Effect of Crocus sativus Stigma (saffron) alone or in combination with chloroquine on chloroquine sensitive strain of Plasmodium berghei in mice

Introduction: In malaria treatment protocols, treatment failure or drug resistance of synthesized drugs like alkaloids related to quinine, and aminoquinolines are the main problems now. Therefore, discovering efficient drugs or combination therapy of blood schizonticidal drugs with different mechanisms or different targets in the parasite is a crucial effort to solve this problem. In this study, the effectiveness of Crocus sativus Stigma (saffron) individually and in combination with chloroquine, was considered against chloroquine–sensitive strain of Plasmodium berghei.Methods: At the first stage, using 4 day suppressive Peter’s test in mice, ED50 and survival times of saffron methanol extract, and its aqueous and ethyl acetate fractions and chloroquine on P. berghei were calculated. Then, based on the toxicity and survival time results, combination therapy was conducted with the best saffron fraction and chloroquine against the parasite.Results: The saffron extract, aqueous and ethyl acetate fractions resulted in suppression of parasitemia with ED50 values of 587.0 ± 78.7, 323.7 ± 37.2, and 508.7 ± 35.6 mg/kg, respectively. Combination of ethyl acetate fraction with chloroquine, potentiated the antimalarial property and the survived percent of the treated mice on days 7, 14, and 28 significantly more than chloroquine or ethyl acetate fraction alone.Conclusion: Saffron and its fractions individually can be effective in reducing the parasitemia in mice. The outcome of combination of ethyl acetate fraction with chloroquine on the mice showed synergistic effect on the chloroquine–sensitive strain of parasite

The Role of Liposomal CpG ODN on the Course of L. major Infection in BALB/C Mice

"nBackground: Historically, leishmanization is the most effective protective measure against Cutaneous Leishmaniasis (CL), CL lesion induced by leishmanization sometimes takes a long time to heal. Ma­nipulation of leishmanization inoculums needed to induce a mild and acceptable CL lesion. The aim of this study was to explore if liposomal form of CpG ODN (Cytosin phosphate Guanin Oligodeoxynu­cleotides) mixed with Leishmania major   would induce a milder lesion size in Balb/c mice."nMethods: This study was performed in Biotechnology Research Center, Mashhad, and Center for Re­search and Training in Skin Diseases and Leprosy, Tehran, Iran during 2008-2009.  mice were subcutaneously (SC) inoculated with L. major mixed with liposomal form of CpG ODN, or L. major plus free CpG ODN, or L. major mixed with empty liposomes or L. major in PBS. The lesion onset and the size of lesion were recorded; the death rate was also monitored. "nResult: Footpad thickness was significantly (P<0.01) smaller, death rate was also significantly (P<0.05) lower in the mice received L. major mixed with liposomal CpG ODN or free CpG ODN than control groups received L. major in PBS or L. major plus liposomes, also mice which received L. ma­jor mixed with CpG ODN in soluble form showed a significantly (P < 0.001) smaller lesion size than control groups."nConclusion: CpG ODN seems to be an appropriate immunopotentiator mixed with Leishmania stabi­late in leishmanization

Effect of Toxocara canis and Toxascaris leonina egg antigens on induction of eosinophilia in animal model

زمینه و هدف: توکسوکاراها و توکساسکاریس انگل روده سگ و گربه می باشند که لارو آن ها باعث ایجاد ائوزینوفیلی در انسان می شود. در این تحقیق به صورت اختصاری تأثیر آنتی ژن های تخم برخی از این انگل ها بر ایجاد ائوزینوفیلی بررسی شده است. روش بررسی: در این مطالعه تجربی تعداد 54 سر موش سوری در شش گروه قرار گرفتند. به گروه های مورد، آنتی ژن تخم توکسوکارا کانیس یا توکساسکاریس لئونینا بدون ادجونت از راه داخل صفاقی و یا همراه با ادجوانت فروندز از راه زیر جلدی تزریق شد. گروه های شاهد هیچ تزریقی دریافت نکردند. هر تزریق سه بار با فاصله زمانی دو هفته تکرار و بعد از هر تزریق، شمارش گلبول های سفید از نمونه های خونی انجام شد. یافته ها: اختلافی بین میانگین گلبول های سفید شامل لنفوسیت ها، ائوزینوفیل ها، نوتروفیل ها، مونوسیت ها و بازوفیل ها در گروه های مورد در مقایسه با گروه های شاهد مشاهده نگردید. نتیجه گیری: بر خلاف لاروها، آنتی ژن های انگل های مورد مطالعه باعث ائوزینوفیلی در موش ها نشدند؛ با این حال تحقیقات بیشتری در این خصوص توصیه می گردد

Molecular and microscopy detection of Pneumocystis jirovecii in hospitalized patients during the COVID-19 pandemic

