فهرست مطالب abdol hossein dalimi asl

Leishmaniasis is a protozoan zoonosis that causes problems in public health. The use of pentavalent antimony compounds as chemical drugs in the treatment of cutaneous leishmaniasis is associated with various limitations and side effects. Today, plant extracts and their derivations are the promising sources in herbal medicine. The purpose of this study was to evaluate the effectiveness of oil extract Pistacia atlantica  Kurdistan province, Iran, on promastigotes of standard strain Leishmania major in vitro and in vivo. n this study, 10, 15, 25, and 50% concentrations of plant-based ointment were added to the lesions caused by promastigotes of Leishmania major. The effectiveness of treatment with different concentrations of pistacia extract on induced lesions at the base of the tail of Balb/c mice cultured with promastigotes of Leishmania major was checked in terms of disease progression or improvement and parasite load. The effect of 50% pistacia extract has the greatest effect on reducing the lesions diameter of Leishmania promastigotes, the results showed that there is a significant difference between the number of Leishmania major promastigotes in different concentrations of the extract and exposure time. With respect to the calculated P-value for time variable (P < 0.05), there was statistically remarkable difference among different time periods, so the mean of the measured lesion was different regarding 6 weeks of experiment, and there observed a statistically significant difference in terms of the time of lesion measurement. Additionally, the prediction, isolation, purification, and application of the effective compounds found in the Pistacia atlantica extract is highly suggested for future studies.

Acanthamoeba is an opportunistic pathogen that may cause fatal granulomatous encephalitis and ocular keratitis in humans and animals. Increasing number of contact lens wearers can lead to increased frequency of amoebic keratitis and due to lack of effective drugs treatment of this disease is difficult. This study aimed to investigate the effect of ethanolic extract of Chaerophyllum macropodum on Acanthamoeba genotype T4.

In this experimental study, samples taken from the patients with keratitis were cultured on 1.5% non-nutrient agar medium. Different concentrations of the ethanolic extract (1.25, 2.5, 5, 10 mg/ml) were tested three times (24, 48, and 72 h) on trophozoites and cysts of Acanthamoeba in vitro. The number of live and dead parasites were counted by using trypan blue staining and neobar lam. Also, the percentage of cysts apoptosis was assessed by flow cytometry.

In the presence of 10 mg/ml ethanolic extract in the culture medium, the percentages of live trophozoites after 24, 48, and 72 h were 53.6, 15.11, and 0 percent, respectively. In the case of cysts, after 24, 48, and 72 h, 65.31, 43.31, and 0 percent of the cysts were alive, respectively. The results of flow cytometry showed that the apoptosis rate was 0.09% in the sample treated with extract after 72 h.

Ethanolic extract of Chirophyllum macropodium can be a promising candidate for the development of anti-Acanthamoeba drugs due to its high toxic effects on the cysts. However, since the crude extract of Chaerophyllum macropodum was used in this study, further investigations are needed to find the effective fractions of the plant and determine their mechanisms of action. It can also be tested in vivo or even against other parasites

Amoebae of the genus Acanthamoeba are unicellular amphizoic opportunistic pathogens that may cause fatal granulomatous encephalitis, eye keratitis, amebic pneumonitis and skin nodules as well as abscesses in humans and animals. Acanthamoeba keratitis is caused by trauma to the eye, contaminated cleaning solutions and the use of contact lenses. The aim of the present study was to identify the genotypes of Acanthamoeba in all patients with a clinical diagnosis of Acanthamoeba keratitis referring to eye clinic in Tehran using polymerase chain reaction (PCR).

In this study, samples were collected from 35 patients who had referred to the eye clinic and were cultured on 1.5% non-nutrient agar. DNA was extracted, and then PCR amplification was performed using genus specific primers. Sequencing analysis and basic local alignment search tool search were conducted to determine the genotypes. Phylogenetic tree was generated using maximum likely algorithm in phylogenetic program MEGA version 6.

Eight cases were positive for Acanthamoeba using genus specific primer pairs. All specimens were reported as genotype T4.

Determination of genotypes showed all isolates belonging to genotype T4; this abundance may be due to its higher prevalence in the environment or its greater virulence. However, further analysis of clinical and environmental samples is necessary to clarify this property.

