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  Introduction and objective: Dicrocoelium dendriticum is a worldwide spread parasite of liver, bile ducts and gallbladder of especially ruminants and humans as well. Identification of specific antigens is useful for early diagnosis of the infection. The goal of this study was the isolation and identification of excretory-secretory and somatic antigens from D. dendriticum by sodium dodecyl sulphate (SDS)-PAGE and evaluation of humoral immune response against these antigens.

  Methods: The parasites were collected and washed by phosphate buffered saline (PBS) and supplemented by antibiotic for several times. For preparing somatic antigens, parasites were sonicated and centrifuged prior to collect supernatant. For preparing excretory-secretory antigens the viable parasites were transferred to the sterile medium. The samples were centrifuged and supernatants were collected. The sera of infected sheep with different infection degrees were collected too. Somatic and excretory-secretory proteins were isolated with SDS PAGE and stained with coomassie blue. Immunogenicity properties of the resulting proteins were determined using western blot analysis.

  Results: The total extract of somatic antigens analyzed by SDS-PAGE revealed 21 proteins. In mild infection, bands of 130 KDa were immune dominant. In moderate infections 48, 80 and 130 KDa and in heavy infections 48, 60, 80, 130 KDa were detected as immune dominant bands. In excretory- secretory antigens seven bands of protein were detected. In mild infection 130 KDa, in moderate infection 100, 120 and 130 KDa and in heavy infection 45, 80, 85, 100, 120 and 130 KDa were immune dominant bands.

  Conclusion: Probably the most immunogenic protein band during different degrees of infection was 130KDa that can be used for vaccination and inducing immunity.

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  Background & Objectives: Regarding to high incidence of dysmenorrhea and influence on daily activities and fewer side effects of herbal medicines than chemical drugs, the aim of this study was to compare the effect of mefenamic acid and matricaria chamomilla (MC) on primary dysmenorrhea.

  Methods: This triple-blind randomized clinical trial study was done on 90 female students residents in dormitories of Kashan University of Medical Sciences in 2012. The subjects were categorized into two groups randomly. Mefenamic acid capsules (250 mg, every 8 hours) were given to the first group from 48 hours before menstruation until 24 hours after it. The second group received MC capsules made in Barij Essence Factory of Kashan (250 mg, every 8 hours). Severity of dysmenorrhea was measured by McGill ruler. Finally, the data were analyzed by SPSS. The chi-squire, fisher and paired t-test were used. The p-value of less than 0.05 was considered as statistically significant difference.

  Results: The result of this study indicated that both chamomilla and mefenamic acid can reduce the severity of pain and hemorrhage (p<0.05) but there was no significant difference between two groups (p>0.05).

  Conclusion: This study showed that matricaria chamomilla is effective in decreasing the severity of primary dysmenorrhea and reducing hemorrhage as well as mefenamic acid.

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  Background & objectives: The leishmaniases are considered among the major infectious diseases affecting public health in several regions. There are many chemical agents which are effective in treatment of visceral leishmaniasis. But, overall treatment of visceral leishmaniasis is often difficult. Thus, identification of new chemotherapeutic agents is important for treatment of disease. Since targeting of the ergosterol synthesis pathway of Leishmania may be useful therapeutically, the aim of this study was to investigate the effect of alone or in combination of amiodarone and ketoconazole on Leishmania infantum.

  Methods : To obtain logarithmic promastigotes of L. infantum, the parasites were cultured in BHI medium with FCS 10% together with antibiotics of penicillin and streptomycin and incubated at 24° C. Amastigote forms were obtained in BHI medium supplemented with 20% FCS at pH of 5.5 which incubated in 37° C. L.infantum susceptibility to amiodarone and ketoconazole was evaluated by proliferation of parasites in the absence or presence of these drugs with MTT assay. For evaluation of antiproliferative synergism against promastigotes and axenic amastigotes, fractional inhibitory concentrations (FIC) were calculated. An isobologram curve was constructed too.

  Results: Amiodarone produced a marked reduction in the viability of L.infantum promastigotes and axenic amastigotes. On the other hand ketoconazole induced a dose dependent effect on the parasites proliferation for promastigotes and axenic amastigotes. When the drugs were used in combination, the results indicated clear synergistic as shown by a concave isobologram and FIC value.

