and goats (n=40) slaughtered in abattoirs from three provinces located in southeastern Iran including Kerman, Razavi Khorasan, and South Khorasan Provinces between February to October 2015. Nested PCR amplified the B1 and GRA6 genes. To determine the genetic characterization of positive samples, all genotyped positive samples were examined by PCR-RFLP. Sequencing analysis was performed to evaluate the prevalence of type strains (I, II and III). A total of 68(56.66%) tissue samples of sheep and 53(44.16%) from goats were found to be positive for T. gondii B1 gene, that included 11(27.5%) diaphragm, 21(52.5%) heart, and 36(90%) brain of sheep; and 20(50%) diaphragm, 11(22%) heart and 22(55%) brain of goats. Moreover, 22(18.3%) tissue samples of sheep and 20(16.6%) tissue samples of goats were found positive with GRA6 gene for T. gondii. There are three genotypes and mix genotype using mseI enzyme among all positive samples. The results demonstrated the presence of T. gondii DNA in tissues of sheep and goats from southeast of Iran. Control of Toxoplasma infection animal products are important in consumer protection.

Fabrication and characterization of non-enzymatic glucose sensor based on ternary NiO/CuO/polyaniline nanocomposite.

Novel nickel and copper oxide nanoparticle modified polyaniline (PANI) nanofibers (NiO/CuO/PANI) were fabricated and used as a non-enzymatic sensor for detecting glucose. PANI nanofibers were prepared through electrodeposition, whereas nickel and copper oxide nanoparticles were deposited on PANI nanofibers by electrodeposition and electrochemical oxidation in situ. The morphology and structure of NiO/CuO/PANI nanocomposites were characterized by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Raman spectroscopy, and Fourier transform infrared (FT-IR). The as-prepared NiO/CuO/PANI electrode was employed for non-enzymatic glucose detection in alkaline electrolyte and showed better electrocatalytic activity compared with the PANI, CuO/PANI, and NiO/PANI electrodes. Consequently, an amperometric electrode of glucose was achieved under 0.6 V versus Ag/AgCl with a wide linear range from 20 to 2500 µM (R(2) = 0.9978) and a low detection limit of 2.0 µM (signal/noise [S/N] = 3). This electrode can effectively analyze glucose concentration in human serum samples, avoiding interference, and is a promising non-enzymatic glucose sensor due to its low overpotential, high sensitivity, good selectivity and stability, fast response, and low cost.

Molecular and morphological characterization of the tapeworm Taenia hydatigena (Pallas, 1766) in sheep from Iran.

Although Taenia hydatigena is one of the most prevalent taeniid species of livestock, very little molecular genetic information exists for this parasite. Up to 100 sheep isolates of T. hydatigena were collected from 19 abattoirs located in the provinces of Tehran, Alborz and Kerman. A calibrated microscope was used to measure the larval rostellar hook lengths. Following DNA extraction, fragments of cytochrome c oxidase 1 (CO1) and 12S rRNA genes were amplified by the polymerase chain reaction method and the amplicons were subjected to sequencing. The mean total length of large and small hooks was 203.4 µm and 135.9 µm, respectively. Forty CO1 and 39 12S rRNA sequence haplotypes were obtained in the study. The levels of pairwise nucleotide variation between individual haplotypes of CO1 and 12S rRNA genes were determined to be between 0.3-3.4% and 0.2-2.1%, respectively. The overall nucleotide variation among all the CO1 haplotypes was 9.7%, and for all the 12S rRNA haplotypes it was 10.1%. A significant difference was observed between rostellar hook morphometry and both CO1 and 12S rRNA sequence variability. A significantly high level of genetic variation was observed in the present study. The results showed that the 12S rRNA gene is more variable than CO1.

Cryptosporidium spp. Infection in human and domestic animals.