Gyrodactylus, a member of Platyhelminthes, is one of the most common external parasites on freshwater and marine fish. This parasite mostly appears on skin and fins but rarely on gills of fish. Gyrodactylus can cause disease and mortality in wild and domestic populations of fish. In this study Gyrodactylus specimens removed by wet mounts of skin and fins of Cyprinus carpio, Hypophthalmichthys molitrix and Hypophthalmichthys nobilis in Guilan Province fish ponds and analyzed by a light microscope. The morphometrical identification of Gyrodactylus specimens was performed using the measurements and drawing of opisthaptoral hard parts like as anchor, marginal hook, ventral bar and dorsal bar. The molecular species description was based on polymerase chain reaction (PCR) of a partial sequence of the 5.8S region of ribosomal RNA, and a partial sequence of the internal transcribed spacer 2 (ITS2) of ribosomal RNA. The nucleotide sequences of the PCR products were compared with other sequences registered in GenBank. Based on the morphometric analysis and sequencing, the Gyrodactylus specimens were identified as Gyrodactylus sprostonae. The abundance of this parasite in investigated fish ponds of Guilan Province were 21.42, 6.17, 21.95 and zero for C. carpio, H. molitrix, H. nobilis and Ctenopharyngodon idella respectively.

Toxoplasmosis is a common infection all around the world. During pregnancy; it may lead to congenital disorders or abortion in human and animals. Severe damage of toxoplasmosis indicates to require effective vaccine. One of dense granules antigen is GRA4 that secrete from tachyzoite and bradyzoite. GRA4 genome is unique without intron and is one of the major immunogenic proteins from Toxoplasma gondii. We confirmed the cloning of GRA4 gene into pcDNA3 by restriction enzyme and PCR of GRA4 gene with pcGRA4 plasmids as template. Then with using calcium- phosphate method we transfected the pcGRA4 into CHO (Chinesehamster ovary) cells. The yielded protein was separated by SDS-PAGE and moved by electroblotting to nitrocellulose paper. Result of SDS-PAGE analysis showed the appearance of band approximately 42 kDa which was absent in the negative control, that was able to identify toxoplasmosis antibody IgM+ serum in western blot analysis. pcGRA4 plasmid is able to synthesis of antigenic protein in CHO cells. The ability of pcGRA4 for induction of protective immune response against toxoplasmosis will be evaluated in mouse model.

Many plant extracts have antiparasitic properties and even some of them are effective for in vivo treatment. One of the medicinal plants is Peganum harmala, which can grow from 30 to 100 cm. The aim of the present study was to determine the effect of aqueous extract of Peganum harmala on trophozoites and cysts of Acanthamoeba in vitro.
In this experimental study, a sample from a patient with amoebic keratitis, was cultured with Escherichia coli bacterium in non-nutrient agar medium (NNA) 1.5%. Then, the aqueous extract of Peganum harmala was prepared. The extract was affected on trophozoites and cysts of Acanthamoeba at three times (24, 48, and 72h) in various concentrations (1.25, 2.5, 5, and 10mg/ml). The percentage of live parasites, was evaluated using homocytometer lamel, MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], and flow cytometry method. The data were analyzed using one-way ANOVA, one sample Kolmogorov-Smirnov test, and repeated measures test The significance level was considered to be p The extract showed dose- and time-dependent activity on trophozoites and cysts. In the presence of 10 mg/ml aqueous extract in culture medium, 32.47% of the trophozoites and 50.15% of the cysts, were alive after 72 hours.
Our study indicated that the aqueous extract of the Peganum harmala, has anti-Acanthamoeba activity. Therefore, it is recommended that the effect of Peganum harmala extract be studied in animal models.

