Generation of Zebrafish Models of Human Retinitis Pigmentosa Diseases Using CRISPR/Cas9-Mediated Gene Editing System.

Generating animal models can explore the role of new candidate genes in causing diseases and the pathogenicity of a specific mutation in the underlying genes. These animals can be used to identify new pharmaceutical or genetic therapeutic methods. In the present experiment, we developed a rpe65a knock out (KO) zebrafish as a retinitis pigmentosa (RP) disease model. Using the CRISPR/Cas9 system, the rpe65a gene was KO in zebrafish. Two specific single-guide RNAs (sgRNAs) were designed for the zebrafish rpe65a gene. SgRNAs were cloned into the DR274 plasmid and synthesized using in vitro transcription method. The efficiency of Ribonucleoprotein (synthesized sgRNA and recombinant Cas9) was evaluated by in vitro digestion experiment. Ribonucleoprotein complexes were microinjected into one to four-celled eggs of the TU zebrafish strain. The effectiveness of sgRNAs in KO the target gene was determined using the Heteroduplex mobility assay (HMA) and Sanger sequencing. Online software was used to determine the percent of mosaicism in the sequenced samples. By examining the sequences of the larvae that showed a mobility shift in the HMA method, the presence of indels in the binding region of sgRNAs was confirmed, so the zebrafish model for RP disease established. Zebrafish is an ideal animal model for the functional study of various diseases involving different genes and mutations and used for evaluating different therapeutic approaches in human diseases. This study presents the production of rpe65a gene KO zebrafish models using CRISPR/Cas9 technology. This model can be used in RP pathophysiology studies and preclinical gene therapy experiments.

Green synthesis of nano-liposomes containing Bunium persicum and Trachyspermum ammi essential oils against Trichomonasvaginalis.

BACKGROUND: Trichomonas vaginalis, a parasitic flagellated protozoan, is one of the main non-viral sexually transmitted diseases worldwide. Treatment options for trichomoniasis are limited to nitroimidazole compounds. However, resistance to these drugs has been reported, which requires the development of new anti-Trichomonas agents that confer suitable efficacy and less toxicity. METHODS: In the present work, we assessed the effectiveness of the liposomal system containing essential oils of Bunium persicum and Trachyspermum ammi against T. vaginalis in vitro. The chemical composition of B. persicum and T. ammi were analyzed using gas chromatography-mass spectrometry (GC-MS). Liposomal vesicles were prepared with phosphatidylcholine) 70%) and cholesterol)30%) using the thin-film method. The essential oils of B. persicum and T. ammi were loaded into the liposomes using the inactive loading method. Liposomal vesicles were made for two plants separately. Their physicochemical features were tested using Zeta-Sizer, AFM and SEM. The anti-Trichomonas activity was determined after 12 and 24 h of parasite cultures in TYI-S-33 medium. RESULTS: After 12 and 24 h of administration, the IC50 of the B. persicum essential oil nano-liposomes induced 14.41 µg/mL and 45.19 µg/mL, respectively. The IC50 of T. ammi essential oil nano-liposomes induced 8.08 µg/mL and 25.81 µg/mL, respectively. CONCLUSIONS: These data suggested that nano-liposomes of the essential oils of B. persicum and T. ammi may be a promising alternative to current treatments for Trichomonas infection.

Cytogenetic Studies of 608 Couples with Recurrent Spontaneous Abortions in Northeastern Iran.

BACKGROUND & OBJECTIVE: One of the major genetic causes of recurrent spontaneous abortions is parental chromosomal abnormalities. The objectives of the study were to determine, compare and analyze the incidence and distribution of chromosomal abnormalities in couples with recurrent miscarriages from Northeastern Iran. METHODS: This study was conducted at Ghaem Hospital, Mashhad, Iran. We evaluated karyotype results of 608 couples with history of recurrent spontaneous abortion. The standard method was used for culturing peripheral venous blood lymphocytes. RESULTS: Chromosome aberrations were detected in 43 patients (3.54%), including 25 females and 18 males. Structural chromosomal abnormality was detected in 40 cases, including balanced translocations (25 cases), robertsonian translocations (4 cases), inversions (10 cases) and numerical chromosome aberrations (3 cases). Polymorphic variants were observed in 22 individuals. CONCLUSION: The frequency of chromosomal abnormalities in couples with Recurrent Spontaneous Abortion (RSA) in our study is 3.54%. Reciprocal translocation, pericentric inversions, robertsonian translocations, and numerical abnormality observed among couples who had experienced recurrent spontaneous abortions and that these couples might benefit from cytogenetic analysis.

