Study of toxoplasmosis and toxocariasis in patients suffering from ophthalmic disorders using serological and molecular methods.

INTRODUCTION: Toxoplasma gondii is an intracellular protozoan parasite that can cause ocular toxoplasmosis with most complications such as retinal detachment. Toxocara parasite, round worm, found in dogs and cats appears as larva migrans in humans can cause serious ocular complications such as debilitating vision loss.In Khuzestan province, southwest of Iran, T. gondii infection has been reported to be significant but toxocariasis was rare. However, the frequency of ocular toxoplasmosis and toxocariasis has not been studied in this area. The aim of this study was to evaluate the ocular toxoplasmosis and ocular toxocariasis using serological and molecular methods. METHOD: In this case control study, 310 patients were identified by ophthalmologist as ocular toxoplasmosis and then 5 cc of venous blood samples were taken from each of them. Serum samples and buffy coat were prepared and ELISA was used to detect IgG and IgM anti-Toxoplasma antibodies and the molecular PCR was used to detect Toxoplasma DNA parasite in buffy coats. ELISA test was used to detect of IgG anti-Toxocara antibodies. RESULTS: Totally, for ocular toxoplasmosis, 130 (41.93%) of 310 patients were positive by ELISA, of them 121 (39%) IgG positive and nine (2.9%) IgM positive were diagnosed. Of 121 cases with IgG+, 119 (98.35%) were diagnosed with high IgG avidity indicating chronic phase of the infection. For ocular toxocariasis evaluation, antibodies against Toxocara were not detected in any of the samples. By PCR molecular method, 11 out of 310 patients (3.54%) had T. gondii DNA in the blood. In control, in total, 21 cases were detected positive by serology method, which showed a significant difference with the results of the case group(P < 0.05).By PCR method, only three cases showed positive which also indicated significant difference with result of case group (3 vs 9) (P < 0.05). In the control group, also no anti-toxocara antibodies were found. CONCLUSION: It can be concluded that T. gondii in Khuzestan province as the etiologic agent of ocular toxoplasmosis and physicians should consider diagnostic methods for identifying the infection when they visit the patients.

Toxoplasma gondii and Toxocara spp. contamination in university area.

Toxoplasma gondii and Toxocara spp. zoonotic infections may cause severe systemic and ocular illness in infected individuals. Cats play a significant role in environmental contamination and the transmission of parasites. The goal of the present study was to investigate the prevalence of Toxoplasma gondii (T. gondii) and Toxocara spp. infection among stray cats at Ahvaz Jundishapur University of Medical Sciences campus. The current descriptive study began with the collection of 170 fresh cat faecal samples from various sites in the Ahvaz Jundishapur University of Medical Sciences area. Sheather's sugar flotation method was applied to all specimens, and parasites were identified and examined microscopically. Next, a nested-PCR assay, sequencing, and real-time PCR with high-resolution melting curve (HRM) analysis were performed. In this study, out of 170 cat faecal samples microscopically evaluated, 8 (4.70%) and 37 (21.76%) were infected with T. gondii oocysts and Toxocara eggs, respectively. Using nested PCR, 8 out of 170 samples (4.70%) were found to be infected with T. gondii. HRM analysis showed that all isolates could be classified into three genetic lineages. Considerable prevalence, exceeding 50% for Toxocara and surpassing 25% for Toxoplasma in certain instances, along with genetic diversity, was observed in the present study. Hence, it is suggested that all individuals, including kindergarten children, students, employees, workers, and pregnant women who are in contact with their surroundings, take the necessary precautions.

Toxoplasma gondii molecular detection and phylogenetic analysis in hemodialysis patients from Khuzestan, Southwest Iran.