The objective of this study was to investigate HLA-DRB1*and DQB1* allelic polymorphisms in Iranian patients with hydatidose. This is the first survey dealing with the correlation between HLA-DRB1* and DQB1* al­leles and cystic echinococcosis in Iranian patients. The study was carried out on 56 patients with confirmed cystic echino­coccosis and 30 apparently healthy individuals living in Arak- Markazi Province by HLA-DRB1 and DQB1 typing through PCR-SSP method. The first step was to identify the patients and blood sampling. DNA was prepared from whole blood and PCR-SSP with 31 primer mixes for per sample was used. PCR reaction mixtures were loaded in agarose gels and bands were observed under UV illumination and gel document after electrophoresis. Analysis of re­sults was carried out with specific softwares and frequency and interpretation tables for calculation of P-value in χ2 test were provided via Fisher΄s exact test. Significant samples were analyzed by logistic regression and odds-ratios were calculated. A statistically significant positive association was found between HLA-DQB1*03 and the resistance to cystic echinococcosis (P<0.02) (odds-ratio=2.87). Immunogenetic susceptibility to unilocular hydatidose varies accord­ing to the HLA antigens in Arak, Markazi Province, and DQB1*03 mole­cules are associated with the level of immune response to parasite antigens.

Application of quantitative real time PCR has evolved as a sensitive, specific, and rapid method for the detection of Toxoplasma gondii (T. gondii). The present study aimed to evaluate the efficacy of real time PCR method, using B1 gene, for the diagnosis of toxoplasmosis in the experimentally infected rats. Parasites were cultured in peritoneal cavity of mice and then the DNA was extracted in tachyzoite stage. The B1 gene of T. gondii was amplified by PCR and detected by real time PCR method based on the molecular beacon probe. Finally, real time PCR was evaluated for the quantization of T. gondii in the blood of the experimentally infected rats. The B1 gene of T. gondii which was successfully amplified by PCR yielded an amplicon with an approximate length of 116 bp. Using this gene was evaluated highly appropriate for the quantization of T. gondii by real time PCR method. Application of real time PCR method is shown to be highly efficient in terms of sensitivity and rapidity for the detection of B1 gene as well as the quantization of T. gondii in blood of rat.

ÏL22 is the family member of ÏL10 that is created by Th17، Th22، NK cells and recently the role of Ïl22 in protection and inherent defense mechanisms، in the control of bacterial and viral infection، homeostasis and repair tissue has been demonstrated. So in this research the effect of Ïl22 treatment on lesion originated from L-major in BÂLB/c mice has been survived.

24 female BÂLB/c mice for 8 weeks old at least 2x10 6 Promastigotes of L-Major Ïraninan standard strain MR HÔ/ÏR/75/ËR in stationary phase through 100 micro liter subcutaneous challenged and in three of eight groups of mice has been injected by 5ng/ml and 10ng/ml (ÏM) with recombinant ÏL22. Ïmmunity cellular and homoral with assessment cytokines ÏL 4

ÏFN-ɤ، cultures spleen lymphocyte cells and survey of ÏgG2a، ÏgG Total with Ëlisa method and MTT method and clinical study measuring the wound healing and lifespan of mice and death registration was done. Çonclusion: The study conclusion indicated that the growth of ulcer especially has been reduced in ÏL-22 groups and the greatest effect in ÏL-22 (5ng) group. ÏL-22 (5ng) has caused the increased production of، ÏFN- ɤ and the reduced ÏL4. The LSD test has shown that، ÏFN- ɤ، ÏGg Total with P<0. 05 significant in ÏL22-5ng groups rather than other groups and ÏL4 with P<0. 05 non significant in ÏL22-5ng but significant with other groups. ÏGg2a in ÏL22-5ng with P<0. 05 is not significant with ÏL22-10ng groups. but significant with other groups. MTT ÏL22-5ng is significant with other groups. We can infer that increasing ÏFN-ɤ and reducing ÏL4 by ÏL22-5ng has caused the protective immunity in L-major، so if it is used with the anti leishmania drug combined may have effective results in treatment.

An entomological survey was carried out for Leishmania vector incrimination of sand flies in northwestern Iran. Among other specimens, 358 sand flies belong to the Sergentomyia Genus were tested for leptomonad infection using semi-nested PCR method as well as sequence anlalysis of ITS-rDNA fragment. Results of semi-nested PCR against kietoplast DNA showed reptile leptomonad infection in two specimens of S.dentata. The ITS2 sequence analysis of the specimens revealed 76% identity with those of Leishmania (sauroleishmania) adleri of Genbank. However, further studies need to clarify the species identity of the leptomonads. Interestingly, blood meal analysis of the sand flies determined an S.sintoni specimen with mammalian hemoglobin. This reptile related sauroleishmania parasites lacks the Lipophosphoglican (LPG) necessary for entrance to human phagocytes cells, and hence are not human pathogen. However, the GlycoInositoPhosphoLipid (GIPL) molecules of this parasite reacts with sera of kala-azar patients and may cause false positive scores in sero-epidemiological surveys for kala-azar. Sauroleishmania can be transmitted to human by infected bite of some Sergentomyia subgenera that show intermediate