Seroprevalence of toxoplasmosis in Kermanshah City, west of Iran.

It is estimated that one-third of the world's population is infected with Toxoplasma gondii. The purpose of this study was to evaluate the latest status of toxoplasmosis seroprevalence in the general population and pregnant women in the west of Iran. This retrospective cross-sectional study was conducted in 2018. Accordingly, data associated with serodiagnosis of toxoplasmosis, age, sex, anti-toxoplasmosis IgG and IgM, and pregnancy status in women were collected from 6 health centers of Kermanshah City, the west of Iran, during 2016-2017. In total, 1228 people referred to the health centers in Kermanshah City. Of 1228 people, 359 (29.23%) individuals were seropositive for toxoplasmosis, of them 294 (81.89%) individuals were seropositive only for IgG, and 65 (18.11%) individuals were both IgG and IgM seropositive. The seropositivity in men was 29.3% (n = 63), in women was 29.2% (n = 296), and in pregnant women was 25.9% (n = 44). All individuals were examined using ELISA kit. This study showed that the prevalence of this disease in the west of Iran has been decreased in comparison with the previous studies. Therefore, regular epidemiological studies of in different regions seem to be necessary in order to conclude on the decrease or increase trend of this disease in an area.

Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes.

Peganum harmala, a well-known medicinal plant, has been used for several therapeutic purposes as it contains numerous pharmacological active compounds. Our study reported an anti-parasitic activity of P. harmala seed extract against Acanthamoeba triangularis. The stress induced by the extract on the surviving trophozoites for Acanthamoeba encystation and vacuolization was examined by microscopy, and transcriptional expression of Acanthamoeba autophagy-related genes was investigated by quantitative PCR. Our results showed that the surviving trophozoites were not transformed into cysts, and the number of trophozoites with enlarged vacuoles were not significantly different from that of untreated control. Molecular analysis data demonstrated that the mRNA expression of tested AcATG genes, i.e., ATG3, ATG8b, and ATG16, was at a basal level along the treatment. However, upregulation of AcATG16 at 24 h post treatment was observed, which may indicate an autophagic activity of this protein in response to the stress. Altogether, these data revealed the anti-Acanthamoeba activity of P. harmala extract and indicated the association of autophagy mRNA expression and cyst formation under the extract stress, representing a promising plant for future drug development. However, further identification of an active compound and a study of autophagy at the protein level are needed.

A novel delivery of curcumin by the efficient nanoliposomal approach against Leishmania major.

Several side effects and drug resistance accompany the current therapies for Leishmaniasis. Nanoliposomal curcumin is applied as a new therapy approach instead of current therapy. In this study, nanoliposomal curcumin was prepared using thin-film hydration method and characterized based on encapsulation efficiency, size, and zeta potential. Curcumin was successfully loaded into nanoliposomes with an encapsulation efficiency of 92%. The surface charge of the nanoparticle was neutral, and the size of nanoparticle was 176.5 nm. Nanoliposomal curcumin is in spherical shape without any agglomeration. Cell viability assay was performed on HFF cell line to show biocompatibility of liposome nanoparticles. Anti-Leishmanial effect of different concentrations of liposomal curcumin (0.05-30 µg mL-1) and amphotericin B (25 µg mL-1) were studied on Leishmania major [MRHO/IR/75/ER] at various hours (24, 48, and 72) using hemocytometer technique. Nanoliposomal curcumin inhibitory concentration (IC50) at hours 24, 48, and 72 were 6.41, 3.8, and 2.33 µg mL-1, respectively. As prepared nanoliposomal curcumin showed a significant antileishmanial effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigations.

Amoebicidal activity of Cassia angustifolia extract and its effect on Acanthamoeba triangularis autophagy-related gene expression at the transcriptional level.