BACKGROUND: The diagnosis and genetic characterization of Toxoplasma gondii (T. gondii) infection can make a significant influence to the prevention of the dangerous consequences of toxoplasmosis, particularly in immunocompromised people. OBJECTIVE: The aim of this investigation was to assess the frequency and genotyping of T. gondii in blood samples of patients with hemodialysis. MATERIALS AND METHODS: In the current investigation, a total of 379 blood samples were taken from subjects with hemodialysis who were referred to teaching hospital of Ahvaz in the southwest of Iran. The samples were evaluated using the Nested PCR by targeting the B1 gene, and then, sequencing and phylogenetic tree were constructed. RESULTS: T. gondii DNA was found in 112 (29.55%) of the blood samples by Nested PCR. Amplicons from T. gondii revealed high identity with GenBank sequences. The phylogenetic analysis revealed that all sequences were closely related to Type I of T. gondii. CONCLUSION: Because of the high incidence of toxoplasmosis with type I prevalent in hemodialysis patients, we recommend a systematic screening for toxoplasmosis to carry out for monitoring the possible dissemination of toxoplasmosis during hemodialysis.

Adipose-Derived Stem Cells' Secretome Attenuates Lesion Size and Parasite Loading in Leishmaniasis Caused by Leishmania Major in Mice.

Background: Stem cell-derived secretome (SE) released into the extracellular space contributes to tissue repair. The present study aimed to investigate the impact of isolated secretome (SE) from adipose-derived mesenchymal stem cells (ASCs) on Leishmania major (L. major) lesions in BALB/c mice. Methods: This experimental study was conducted at Ahvaz University of Medical Sciences (Ahvaz, Iran) in 2021. Forty female BALB/c mice were infected with stationary phase promastigotes through intradermal injection in the bottom of their tail and randomly divided into four groups (n=10 per group). The mice were given SE (20 mg/mL), either alone or in combination with Glucantime (GC, 20 mg/mL/Kg), meglumine antimoniate (20 mg/mL/Kg) for the GC group, and phosphate-buffered saline (PBS) for the control group. After eight weeks, the lesion size, histopathology, the levels of Interleukin 10 (IL-10), and Interleukin 12 (IL-12) were assessed. For the comparison of values between groups, the parametric one-way ANOVA was used to assess statistical significance. Results: At the end of the experiment, the mice that received SE had smaller lesions (4.56±0.83 mm versus 3.62±0.59 mm, P=0.092), lower levels of IL-10 (66.5±9.7 pg/mL versus 285.4±25.2 pg/mL, P<0.001), and higher levels of IL-12 (152.2±14.2 pg/mL versus 24.2±4.4 pg/mL, P<0.001) than the control. Histopathology findings revealed that mice treated with SE had a lower parasite burden in lesions and spleen than the control group. Conclusion: The current study demonstrated that ADSC-derived SE could protect mice infected with L. major against leishmaniasis.

In vitro activity of green tea extract against Leishmania major promastigotes.

BACKGROUND: Systemic and topical treatment options against Leishmaniasis are limited to a few drugs with inconsistent efficacy and unacceptable side effects and none of them is suitable for all forms of the disease. OBJECTIVE: The aim of this study was to search the in vitro activity of green tea extract against L. major promastigotes and compare it with glucantime. METHODS: Extract was prepared by percolation method. The extract was dried and dissolved in DMSO 1% solvent. Leishmania major promastigotes treated with 6 concentrations (3, 6, 12, 24, 48, 96 mg/ml) of the extract. As control positive group glucantime 85 mg/ml and additional untreated control group were included in this study. All cultures were performed in triplicate. The promastigotes were also counted and their flagellate's motilities were assessed microscopically. RESULTS: Ethanolic extract of green tea showed significant leishmanicidal activity against L. major promastigotes in different concentrations. Notably there was a concordance in anti-leishmanial effect of the ethanol extract with the increasing of the dosage (3, 6, 12, 24, 48, 96 mg/ml). In comparison with glucantime the mean alive promastigotes in 12 mg/ml concentration of green tea was almost as same as 85 mg/ml glucantime and higher green tea extract concentrations were higher effective than glucantime. CONCLUSION: Our study revealed a novel pharmacological activity against promastigotes of L. major and suggests that green tea extract has the potential of being used in leishmaniasis but more studies are needed to find out its activity against amastigote and appropriate route of application.