BACKGROUND: Cryptosporidium spp. is a coccidian parasite infected humans and animals. Prevalence rate of Cryptosporidium spp. infection associated with is some parameters such as sampling, age, season, country and contact to domestic animals. This study aimed to determine Cryptosporidium spp. Infection in humans and some animals in rural areas of Shushtar district from Khuzestan Province, south- west of Iran. METHODS: In this study, Stool specimens were randomly collected from 45 cattle, 8 buffalos, 35 calves, 22 turkeys, 3 sheep, 2 geese as well as 62 humans in different seasons selected from rural areas of Shushtar district located in Khuzestan in the south- west of Iran from August 2009 to April 2011. The collected stool samples were examined by modified Ziehl-Neelsen staining method. RESULTS: Altogether, 68/115 (59.1%) domestic animals and 9/62 (14.5%) of humans were showed Cryptosporidium spp. infection in the study areas. CONCLUSION: In this study we found the high frequency of Cryptosporidium spp. infection in the studied areas.

Seroprevalence of human hydatidosis using ELISA method in qom province, central iran.

BACKGROUND: The objective of this study was to determine the prevalence of cystic echinococcosis (CE) in Qom Province, central Iran using ELISA test. METHODS: Overall, 1564 serum samples (800 males and 764 females) were collected from selected subjects by randomized cluster sampling in 2011-2012. Sera were analyzed by ELISA test using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. RESULTS: Seropositivity was 1.6% (25 cases). Males (2.2%) showed significantly more positivity than females (0.9%) (P= 0.03). There was no significant association between CE seropositivity and age group, occupation, and region. Age group of 30-60 years encompassed the highest rate of positivity. The seropositivity of CE was 2.1% and 1.2% for urban and rural cases respectively. Binary logistic regression showed that males were 2.5 times at higher risk for infection than females. CONCLUSION: Although seroprevalence of CE is relatively low in Qom Province, yet due to the importance of the disease, all preventive measures should be taken into consideration.

Fatal Case of Plasmodium vivax Malaria in a Splenectomized Patient.

Malaria is a major problem in tropical and sub-tropical countries, with high morbidity and mortality. Splenectomy makes patients more susceptible to serious bacterial and parasitic infections. We report for the first time in Iran a fatal case of Plasmodium vivax malaria, confirmed by microscopic and molecular (Semi-nested multiplex PCR) tests in a patient who had undergone splenectomy due to hemolytic anemia.

Enteric protozoan parasites in rural areas of bandar-abbas, southern iran: comparison of past and present situation.

BACKGROUND: The main goal was to address the prevalence of enteric protozoan parasites in rural areas of Bandar-Abbas, southern Iran and to compare the results with the only conducted study in 1978. METHODS: This descriptive study was performed from 2009 through 2010 on the 565 fecal samples. Formalin-ether concentration technique was performed and the analysis was carried out using Chi-square test in SPSS software version 13.5. Finally, the comparison of our results with the only previous study which was accomplished by Sheiban and Rezaeian in 1978 was done. RESULTS: The overall prevalence of the protozoan parasites was 48.8%. However, the prevalence of pathogen parasites was 23%. Previous research in 1978 showed 80.4% infectivity. The most protozoan parasites were Blastocystis hominis (25.53%), Giardia lamblia (17.2%) and Entamoeba coli (15.95%). Previous study in 1978 found Entamoeba coli as the most common protozoa. Our finding revealed that the rate of single infectivity was much higher compared to previous research. The most frequency of infection was in children. CONCLUSION: The remarkable decrease of protozoan parasites is mainly due to progress in health care in the villages; however more effort should be done with the goal of eradicating infectious agents.

Isolation of free-living amoebae from sarein hot springs in ardebil province, iran.

BACKGROUND: Free-living amoebae (FLA) are a group of ubiquitous protozoan, which are distributed in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran. METHODS: Seven recreational hot springs were selected in Sarein City and 28 water samples (four from each hot spring) were collected using 500 ml sterile plastic bottles during three month. Filtration of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also performed using sequencing based method. RESULTS: Overall, 12 out of 28 (42.9%) samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebae were found in one (3.6%) and 11 (39.3%) samples, respectively. Sequence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T(4) genotype corresponding to A. castellanii. CONCLUSION: Contamination of hot springs to FLA, such as Acanthamoeba T(4) genotype (A. castellanii) and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence.