Cassia angustifolia Vahl. plant is used for many therapeutic purposes, for example, in people with constipation, skin diseases, including helminthic and parasitic infections. In our study, we demonstrated an amoebicidal activity of C. angustifolia extract against Acanthamoeba triangularis trophozoite at a micromolar level. Scanning electron microscopy (SEM) images displayed morphological changes in the Acanthamoeba trophozoite, which included the formation of pores in cell membrane and the membrane rupture. In addition to the amoebicidal activity, effects of the extract on surviving trophozoites were observed, which included cyst formation and vacuolization by a microscope and transcriptional expression of Acanthamoeba autophagy in response to the stress by quantitative polymerase chain reaction. Our data showed that the surviving trophozoites were not transformed into cysts and the trophozoite number with enlarged vacuole was not significantly different from that of untreated control. Molecular analysis data demonstrated that the mRNA expression of AcATG genes was slightly changed. Interestingly, AcATG16 decreased significantly at 12 h post treatment, which may indicate a transcriptional regulation by the extract or a balance of intracellular signalling pathways in response to the stress, whereas AcATG3 and AcATG8b remained unchanged. Altogether, these data reveal the anti-Acanthamoeba activity of C. angustifolia extract and the autophagic response in the surviving trophozoites under the plant extract pressure, along with data on the formation of cysts. These represent a promising plant for future drug development. However, further isolation and purification of an active compound and cytotoxicity against human cells are needed, including a study on the autophagic response at the protein level.

Cytogenetic Study of Patients with Primary Amenorrhea in the Northeast of Iran.

BACKGROUND & OBJECTIVE: Primary amenorrhea refers to the absence of menstruation in females of reproductive by age 16 when the development of secondary sexual characteristics is evident (breast development, pubic hair) or by age 14 when there are no secondary sexual characteristics are present. Primary amenorrhea can occur in several quite different reasons. Common hormonal causes of primary amenorrhea include constitutional delay, hypothalamic or pituitary disorders, chronic systemic disease, and primary ovarian insufficiency, some endocrine gland disorders, and other causes. Previous studies suggested that chromosomal abnormality is the second most common cause of amenorrhea. This report aims to measure the prevalence of the chromosomal abnormality in primary amenorrhea (PA) patients in the northeast of Iran. METHODS: Chromosomal study was carried out on 200 patients with clinical features. The standard method for culturing peripheral venous blood lymphocyte was to prepare metaphase chromosomes and perform routine GTG band analysis. RESULTS: The results revealed that 71% of PA had normal female karyotype (46,XX) and 29% showed different chromosomal abnormalities. The chromosomal abnormalities can be categorized into seven primary groups with or without mosaicism. 1- The most common karyotype was X chromosome aneuploidy (10.5%, n=21), 2- Male karyotype with or without structural abnormality of Y chromosome (5.5 %, n=11), 3- Mosaicism of turner karyotype and structural anomalies of X chromosome (4%, n=8), 4- Structural anomalies of the X chromosome (3.5%, n=7), 5- Mosaicism of turner karyotype and normal karyotype (3%, n=6), 6- Mosaicism of turner karyotype and male karyotype (1.5%, n=3) and 7- Super female karyotype (1%, n=2). CONCLUSION: The present study has emphasized that early cytogenetic and timely investigation can be necessary for the evaluation of primary amenorrhea.

In vitro evaluation of nanoliposomes berberine chloride against Leishmania major promastigotes.

Leishmaniosis caused by Leishmania major is one of the main infectious diseases that infected populations in developing countries around the world. We assessed the effectiveness of berberine chloride nanoliposomes (BcNLs) against L. major promastigotes in vitro. Nanoliposomal berberine chloride was prepared using thin film hydration method and characterized based on encapsulation efficiency, size and zeta potential. Anti-Leishmania effect of different concentrations (0.05-60 µg/ml) of BcNLs as studied in L. major (MRHO/IR/75/ER) at 24, 48 and 72 h using the hemocytometer technique. Berberine chloride was successfully loaded into nanoliposomes with encapsulation efficiency of 85.54%. The surface charge of nanoparticle is neutral and the morphology of nanoliposomal berbrine chloride is spherical without any agglomeration. Cell viability assay was performed on HFF cell line to show biocompatibility of liposome nanoparticles. IC50 of BcNPs at 24, 48 and 72 h against L. major were found to be 7.6, 5.96 and 3.19 µg/ml, respectively. BcNLs showed a significant anti-Leishmania effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigation. The results showed that the BcNLs agent is effective against L. major promastigotes and may be a promising alternative to current treatments.