The in vitro anti-Leishmania Effect of Zingiber officinale Extract on Promastigotes and Amastigotes of Leishmania major and Leishmania tropica

Objective: Recently, the use of pentavalent antimony compounds for Leishmaniasis treatment has been associated with disease recurrence, drug resistance, and severe side effects. Therefore, there is a need to develop alternative treatment strategies. This study investigates the in vitro effects of Zingiber officinale on promastigotes and amastigotes of Leishmania major and Leishmania tropica. Methods: Promastigotes and amastigotes of Leishmania major and Leishmania tropica were cultured and mass-produced in an RPMI1640 medium enriched with other necessary compounds. The MTT colorimetric method and calculating the IC50 value were used to evaluate the anti-leishmania activity of hydroalcoholic extract of Zingiber officinale. Results: The hydroalcoholic extract of Zingiber officinale inhibited the growth of Leishmania major and Leishmania tropica promastigotes in 24, 48, and 72 hours after in vitro incubation. The IC50 of hydroalcoholic extract of Zingiber officinale was 56 µg/mL for Leishmania major and 275 µg/mL for Leishmania tropica promastigotes after 72 hours. The IC50 of hydroalcoholic extract of Zingiber officinale was 75 µg/mL for Leishmania major and 325 µg/mL for Leishmania tropica amastigotes after 72 hours. Conclusion: The results showed that hydroalcoholic extract of Zingiber officinale has cytotoxicity properties, and Leishmania tropica has a higher resistance to hydroalcoholic extract of Zingiber officinale than Leishmania major. Further research is recommended.

Study of toxoplasmosis in type 2 diabetic patients using ELISA and B1 nested-PCR methods.

Toxoplasma gondii (T. gondii) is a ubiquitous, opportunistic organism, which actually infects all warmblooded animals. Diabetes is a silent, irritating metabolic disorder among which type 2 diabetes includes over 90% of cases globally. This case-control study was aimed to detect T. gondii infection in type 2 diabetic patients in Khuzestan province, southwest of Iran. Serological enzyme-linked immunosorbent assay (ELISA) and molecular polymerase chain reaction (PCR) method targeting B1 gene were employed to comparatively detect the parasitic infection among 377 diabetic patients and 200 non-diabetic subjects during 2016-2018. Considerably higher anti-T. gondii antibodies were determined in case group 44.29% (167/377), than in control group 19% (38/200) (P<0.05). Among diabetic patients, 153 (40.58%) were seropositive for IgG and 14 (3.71%) were seropositive for IgM, while 35 (17.5%) and 3 (1.5%) healthy people were seropositive regarding IgG and IgM, respectively (P<0.05). By nested PCR, B1 gene was identified in 36 out of 167 (21.55%) and 5 out of 38 (13.15%) of the seropositive samples of case and control groups, respectively. The prevalence of anti-T. gondii antibodies and DNA in diabetic patients was significantly higher than in non-diabetic patients. Thus, it could be recommended to routinely evaluate the chronicity of the infection in diabetic patients.

Miltefosine-induced apoptotic cell death on Leishmania major and L. tropica strains.

The aim of this study was to assess the cytotoxic effects of various concentrations of miltefosine on Leishmania major (MRHO/IR/75/ER) and L. tropica (MHOM/IR/02/Mash10) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Also, 50% effective doses (ED50) for L. major and L. tropica amastigotes were also determined. Annexin-V FLUOS staining was performed to study the cell death properties of miltefosine using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. In both strains of L. major and L. tropica, miltefosine induced dose-dependent death with features of apoptosis, including cell shrinkage, DNA laddering, and externalization of phosphatidylserine. The IC50 was achieved at 22 µM and 11 µM for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 µM and 4.2 µM, respectively. Our results indicate that miltefosine induces apoptosis of the causative agent of cutaneous leishmaniasis in a dose-dependent manner. Interestingly, L. major did not display any apoptotic changes when it was exposed to miltefosine in concentrations sufficient to kill L. tropica.

Detection of Acute and Chronic Toxoplasma gondii Infection among Women with History of Abortion in the Southwest of Iran.