  Conclusion: The present study represents the evidence that the combination of amiodarone plus ketoconazole acts synergistically in controlling L. infantume in vitro. It is possible that amiodarone could be used in combination with ketoconazole to combat infection at low doses, thus reducing its side effects such as cardiotoxicity, thyroid dysfunction and pulmonary fibrosis.

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Background & objectives: One of the most important zoonotic parasitic diseases, hydatidosis, is caused by the larval stage of Echinococcous granulosus. Investigations have shown that plants secondary metabolites, such as essential oils have anti parasitic properties. Based on previous reports on antiparasitic properties of Lepidium sativum, in this study we investigated the scolicidal effects of the essential oil (EO) extracted from this plant.

Methods: Lepidium EO was obtained by hydrodistillation method. Gas chromatography-mass spectrometry (GC-MS) was employed to determine the chemical composition of the EO. Protoscolices were exposed to various concentrations of EO (1, 3, 5, 10 and 15 mg/ml) for 10, 20, 30 and 60 min. Viability of protoscolices was confirmed by 0.1% eosin staining.

Results: A total of 19 compounds representing 95.5% of the total oil, were identified. &alpha;-Thujene (88.86%), Myrcene (2.9%) and P-cymene (1.67%) were found to be the major EO constituents. Based on the results, protoscolices mortality rates at 1, 3 and 5 mg/ml of EO didn’t have a significant relationship with the control group. While, the difference in mortality rate at a concentration of 10 mg/ml of EO in 30 and 60 min was significant. Also, the concentration of 15 mg/ml of EO at all times of incubation had significantly higher protoscolicidal effect. In the present study there was a significant relation between the amount of protoscolicidal activity of different EO concentrations and different incubation times. In other words mortality rates enhanced with increasing concentrations and incubation times.

Conclusion: The results of present study revealed that the EO of Lepidium is rich in &alpha;-Thujene and has a high scolicidal power. This plant may be used as a natural scolicidal agent

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Background & objectives: The adjacent of residential buildings in the countryside with livestock causes external parasites to be transferred easily and feed on the human hosts. Due to fleas haematophagus nature they are able to transfer pathogens from animal to animal or animal to human and thus they are considered as zoonotic pathogens. Therefore, identification of fleas is necessary.

Methods: In the present study 30 infested people with biting signs and 800 sheep and goats were investigated. About 50 fleas from infested people and 160 from animals were collected. Samples were cleared with KOH and recognized based on proper identification keys.

Results: Based on the results it seems that sheep and goats were infested with Ctenocephalides canis and Pulex irritans. Out of the 160 studied fleas from sheep and goats 118 (73.7%) were identified as C. canis and 42 (26.3%) as P. irritans. Out of 50 collected fleas from infested people 43 (86%) were identified as C. canis and 7 (14%) as P. irritans.

Conclusion: The present report is the first report of man infestation with canine fleas or C. canis. According to climate condition and employment of most of villagers to traditional animal husbandry, it seems that there is a proper condition for external parasites (such as fleas) growth and proliferation. Therefore, studies based on infestation identification and report can be considered for control strategic programs.

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Background & objectives: Polyamines such as putrescine, spermidine, and spermine are ubiquitous in all eukaryotic cells and play an essential role in cell division and differentiation. One way of polyamine biosynthesis is done by ornithine decarboxylase (ODC) which catalyzes the transformation of ornithine to putrescine. The aim of the present study was to evaluate the level of putrescine, spermidine and spermine in protoscolices, hydatid fluid and germinal layer and also to evaluate ODC activity.

Methods: In the present study putrescine, spermidine and spermine levels were investigated in germinal layers, hydatid fluids and protoscolices. To evaluate the activity of ODC, protoscolices were incubated with ornithine and changes in polyamines level were assayed. The samples were homogenized and liquid chromatography (HPLC) was used for polyamines measurement.

Results: Based on the results, putrescine was the lowest polyamine and since its level was not increased in protoscolices incubated with ornithine, ODC activity was not detected. Spermidine was the highest polyamine and the results showed that germinal layer contained the highest level of polyamines.

Conclusion: Overall, the results showed that ODC activity was not detected in hydatid cyst and level of polyamines in germinal layers  which contained rapidly proliferating cells was higher than other parts.