IntroductionEarly detection of Pneumocystis jirovecii as an opportunistic pathogen that may endanger predisposed persons, including COVID-19 patients, may help to choose the optimal management.MethodsIn this study, 585, including 530 COVID-19 patients, with clinical and radiological evidence of respiratory diseases, were investigated for P. jirovecii screening. Clinical specimens were examined by direct microscopy and PCR, and randomly selected positive PCR products were confirmed through DNA sequence analysis.ResultsThirty-one (5.3%) samples were positive in P. jirovecii-specific nested-PCR, while by direct microscopic tests, Pneumocystis was observed in 22 (3.76%) samples. Males (61.7%) and patients over 50 years old (75.6%) were more commonly affected than others, and malaise and fatigue (84%), and wheezing (75%) were the most common symptoms, followed by fever (40.48%) and dyspnea (39.51%). Among the Pneumocystis-positive patients, three cases had coinfection with Aspergillus fumigatus, A. flavus, and A. niger (each n = 1), as documented by direct microscopy, culture, and species identification by PCR-sequencing.ConclusionPneumocystis pneumonia is still a diagnostic challenge; therefore, additional large-scale studies are needed to clarify the epidemiology of the disease in immunocompromised or COVID-19 patients

Echinococcus granulosus : epidemiology and state-of-the-art of diagnostics in animals

Diagnosis and detection of Echinococcus granulosus (sensu lato) infection in animals is a prerequisite for epidemiological studies and surveillance of echinococcosis in endemic, re-emergent or emergent transmission zones. Advances in diagnostic approaches for definitive hosts and livestock, however, have not progressed equally over the last 20 years. Development of laboratory based diagnostics for canids using coproantigen ELISA and also coproPCR, have had a huge impact on epidemiological studies and more recently on surveillance during hydatid control programmes. In contrast, diagnosis of cystic echinococcosis (CE) in livestock still relies largely on conventional post-mortem inspection, despite a relatively low diagnostic sensitivity especially in early infections, as current serodiagnostics do not provide a sufficiently specific and sensitive practical pre-mortem alternative. As a result, testing of dog faecal samples by coproantigen ELISA, often combined with mass ultrasound screening programmes for human CE, has been the preferred approach for monitoring and surveillance in resource-poor endemic areas and during control schemes. In this article we review the current options and approaches for diagnosis of E. granulosus infection in definitive and animal intermediate hosts (including applications in non-domesticated species) and make conclusions and recommendations for further improvements in diagnosis for use in epidemiological studies and surveillance schemes

DEPARTMENT OF INTERNAL MEDICINE, SCHOOL OF HEALTH, ISFAHAN UNIVERSITY OF MEDICAL SCIENCES AND HEALTH SERVICES, ISFAHAN, IRAN

Introduction: Cryptosporidium is a parasite from coccidian order and one of the most common causes of diarrhea in the world, which can lead to a severe and prolonged disease in immunodeficient patients. There is a discrepancy regarding the prevalence rate of this parastie in different studies in our country patients on dialysis are usually prone to infectious disease especially those caused by opportunistic organisms. We studied the prevalence rate of the cryptosporidium infection in a group of patients on dialysis who were considered to have acquired immunodeficiency.&#13; Methods: This is a descriptive analytic study which included 104 dialyzed patients on dialysis in Al-zahra, shariati and Ali-asghar hospital, Isfahan, Iran. Between January 2001 and October 2001. Cases were chosen according to convenience sampling standards. They were all tested for cryptosporidium infection. Test results in the case group were then compared with the rate of cryptosporidium infection. In the control group which included 91 healthy household family members of the patients in the case group and 140 healthy people from the society. At least two samples were collected from every subject on two different occasions. To detect cryptosporidium oocytes, modified acid-fast technique we used.&#13; Results: 12 (11/5%) out of the 104 cases were proved infected by cryptosporidium. This figure dropped to 3.9% in control group respectively. Based on X2 test, rate of infection for the case group was considerably greater than for the control group Infection in the case group didn't show any significant relation with such factors as sex, age and duration of dialysis, history if kidney transplantation and history of immunosuppressive drugs consumption. The rate of infection peaked in diabetic cases (19.4%), while compared with non- diabetic ones (8.3%) (p&lt;0.05).&#13; Conclusions: Our study showed that the prevalence rare of cryptosporidium infection in dialyzed patients was considerably higer than general population. The results were fully correspondent with those of other surveys on immunocompromized patients. Neither Our study nor the similar previously conducted ones revealed no significant association between age and sex and cryptosporidium infection.&#13; We couldn't show any relation between the rate of infection and duration of dialysis.&#13; This difference is likely due to either limited number of cases or a lack of proportional increase in susceptibility to infection atter a certain period of dialysis. Finally we found out that the rate of infection in dialyzed diabetic patients was extra ordinarily higher, which could be the product of acquired immunodeficiency of diabetes

Genotypes Identification of echinococcus granulosus isolated from iranian dogs and camels using three polymerase Chain reaction-based methods of cox1 gene