BACKGROUND: Toxoplasma gondii (T. gondii) is one of the most common intracellular protozoan parasites, which can infect humans and a wide range of mammals and birds. The current study is aimed at investigating the occurrence of T. gondii infection in women with a history of abortion in Khuzestan, Iran. MATERIALS AND METHODS: A total of 480 women with an abortion history, as well as 200 pregnant women with a normal delivery, were examined in this study. The blood, placenta, and umbilical cord blood samples were assessed by the enzyme-linked immunosorbent assay (ELISA) and nested-polymerase chain reaction (PCR) assay. RESULTS: Based on the results of ELISA assay, the prevalence of toxoplasmosis was 30.83% in women with a history of abortion (25.62% with T. gondii IgG and 5.20% with T. gondii IgM). According to the IgG avidity test, 60.16% of IgG-positive samples showed high avidity, while 27.64% showed low avidity. On the other hand, the prevalence of toxoplasmosis in women with a normal delivery was 23% (21.5% with T. gondii IgG and 1.5% with T. gondii IgM). According to the IgG avidity test, 81.39% of these women showed high avidity, while only 4.65% showed low avidity. Based on the nested-PCR method, T. gondii DNA was detected in 14.18% of blood samples, 4.69% of placental samples, and 1.34% of umbilical cord samples, collected from 148 seropositive women with a history of abortion. Besides, using this method, the parasite DNA was identified in 4.34% of blood samples, collected from 46 seropositive women with a normal delivery, but not in any of the umbilical cord or placenta samples. CONCLUSION: The present results showed that T. gondii infection contributes to abortion in Khuzestan Province, Iran. Therefore, it is essential to investigate toxoplasmosis in pregnant women, especially in those who are seronegative, using molecular and serological methods and inform them about their disease and the associated risks.

Prevalence of Toxoplasma gondii and Toxocara canis Among Myositis Patients in the Southwest of Iran.

INTRODUCTION: Parasitic myositis is caused by some parasites, such as Toxoplasma gondii) T. gondii (and Toxocara canis (T. canis). So, the aim of the study was to evaluate the prevalence T. gondii and T. canis in patients with myositis and healthy individuals. METHODS: A total of 108 samples were randomly selected as the control (54 healthy individuals) and test (54 myositis patients) groups. IgG and IgM antibodies (Ab) against T. gondii and IgG Ab against T. canis were measured by the enzyme-linked immunosorbent assay (ELISA). The detection of chronic and acute toxoplasmosis was performed by the ELISA IgG avidity. The presence of T. gondii in the blood was evaluated using the nested polymerase chain reaction (nested-PCR). RESULTS: Of 108, 33 (30.6%) cases were positive for IgG against T. gondii that 19 (35.2%) and 14 (25.9%) were observed in myositis patients and healthy individuals, respectively (P=0.296). Of 19 positive cases, 12 (63.2%) and 7 (36.8%) cases were detected as chronic and acute toxoplasmosis, respectively, while, all positive cases in the control group had chronic toxoplasmosis (P=0.013). One (1.9%) sample was positive for anti- T. gondii IgM and two (3.7%) samples were positive for IgG against T. canis by the ELISA that these positive cases were observed only in myositis patients (P=1.000 P=0.495, respectively). B1 T. gondii gene was amplified in 12 (63.2%) and 1 (7.1%) in myositis patients and healthy subjects (P=0.001). CONCLUSION: Our findings showed that there was a relatively high prevalence of acute toxoplasmosis in myositis patients in comparison with the control subjects in the Southwest of Iran.

Spermatogonia apoptosis induction as a possible mechanism of Toxoplasma gondii-induced male infertility.

OBJECTIVES: The protozoan Toxoplasma gondii as an intracellular protozoan is widely prevalent in humans and animals. Infection generally occurs through consuming food contaminated with oocysts and tissue cysts from undercooked meat. The parasite is carried in sexual fluids like semen but there is little information about the effect of T. gondii on the male reproductive system. In this study, we examined the effect of T. gondii tachyzoites on apoptosis induction in type B spermatogonia (GC-1) cells. MATERIALS AND METHODS: Fresh tachyzoites taken of infected BALB/c mice, GC-1 spg cells were infected with increasing concentrations of tachyzoites of T. gondii, then apoptotic cells were identified and quantified by flow cytometry. The genes associated with apoptosis were evaluated by RT2 Profiler PCR Array. RESULTS: PCR array analysis of 84 apoptosis-related genes demonstrated that 12 genes were up-regulated at least 4-fold and that one gene was down-regulated at least 2-fold in the T. gondii infection group compared with levels in the control group. The number of genes whose expression had increased during the period of infection with T. gondii was significantly higher than those whose expressions had decreased (18 versus 1) and Tnfrsf11b had the highest rate of gene expression. CONCLUSION: T. gondii induce in vitro apoptosis of GC-1 spg cells. This effect shows a trend of concentration-dependent increase so that with an increase in the ratio of parasite burden to spermatogonial cells, in addition to an increase in the number of genes whose expression has changed, the fold of these changes has increased as well.