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Background & objectives: Toxoplasma gondii is a zoonotic obligate intracellular protozoan parasite that infects all warm-blooded animals as well as human worldwide. Determining the parasite genotype in intermediate hosts  is crucial in  evaluating the role of these types in human infections as wll as in prevention programs. Therefore, this study aimed to identify and detect the genotypes of Toxoplasma gondii in aborted fetuses of ewes in Lorestan province.
Methods: Identification of the parasite was performed  on the brain and liver tissues of 142 aborted fetuses using  a conventional PCR based on amplification of highly repetitive 529 bp region of the parasite genome. Genotyping of positive samples, which were isolated from the brain and liver, was performed by PCR-RFLP based on SAG2, SAG3 and GRA6 molecular markers.
Results: From a total of 142 samples  obtained from  brain  and fetus, 10 cases (7%) were determined as positive samples based on conventional PCR. The precence of parasite DNA was also confirmed in the liver of  3 positive samples. Evaluation of RFLP pattern of amplified SAG2, SAG3 and GRA6 genes showed the presence of various types of parasites, incuding type I in 3 samples, type II in 2 samples and atypical type in 5 samples.
Conclusion: Isolation of types I, II and atypical type of T. gondii from ewes in  Lorestan province suggests the need for greater attention to parasite transmission from livestock to human, particularly in pregnant women and people with weakened immune system.

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Background & Objectives: Cold Plasma is an emerging non-thermal, chemical-free, environmentally friendly disinfection technology. Plasma-activated water has received considerable attention from researchers in recent years. Despite extensive studies on the antibacterial effects of plasma-activated water, its anti-eukaryotic effects have not been identified. In humans, Acanthamoeba causes granulomatous encephalitis, skin ulcers, and Acanthamoeba keratitis. Considering the health importance of Acanthamoeba, this study investigated the anti-amoeba  effect of plasma-activated water on trophozoites and cysts of Acanthamoeba castellanii.
Methods: In this study, plasma-activated water prepared by the cold atmospheric plasma method.Physicochemical properties of produced water were evaluated by measuring pH, hydrogen peroxide, nitrite, and nitrate. To assess the effect of plasma-activated water on A. castellanii, trophozoites and cysts were exposed to plasma-activated water for 0.5, 1, 2, 3, 4, and 5 hours. Three replicates were examined each time. At the mentioned times, cell viability was calculated by trypan-blue staining and counting on a hemocytometer, and the results were statistically analyzed.
Results: Based on the physicochemical results, the mean pH of plasma-activated water in this study was about 3.4, and the amount of hydrogen peroxide, nitrate, and nitrite were 102, 737, and 36.94 μM, respectively. The present study showed that plasma-activated water killed A. castellanii trophozoites after three hours of exposure and A. castellanii cysts after four hours of exposure. On the other hand, some trophozoites gradually became cysts after exposure to plasma-activated water. These cysts became more resistant to plasma-activated water and inactivated after five hours of exposure.
Conclusion: In this study, for the first time, the effect of plasma-activated water on A. castellanii was investigated. The results of the present study showed that plasma-activated water is able to inactivate A. castellanii trophozoites and cysts. Therefore, plasma-activated water can be used to disinfect and inactivate A. castellanii.

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Background & objectives: Endurance activity affects muscles through changes in hormone- secretion and related receptors. The aim of this study was to evaluate the effect of endurance training on Thrap1 gene expression in cardiac tissue and fast and slow twitch skeletal muscles in male Wistar rats.
Methods: The subjects of this experimental study were 14 male rats with a mean and standard deviation of 234±34g, all of which were kept in natural conditions (free access to water and food, cycle of darkness and light, suitable temperature and humidity). They were randomly divided into two groups of control (n=7) and experimental (n=7). The experimental group had endurance activities 6 sessions per week at the speed of 30 meters per minute for 14 weeks. 48 hours after the last training session, they were anesthetized and dissected under sterile conditions, and Real- time RT-PCR method was employed to determine the gene expression. Finally,a t-test was used to evaluate the data.
Results: The results of this study showed that the expression of the Thrap1 gene in the soleus muscle (p=0.001) and heart (p=0.001) of experimental rats increased significantly, while there was not a significant change in the expression of the Thrap1 gene in fast twitch muscles (p=0.508) due to endurance activity.
Conclusions: It seems the expression of the Thrap1 gene in slow twitch muscles is more affected than fast twitch muscles by endurance activity.
 

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