A Sensitive and Specific PCR Based Method for Identification of Cryptosporidium Sp. Using New Primers from 18S Ribosomal RNA.

BACKGROUND: The main goal of the present study was to develop a new sensitive and specific PCR based method for Identification of Cryptosporidium sp. using novel primers from 18S ribosomal RNA. Cryptosporidiosis in high-risk host groups particularly in neonates and immuno-compromised individuals may result in death. To the best of our knowledge this is the first study regarding develop a new PCR based method to diagnose the cryptosporidiosis in Iran. METHODS: A total of 850 human fecal samples from patients clinically suspected to cryptosporidiosis and 100 healthy and diarrheic cattle stool specimens were collected. The simplified formol-ether concentration method was carried out for all samples. They were then examined microscopically by modified Ziehl-Neelsen staining method. Total DNA was extracted by QIA amp DNA stool mini kit. PCR and nested-PCR was carried out by using designed primers. RESULTS: Twenty nine cases of cryptosporidiosis infection in human and 30 samples from cattle microscopically were positive. The described primary and nested PCR method could detect all Cryptosporidium positive samples from human and cattle. Regards to suspected negative samples in primary PCR examination, the Nested PCR could approve two more positive results. Furthermore, Nested PCR analysis was able to detect one more case which was negative in both microscopically examination and primary PCR. Specificity of the test was 100%. Sensitivity of Nested PCR in comparison to our gold standard; microscopy after Ridley concentration modified ziehl-Neelsen, was 100%. CONCLUSION: Our developed PCR based method by using new primers devised from 18S ribosomal RNA revealed the ability for identification of the Cryptosporidium species such as C. parvum and C. huminis with high specificity and sensitivity.

Molecular Identification and Sequencing of Mannose Binding Protein (MBP) Gene of Acanthamoeba palestinensis.

BACKGROUND: Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. palestinensis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. Mannose Binding Protein (MBP) is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize MBP gene in pathogenic Acanthamoeba isolates such as A. palestinensis. METHODS: This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008. A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using phenol-chloroform method. PCR reaction and amplification were done using specific primer pairs of MBP. The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank. RESULTS: A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the obtained sequence showed 81% similarity with the available MBP gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895, CONCLUSION: MBP is known as the most important factor in Acanthamoeba pathogenesis cascade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people.

Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar.

BACKGROUND: We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. METHODS: We developed a single PCR method for this purpose. After investigation of GenBank, primer pairs were designed from highly conserved regions of cysteine proteinase (CP5) gene. The primers were utilized in PCR using isolated genomic DNA template of E. histolytica and the PCR products were then sequenced. The same primer and method for PCR was used for isolated genomic DNA template of E. dispar. RESULTS: A fragment of about 950 bp was isolated in PCR by using DNA from E. histolytica, however, no banding pattern was produced by using the same primers for E. dispar. We characterized CP5 gene at molecular level in E. histolytica isolates from 22 positive; including 20 non-dysentery samples isolated from both cities as well as two dysentery samples isolated only from Tabriz. Nucleotide sequence comparison in gene data banks (NCBI, NIH) revealed significant homology with CP5 gene in E. histolytica isolates CONCLUSION: We developed a PCR method, which could detect simply and rapidly E. histolytica by amplifying a specific PCR fragment.

Intestinal protozoa in wild boars (Sus scrofa) in western Iran.

A total of 12 gastrointestinal tracts of wild boars (Sus scrofa) from western Iran (Luristan) were examined for protozoan infection between September 2000 and November 2001. Of 12 boars examined, 67% harbored one or more species of the following protozoa: Balantidium coli (25%), Tritrichomonas suis (25%), Blastocystis sp. (25%), Entamoeba polecki (17%), Entamoeba suis (8%), Iodamoeba butschlii (17%), and Chilomastix mesnili (8%). Four of these protozoan species also are reported in humans, and persons living in rural areas where wild boars are abundant should take precaution to avoid infection.