Curcumin as an indirect methylation inhibitor modulates the effects of Toxoplasma gondii on genes involved in male fertility.

Toxoplasma gondii is a common protozoan parasite, which infects warm-blooded mammals, including mice and humans, throughout the world. The negative effects of T. gondii infection on the human reproductive system have been documented, especially in females. However, only few studies have examined the effects of T. gondii infection on the male reproductive system. Previous research shows that T. gondii can induce DNA methylation in some gene promoters, which are key regulators of spermatogenesis. Therefore, this study aimed to evaluate the effects of curcumin on the activity of DNA methyltransferases (DNMTs), as well as selected genes, involved in spermatogenesis in spermatogenic cells. In the spermatogenic cells exposed to T. gondii, there was a significant increase in DNMT1 and DNMT3A gene expression and a significant reduction in HSPA1A, MTHR, and DAZL gene expression, compared to the controls. The present results showed that curcumin could regulate changes in T. gondii-mediated gene expression. The effect of T. gondii on DNMT activity was also investigated in this study. A 40 % increase in DNMT activity was observed due to T. gondii infection. However, DNMT activity was restored by treatment with 20 µM curcumin for eight hours. The results revealed that T. gondii increases the NF-κB activity, compared to the control group. The increase in NF-κB activity, induced by T. gondii, was inhibited by curcumin. In conclusion, T. gondii, by increasing DNMT expression and activity, leads to an increase in NF-κB activity in cells. On the other hand, curcumin reduced DNA methylation, induced by T. gondii, owing to its NF-κB-inhibiting properties. Therefore, curcumin, as a hypomethylating agent, can be potentially used to alleviate the negative effects of T. gondii on the male reproductive system.

Genotyping the causative agents of cutaneous leishmaniasis in Ahvaz southwest of Iran.

With sand flies as the main vectors, Leishmania species cause ancient zoonotic diseases called leishmaniasis. Iran is an endemic country regarding cutaneous leishmaniasis. A number of 100 smear slides were collected from cutaneous lesions referred to Ahvaz health centers. The DNA was extracted and ITS1-PCR using LITSR and L5.8S primer pair was performed to detect the genus Leishmania. Then, enzymatic digestion of PCR products was done by HaeIII (species detection), TaqI (strain detection), DpnI and HpaII (mutation assessment). Furthermore, 50 samples were sent for sequencing. Microscopic examination showed amastigote form in all 100 slides. Also, molecular identification confirmed the infection of all cases to Leishmania genus, representing a 350 bp band. HaeIII digestion yielded 150 and 200 bp bands, indicating Leishmania major, while 130 and 200 bp fragments following TaqI digestion suggested A1 strain of the parasite. Moreover, no likely mutations was detected in ITS1 fragment of obtained parasites using DpnI (140 and 200 bp digestion) and HpaII (without digestion). The sequencing result also was consistent with our findings, having 100% homology to A1 strain sequence (AY550178). Leishmania major A1 strain was the predominant species in clinical samples of Ahvaz. Nevertheless, future researches should address the parasite strains in other foci and hosts of epidemiological significance.

In-Silico Identification of the Best Compound Against Leishmania infantum: High Throughput Screening of All FDA Approved Drugs