Blood Parasites in Domestic Birds in Central Iran.

Parasites may affect the dynamics of bird populations. Plasmodium, Leucocytozoon and Haemoproteus are well-known avian haematozoa that can trigger decreased productivity and high mortality in domesticated birds. In this study, we evaluated the prevalence of avian blood parasites (Plasmodium, Leucocytozoon and Haemoproteus) against 335 birds of 8 species in the Yazd province in central Iran. To detect blood parasites, Giemsa-stained blood smears were prepared. Of the birds, 11.64% (39/335) were infected with at least one parasite genus, particularly Haemoproteus (32.6%; 23/335). The total prevalence values for Plasmodium, Haemoproteus and Leucocytozoon were 1.7, 6.8 and 2.9%, respectively. Plasmodium had lower prevalence rates of 1.7% (6/335). Among birds, pigeons, hens and ducks have the highest prevalence of Haemoproteus, Leucocytozoon and Plasmodium parasites at 1.7%, 6.8% and 2.9%, respectively. Results from this research extend our knowledge on the incidence of avian blood parasites in domesticated birds living in central Iran. The overall low incidence of avian blood parasites in birds was found in the Yazd province, Iran.

Isolation and Molecular Identification of Leishmania spp. Agents in Patients with Cutaneous Leishmaniasis in Yazd Province, Endemic Region of Central Iran.

BACKGROUND: Cutaneous Leishmaniasis (CL) is a major health problem in many parts of Iran. Many methods have been introduced for detection and identification of Cutaneous Leishmaniasis. The purpose of this study was isolation and molecular identification of Leishmania spp. agents in patients with CL from endemic region of central Iran. In this study, one of the main loci of central Iran named Yazd will be assessed CL identification using PCR-RFLP. METHODS: For this cross-sectional study, sampling was done from 372 suspicious patients with CL who referred to Health Centers of Yazd Province from 2016 to 2017. After collection samples of patients, DNA extraction was done from samples on slides. Genus detection was done using specific primers by PCR. RFLP analysis was done for species identification. RESULTS: Out of 372 samples, 159 samples were positive using PCR based method. Out of 159 samples, 87 (54.7%) L. major and 72 (45.3%) L. tropica were identified using RFLP analysis. The number of lesions in each patient was different but 119 (74.8%) patients showed the number of 1-3 lesions, and more lesions (more than 10 lesions) was showed in 4 (2.5%) person. CONCLUSION: The CL found in Yazd province resulted from L. major and L. tropica as the agents of rural and urban types, respectively. The prevalence of L. major and L. tropica was almost the same. This indicated that control programs could be designed for treatment and vector and reservoir control.

Evaluation of the Anti-Leishmanial Effect of Recombinant Clostridium α-Toxin.

BACKGROUND: Leishmaniasis is an infectious disease common in tropical and subtropical regions caused by the genus Leishmania, which is transmitted by the bite of female sandflies. In this study, we evaluate the anti-leishmanial effect of recombinant Clostridium α-toxin protein alone and the combination with glucantime through in vitro and in vivo. MATERIALS AND METHODS: Production, expression, and purification of recombinant α-toxin were evaluated by SDS-PAGE and Western blotting techniques. The antileishmanial activities of the purified α-toxin plus and without glucantime were examined in vitro and in vivo. RESULTS: The results indicated successful expression of α-toxin as a 48 kDa band on SDS-PAGE and Western blot methods. Also, evaluation of α-toxin IC50 showed the strong fatal effect of it, and glucantime on medium proliferated Leishmania promastigotes at lower concentrations compared with glucantime or α-toxin alone. Moreover, in vivo surveys showed that at the end of treatment courses, the mean of lesion size diminished in glucantime plus α-toxin treated mice versus negative control groups (p < 0.001). Also, there was a significant difference in the parasite burden of the spleen and liver of the control versus the test groups (p < 0.001). CONCLUSION: The results showed recombinant α-toxin has synergistic effects with glucantime in destroying Leishmania parasites.