Toxoplasmosis is a protozoal infection caused by Toxoplasma gondii. Toxoplasmosis produce severe damage in patients who are immunosuppresed. In those who are immunosupressed, latent infection can be reactivated resulting in acute disseminating disease. Betamethasone is a synthetic glycocorticoid, used as an anti-inflamatory and immunosuppressant in a wide variety of disorders.The aim of this study was evaluation of betamethasone as an immunosuppressor drug on infected cells by Toxoplasma gondii. In this study, at first HeLa cells were grown in 24 well culture plates in culture medium. When confluent monolayer was obtained, we compared 6 groups to evaluate the effect of betamethasone as a corticosteroid drug (two concentrations 4 and 40μg/ml) and the effect of IFN-γ (100 IU/ml) on growth, replication and Nitric Oxide (NO) production. The results showed, that high number of plaques were seen in group with 40 μg/ml of betamethasone and the lowest number of plaques were seen in group with 100 IU of IFN-γ. The difference between plaque number in control and groups treated with IFN-γ and betamethasone was significant (P<0.05). The groups with betamethasone or IFN-γ without tachyzoites did not show any effect on cell structures. Replication rates in the wells treated with IFN-γ were decreased significantly 72h post inoculation in comparison with control group (P<0.05). There was no significant difference among different groups in NO production. The results indicated that betamethasone increase the invasion of tachyzoites to host cells in vitro.

Toxoplasma gondii is a coccidian protozoon which forms tissue cyst in different organs of infected intermediate host. The present study was conducted to demonstrate the active form of parasite in different tissues of rat, which was experimentally infected with RH strain of Toxoplasma gondii using bioassay method inmice. In the experimental assay, 75 rats and 500 mice were used. The rats were infected experimentally with 40-50 thousands of tachyzoites intrapritoneally. Three rats were killed every 24 h then with up to three days 3 days interval for 60 days and their different organs were searched biologically for presence of the parasite. In this regard, the organs were squeezed with mortar in normal saline, diluted and following adding streptomycin and penicillin injected intrapritoneally in three outbreed mice each. Then the mice were followed up for 10 days. Toxoplasma cyst was found in brain, liver and spleen of the rats within 5 to 9 post infection. The parasite was appeared in heart and muscles within 9-13 days after infection but not found in the kidney. The parasite was disappeared in spleen and liver following12 and 28 days of infection respectively but remain active in the brain, heart and muscles within 60 days of the study.Implication: Toxoplasma gondii could remain active in spleen and liver of infected rat for a short time. Meanwhile brain and muscles (heart and skeletal muscles) of the infected animal can keep the parasite for a long time. Kidney is not appropriate tissue for Toxoplasma gondii localization.

Considering the importance of hydatid cyst and its pathophysiological effects on the human body, we attempt to propose a method for killing of hydatid cyst protoscoleces in vitro condition using low voltage direct electrical current.

After collecting hydatid cyst from infected organs of slaughtered animals, their protoscoleces were cultured in four separated media of hydatid consisting of uid, RPMI, normal saline and Tris buffer. The protoscoleces with the same media were transferred to an electrolysis device, and then various sets of electric current density were applied. For measuring the survival rate of protoscolcces, the same cells movement as well as Eosin staining were used as standard techniques.

Our findings indicated that the survival rate of protoscoleces in hydatid fluid was dependent on the electric current density and the time of the applied current. In this regard, the highest survival rate of 86.3% was observed when electric current density of 42.96 mA/cm2 was applied for one minute; while the survival rate of 0% was observed when an electric current density of 63.5 mA/cm2 was applied for one minute. In the RPMI, normal saline and Tris buffer media, similar results were obtained.

Low voltage of direct current can be used in destroying of protoscoleces during surgery on different body organs without any injury to host tissue and any relapse of the infection.