Objective: Current in-silico research was designed and administered for the screening of 20000 Food and Drug Administration-approved drug compounds with the goal of finding promising drugs against lipophosphoglycan (LPG) and γ-glutamylcysteine synthetase (γ-GCS) of Leishmania infantum. Methods: After the protein sequence of both targets was taken, the 3D structures of protein of interest were predicted and validated. Molecular docking was done among the two putative targets (LPG and γ-GCS) and approved compounds were selected using AutoDock 4.2 program to predict ligand-receptor interactions. Results: After docking experiment was done on 20000 drug compounds, a total number of seven ligands, two for γ-GCS receptor and five for LPG receptor, were assigned as novel, potent anti-leishmanial drugs based on their binding affinity and energy. Of those, five ligands possessed cytotoxic and anti-cancer characteristics and showed good binding capacity to LPG receptor with ΔGbinding up to 8.5 kcal/mol more negative; while two compounds showed good binding capacity to glutamyl receptor with ΔGbinding up to 7.8 kcal/mol more negative. Conclusion: The latest software-based methods are powerful tools for scanning and predicting new peptide templates specific to biological targets in organisms for new drug discovery. However, the use of in vitro and in vivo techniques is a requirement for better evaluation of the potential of projected ligands with the help of in-silico approaches, identifying molecular mechanism of action of the more active compounds is possible. This can help in defining the most likely molecular target, so that the subsequent optimization using in vitro and in vivo techniques can be undertaken.

Molecular diagnosis of potentially human pathogenic Enterocytozoon bieneusi and Encephalitozoon species in exotic birds in Southwestern Iran.

Microsporidia are obligate intracellular parasites that produce spores. The infections caused by these parasites are mostly considered to be opportunistic in immunodeficient patients. Because of the zoonotic nature of microsporidia as well as the increasing prevalence of immunodeficiency diseases, the aim of this study was to evaluate the molecular diagnosis of Enterocytozoon bieneusi (E. bieneusi) and Encephalitozoon spp. in exotic birds in southwestern Iran. Initially, 816 stool specimens were collected and stained by modified trichrome (Weber) staining. The slides were explored using light microscopy. In the next stage, the extracted DNA was amplified using a multiplex/nested PCR method. RFLP with the Mnl1 restriction enzyme was used to differentiate the Encephalitozoon species in the products of the molecular analysis. Out of 816 samples, 138 and 181 cases were found to be positive by the staining and the multiplex/nested-PCR methods, respectively. Of the 181 samples, 103 and 78 samples were positive for E. bieneusi and Encephalitozoon spp., respectively. The Encephalitozoon species were 17 E. cuniculi, 52 E. intestinalis and 9 E. hellem. Of 103 E. bieneusi samples, 57, 39, 2 and 5 cases were detected as genotypes D, M, E and L, respectively. The results showed a relatively high prevalence of microsporidia in exotic birds, and according to the results of the genotyping, these birds can be an important source of microsporidiosis. It is essential that high-risk individuals, including patients with immunodeficiency diseases, receive accurate and valid information about the risk of direct and indirect contact with infected exotic birds.

Seroprevalence and molecular evaluation of toxoplasmosis in children with cancer in Khuzestan province, Southwest of Iran.

Toxoplasma gondii is an intracellular parasite with global distribution. Toxoplasmosis in individuals with normal immune system is usually asymptomatic, but in immunocompromised patients may lead to death if not cured. In this study, the prevalence rate of acute and chronic toxoplasmosis in children with cancer was investigated using serological and molecular methods. Blood samples were taken from 372 children with cancer in Shafa hospital in Ahvaz, southwest of Iran. Anti-T. gondii IgG and IgM antibodies were investigated by ELISA. The presence of Toxoplasma in the blood samples was evaluated by Nested PCR. Among 372 children with cancer, 155 (41.7%) were positive for anti-T. gondii IgG antibodies and 24 (6.4%) were positive for anti-T. gondii IgM antibodies, as well. In IgG avidity test, 34 (22%) had antibodies indicating acute phase and 121 (78%) had antibodies indicating chronic phase. The Nested PCR results were showed T. gondii parasite in 34 (100%) patients among 34 IgG antibody-positive patients with acute infection, among 16 IgG antibody-positive patients with chronic infection, 10 patients were indicative of T. gondii and 6 patients were not indicative of T. gondii. A total of 50 cases, 44 (88%) were T. gondii-positive and 6 (12%) were T. gondii-negative in Nested PCR. This study showed high prevalence of toxoplasmosis in children with cancer. Results of serological techniques (ELISA and IgG avidity) had a higher overlap with Nested PCR in identifying T. gondii of seropositive patients.

Mental Health and Latent Toxoplasmosis: Comparison of Individuals with and without Anti-Toxoplasma Antibodies.