Frequency and Molecular Identification of Cryptosporidium Species among Immunocompromised Patients Referred to Hospitals, Central Iran, 2015-16.

BACKGROUND: The purpose of this study was to determine the prevalence and genotype of Cryptosporidium spp. in different groups of immunocompromised patients admitted to the referral hospitals in center of Iran during 2015-2016. METHODS: This cross-sectional study was conducted on 346 immunocompromised patients (HIV+/AIDS, Lymphoma, Leukemia and organ transplants) in referred hospitals from central parts of Iran including Isfahan, Markazi, Yazd and Chaharmahale Bakhtiari provinces. Stool samples were analyzed for Cryptosporidium species, modified Ziehl-Neelsen staining techniques followed by the semi-nested PCR and DNA sequencing methods. RESULTS: The total rate of Cryptosporidium spp. was 3.46% (12/346) in the patients, however, the prevalence of the parasite, was 4.6% (4/87) in HIV+/AIDS patients, 3.6% (6/168) in patients with blood malignancy and 2.1% (2/91) in organ transplant recipients. The SSU rRNA gene of Cryptosporidium spp. in all microscopic-positive samples was effectively amplified by the semi-nested PCR and DNA sequences, exposed the existence of two Cryptosporidium species, including C. hominis 91.6% (11/12) and C. parvum 8.3% (1/12). CONCLUSION: The predominance of C. hominis in the present study may be certifies the importance of anthroponotic transmission of cryptosporidiosis in center of Iran.

Production and characterization of recombinant human leukemia inhibitory factor and evaluation of anti-fertility effects of rabbit anti-rhLIF in Balb/c mice.

Human leukemia inhibitory factor (hLIF) is a cytokine of interleukin-6 family. This study aimed to evaluate the recombinant production rate of active hLIF by different vector-host systems under various conditions. Moreover, a rabbit polyclonal antibody (pAb) against recombinant hLIF (rhLIF) was produced and its anti-fertility effects were explored in Balb/c mice. Four different constructs including pET22b/hLIF, pET28b/hLIF, pET32b/hLIF and pColdI/hLIF were designed and transformed into BL21-(DE3), Rosetta-(DE3), Origami-(DE3) and Shuffle T7-(DE3) host cells. The expression level and proliferative effect of rhLIF were measured by SDS-PAGE and MTT assays, respectively. Rabbit pAb to rhLIF was produced and characterized using enzyme-linked immunosorbent assay and western blot techniques. The Balb/c mice were divided into two intervention and control groups. Then, they were intraperitoneally injected by purified rabbit anti-rhLIF and non-immunized rabbit pAb, respectively. After sacrifice on day 7, the number of implantation sites was counted. The rhLIF was successfully expressed by pET32b/hLIF and pColdI/hLIF vectors in all hosts with no significant difference in the rate of their expression. The rhLIF was purified and checked for activity. The results showed that it is functionally active and the produced anti-rhLIF pAb could specifically bind to commercial rhLIF. Passive immunization results showed that anti-rhLIF antibody completely inhibited fertility in all injected Balb/c mice compared to controls. Although previous studies showed expression of rhLIF using various methods, using different vector-host systems ensures us of successful biological active expression of it. The pAb against rhLIF could be a powerful tool for inducing in vivo infertility.

A novel mutation in USF1 gene is associated with familial combined hyperlipidemia.

BACKGROUND: Familial combined hyperlipidemia or FCHL is one of the most common genetic causes of hyperlipidemia and is associated with elevation of cholesterol, triglycerides or both, and increased serum apolipoprotein B (apoB). Linkage analysis and next generation sequencing have been successfully used for identifying rare genetic variants that have moderate-to-large effects. METHODS: We characterized a large pedigree from a proband identified following recruitment into the MASHAD study, in northeast Iran, with FCHL accompanied by early-onset coronary artery disease. We used linkage analysis for several candidate regions in previous studies such as 1q21-23, 11q23, and 8p, and then whole-exome sequencing to identify the disease-associated gene in this family. RESULTS: We identified a novel variant in the USF1 gene, leading to a substitution of a tryptophan for arginine at position 196. Arg196Trp co-segregated in all the affected family members in this pedigree with clinical syndrome and was not found in any unaffected family members of this pedigree, or in unrelated controls. CONCLUSIONS: We speculate that this mutation [Arg196Trp] in the USF1 gene might be associated with FCHL and early-onset coronary heart disease in this family. However, the substantial mechanism requires further investigation. These findings indicate that USF1 plays an important role in the biological pathways associated with lipid metabolism.