AIM: There is evidence to suggest that the protozoan Toxoplasma gondii affects the mental health of people who are infected with it. The aim of the present study was to examine the relationship between T. gondii and mental health. METHODS: A total of 200 students (87 men and 113 women) of Jundishapur University of Medical Sciences (Ahvaz, Iran) were tested for the presence of anti-Toxoplasma antibodies and completed the General Health Questionnaire (see Appendix 1, available at: http://www.longwoods.com/content/24938) and a demographic form. Data were analyzed using independent samples t-test, chi-square test and Fisher's exact test. RESULTS: Infected women had significantly lower scores in somatic symptoms (p = 0.04), anxiety/insomnia (p = 0.006) and depression (p = 0.04) compared with non-infected women. Difference in social dysfunction was not significant (p > 0.05). There were no significant differences in somatic symptoms, anxiety/insomnia, depression and social dysfunction between infected and non-infected men (all p > 0.05). CONCLUSION: Our findings indicate that latent toxoplasmosis can affect some components of mental health just in women.

Cytotoxic Activity of Holothuria leucospilota Extract against Leishmania infantum In Vitro.

Leishmaniasis is a tropical parasitic infection. The resistance and toxicity issues are the major complications and remain significant consequences related to the treatment of leishmaniasis with the recent and classical drugs. Thus there is an immediate requirement to develop new compounds for the treatment of this protozoan disease. Sea cucumbers or holothurians are potentially presented as the marine sources of antimicrobial and cytotoxic compounds. The aim of this study was investigation of in vitro antileishmanial activity of methanol extract of body wall, coelomic fluid, and cuvierian organs of Holothuria leucospilota obtained from coastal parts of Persian Gulf against Leishmania infantum promastigotes and axenic amastigotes. The colorimetric MTT assay was used to determine L. infantum promastigotes and axenic amastigotes viability at different concentrations of the extracts and drug control (Glucantime®) at time dependent manner and the results are represented as IC50 (50% of inhibitory concentration). Coelomic fluid was the most active extract among the three different extracts of H. leucospilota against L. infantum promastigotes and axenic amastigotes with IC50s of 62.33 µg/mL and 22.4 µg/mL and 73 µg/mL and 46 µg/mL at 48 and 72 hours after treatment, respectively. Cuvierian organs extract showed less toxicity with IC50s more than 1000 µg/mL for both Leishmania infantum axenic amastigotes and promastigotes forms after 48 and 72 hours of exposure. Results acquired from the present study propose that the sea cucumber H. leucospilota may be a provoking source of antileishmanial compounds and could be a lead source in the development of the potent antileishmanial and cytotoxic drugs.

Seroprevalence of toxoplasmosis in diabetic pregnant women in southwestern of Iran.

To investigate anti-Toxoplasma gondii IgG and IgM antibodies in diabetic pregnant women in Ahvaz, southwest of Iran this experiment was performed. In current study the sera of 110 diabetic pregnant women as well as 110 non diabetic pregnant women referred to the hospitals affiliated with the Ahvaz Jundishapur University of Medical Sciences were assessed for anti-T. gondii IgG and IgM antibodies by ELISA and IFA methods. The ELISA assessments showed that 47 (42.7 %) and 3 (2.7 %) of diabetic women were positive for IgG and IgM antibodies, respectively. However, in the control group, 24 individuals (21.81 %) were positive for IgG antibody but no detection for IgM antibody. According to IFA method, 46 (41.8 %) and 3 (2.7 %) of diabetic women were positive for IgG and IgM antibodies, respectively, while in control group, 21 individuals (19.09 %) were positive for IgG antibody. In this method, IgM antibody was negative for all samples of control group (0 %). In both methods, the values obtained in the case group were significantly higher than those in the control group (p < 0.05). Prevalence of anti-Toxoplasma IgG and IgM antibodies in diabetic pregnant women was higher than that in non-diabetic pregnant women. It seems that screening tests for seeking patients and teaching the transmission routes should be considered as prenatal cares for diabetic women.

Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes.

BACKGROUND: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro. METHODS: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC50) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively. RESULTS: The IC50 value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 µg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed. CONCLUSION: The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.