A Novel Strategy for Enhance Potentiation of Meglumine antimoniate against Leishmania major In Vitro.

BACKGROUND: We aimed to design a different method of drug delivery for increased transfer of the choice drug (meglumine antimoniate) within the host cells. Therefore, listeriolysin O (LLO), a bacterial product which is a member of pore-forming peptides was used as an enhancer factor with meglumine antimoniate in order to facilitate the transition of the drug across macrophage membrane. METHODS: LLO was produced in Isfahan University of Medical Sciences in 2016, by expressing the hlyA gene in Escherichia coli and purified using affinity chromatography. Cytotoxicity of the purified protein was investigated in an in vitro model of macrophage Leishmania infection. RESULTS: LLO was cytotoxic against murine macrophage cells (J774-A1) and amastigote forms of L. major (MRHO/IR/75/ER). It was less toxic to macrophages (CC50=2.56 µg ml-1 ±0.09) than to the parasites (IC50=1.72 µg ml-1 ±0.07). Moreover, noncytotoxic concentration of LLO (0.006 ug ml-1) potentiated the cytotoxicity induced by low dose concentration of meglumine antimoniate. Very little dose of meglumine antimoniate was needed when combined with the LLO (IC50=12.63 µg ml-1 ±0.13) in comparison with the cytotoxicity induced when the drug is used alone (IC50=46.17 µg ml-1 ±0.28). CONCLUSION: The combination of pore-forming proteins with anti-leishmanial agents could increase the advantage of anti-leishmanial drugs. Since lower concentrations of anti-leishmanial drugs can reduce undesirable side effects of chemotherapy trials carried out in animal models and then in humans with this system.

Prevalence and clinical features of Cryptosporidium infection in hemodialysis patients.

AIM: This cross-sectional study aims to assess the prevalence of Cryptosporidium in hemodialysis patients compared with healthy individuals in central Iran from August 2014 to January 2015. BACKGROUND: Cryptosporidiosis is a major cause of acute and persistent diarrhea with significant morbidity and mortality in immunocompromised patients such as those undergoing renal dialysis. METHODS: Three stool samples were collected from 330 hemodialysis patients and 150 healthy individuals on 3 consecutive days. The samples were screened for Cryptosporidium infection using formalin-ether sedimentation and modified Ziehl-Neelsen staining. Demographic variables as well as risk factors were recorded. RESULTS: Out of 330 dialysis patients and 150 healthy individuals, 10 (3%) and 1 (0.7%) were infected with Cryptosporidium, respectively. We found statistically significant differences between infection and place of residency, hygiene status, education level, diarrhea, and abdominal pain in the two groups (p<0.05). On the other hand, there was no relationship between infection and sex, contact with domestic animals, fever, vomiting, nausea, flatulence, anorexia, duration of dialysis and underlying disorders in the two groups. Also, there was a statistically significant difference between age and infection in hemodialysis patients (p=0.003). A higher infection rate was observed in patients under 20 years of age. CONCLUSION: Risk factors for Cryptosporidium infection must be controlled. We strongly recommended that stool samples from such patients, especially those with severe or prolonged diarrhea, should be examined with modified Ziehl-Neelsen staining for appropriate and timely treatment.

In vitro anti-leishmanial activity of Satureja hortensis and Artemisia dracunculus extracts on Leishmania major promastigotes.

Cutaneous leishmaniasis is one of the important skin diseases with diverse clinical manifestations. With an incidence of 0.7-1.3 million cases annually, this disease is often reported from six countries, including Iran. Accordingly, the purpose of this study was to evaluate the anti-leishmanial effect of the three plant hydroalcoholic extracts including fleawort (Plantago psyllium L.), savory (Satureja hortensis L.) and tarragon (Artemisia dracunculus L.) on Leishmania major promastigotes. The hydroalcoholic extract from each plant was extracted and its anti-leishmanial effect was evaluated in different concentrations (100-1000 µg/ml) and at various hours (24, 48 and 72 h). Savory herb inhibitory concentration 50 % (IC50) at 24, 48 and 72 h was 790.81, 398.11 and 298.42 µg/ml, respectively. In addition, tarragon herb IC50 at 24, 48 and 72 h was 962.03, 688.36 and 585.51 µg/ml, respectively. Moreover, the fleawort extract was showed the lowest effect, considering that its effect at the concentration of 1000 µg/ml was 48 % after 72 h (P > 0.05). Furthermore, the statistical analysis showed a significant difference for interaction between concentration and time regarding the tarragon and savory extracts with a P value of lower than 0.05. According to the results, the anti-leishmanial effect of the tarragon and savory extracts may make it possible to use them in the treatment of cutaneous leishmaniasis as a complementary or alternative therapy; however, further studies are necessary and should be evaluated in cell culture and in vivo conditions to confirm it.

Chromosomal analysis of couples with repeated spontaneous abortions in northeastern iran.

BACKGROUND: Cytogenetic study of reproductive wastage is an important aspect in determining the genetic background of early embryogenesis. Approximately 15 to 20% of all pregnancies in humans are terminated as recurrent spontaneous abortions (RSAs). The aim of this study was to detect chromosome abnormalities in couples with RSAs and to compare our results with those reported previously. MATERIALS AND METHODS: In this retrospective study, the pattern of chromosomal aberrations was evaluated during a six-year period from 2005 to 2011. The population under study was 728 couples who attended genetic counseling services for their RSAs at Pardis Clinical and Genetics Laboratory, Mashhad, Iran. RESULTS: In this study, about 11.7% of couples were carriers of chromosomal aberrations. The majority of abnormalities were found in couples with history of abortion, without stillbirth or livebirth. Balanced reciprocal translocations, Robertsonian translocations, inversions and sex chromosome aneuploidy were seen in these cases. Balanced reciprocal translocations were the most frequent chromosomal anomalies (62.7%) detected in current study. CONCLUSION: These findings suggest that chromosomal abnormalities can be one of the important causes of RSAs. In addition, cytogenetic study of families who experienced RSAs may prevent unnecessary treatment if RSA are caused by chromosomal abnormalities. The results of cytogenetic studies of RSA cases will provide a standard protocol for the genetic counselors in order to follow up and to help these families.

SEROPREVALENCE OF TOXOPLASMA GONDII INFECTION AMONG PATIENTS ADMITTED TO AL-ZAHRA HOSPITAL, ISFAHAN, IRAN.

BACKGROUND: Toxoplasma gondii (T. gondii) infection is one of the most common parasitic infections among humans and other warm-blooded animals worldwide. The aim of this study was to evaluate toxoplasmosis status in patients admitted to Al-Zahra hospital, Isfahan, Iran. METHODS: This cross-sectional study was conducted from October 2012 to January 2015. During this period, 716 patients referred to AI-Zahra hospital in Isfahan city, Iran, were studied to investigate the IgG and IgM antibodies against T. gondii using ELISA kit. The data were analysed by Chi-square and Fisher's exact tests. In addition, the relation of data with age and sex were also examined. RESULTS: Among 716 patients, 21 patients (2.9%) had positive IgM and 288 patients (40.2%) had positive IgG titer against T. gondii. Data analysis by Chi-square and Fisher's exact tests revealed that there was no significant relationship between IgG titer and age (p > 0.05). Additionally, there was no relationship between IgM titer and age (p > 0.05). The data showed that there was no relationship between IgG and IgM antibody titer and sex (p > 0.05). CONCLUSION: The prevalence of toxoplasmosis in Isfahan inhabitants seems fairly high but it can be concluded that the rate of seropositive patient is moderate comparing to other regions of country. Accordingly, the authors propose that all sensitive patients have to be tested for T. gondii antibody in order to prevent the consequences of disease.