Aims: Hydatidosis is an important zoonotic disease that is caused by a tiny tapeworm, namely Echinococcus granulosus. In this study, three polymerase chain reaction (PCR)-based methods, including, high resolution melting (HRM) analysis, DNA sequencing, and PCR-restriction fragment length polymorphism (RFLP) have been used for genotype the identification of E. granulosus isolates from dogs and camels in Zarinshahr and Najafabad, Isfahan province, Iran. Materials and Methods: A total of 200 adult worms of 40 dogs and 51 samples of camel hydatid cysts were examined. Molecular characterization of isolates was performed using HRM assay, sequencing of DNA, and digestion Rsa1 pattern coding for the mitochondrial cox1 gene. For analysis of the HRM melting curve, we used the Tm within the range of 77.50°C–79.23°C. Results: HRM analysis revealed that 72.5%, 15%, and 12.5% dog's genotypes and 41.17%, 21.56%, and 35.29% camel genotypes were G1, G3, and G6, respectively. PCR-RFLP analysis, spare parts 310 bp and 138 bp of cox1 that shows the G1 genotype in all of the isolates. Sequence analysis as well as HRM assay was confirmed genotypes of G1, G3, and G6 in camels and dogs. Based on three methods of the cox1 gene the dominant genotype was G1. Conclusion: The PCR-RFLP only identified the G1 genotype, whereas the HRM analysis, as well as DNA sequencing, were detected three genotypes G1, G3, G6, therefore, these two methods have enough accuracy for the determination of genotypes of E. granulosus. This information leads to a better understanding of the biological characteristics of E. granulosus genotypes in Iran and shows the camel as a source of human hydatidosis

EFFECT OF ASCORBIC ACID ON THE RIGHT AND LEFT CORONARY ARTERIES OF MALE RABBITS FED WITH HIGH-CHOLESTEROL DIET

&nbsp; Abstract INTRODUCTION: Atherosclerosis and cardiovascular diseases are the most common causes of death in western countries. The beneficial effect of ascorbic acid on various organs has been reported. The present study was conducted to determine the effect of ascorbic acid on the right and left coronary arteries of male rabbits fed with high-cholesterol diet. methods: Twenty white male rabbits (mean weight: 950 g) were weighed and randomly divided into two groups. For 40 days, group 1 (n=10) was given a high-cholesterol (1%) diet, group 2 (n=10) was fed with a high-cholesterol diet and ascorbic acid (100 mg/kg). Then both of the groups were weighed and the animals were sacrificed. The right and left coronary arteries were dissected and then fixation, tissue processing, histological sectioning and H &amp; E staining were carried out and sections were studied by light microscopy. The results were analyzed by using the Mann Whitney test. results: Group 2 which received ascorbic acid had no fatty streaks in their coronary arteries. Significant difference in mean weight was observed before and after the diet in both groups (P&lt;0.05). CONCLUSIONS: Histopathological study of the coronary arteries showed that the rabbits which received ascorbic acid diet did not develop fatty streaks. Thus ascorbic acid exerts an apparently inhibitory effect on fatty streak formation and may slow down or prevent atherosclerosis by countering the side effects of a high-fat meal. &nbsp; &nbsp; Keywords: Ascorbic acid, cholesterol, coronary arteries.</div

Molecular and genotyping identification of Echinococcus granulosus from camel and dog isolates in Isfahan, Iran (2015-2016)

Background: Hydatidosis is one of the most important zoonotic parasitic disease worldwide caused by Echinococcus granulosus. Given the importance of parasite genotyping in disease control, this study was aimed to evaluate the genetic variation of the camel and dog isolates in Isfahan. Material and Methods: This cross-sectional study was done on infected livers and lungs of camels with hydatid cyst (n=51) during 2015-2016, collected from Zarinshahr and Najafabad industrial abattoir and 200 adult E. granulosus worms collected from stray dogs in different geographical regions of Isfahan for the molecular identification of mitochondrial genes (CO1, ND1). The DNA of protoscolices and adult worms were extracted, the PCR and sequencing were done using specific primers. Pairwise sequence alignment was used to identify the genetic similarity between the sequences and the registered ones in Gen bank using Clustalw2 software. Results: The PCR and sequencing of CO1 and ND1 genes on dog and camel samples showed different E. granulosus genotypes, including the common sheep strain (G1), bovid strain(G3) and camel strain (G6) in Isfahan region. The most frequent strains were the common sheep strain (G1). The genetic similarity between both ND1 and CO1 isolates taken from camel, dog and the isolates registered in Gene bank were 30.25-100% and 27.47-100% for camel and dog, respectively. Conclusion: Mitochondrial ND1 and CO1 genes are proper molecular markers for identifying the genetic variation of the camel and dog E. granulosus. The dominant strain of the worm in Isfahan, is the same as the other parts of the country, i.e., the G1 genotype (the most common sheep strain) belonged to the G1- G1 E. granulosus complex

Synthesis and In vitro Leishmanicidal Activities of Six Quercetin Derivatives

Background: Today, leishmaniasis is a widespread, infectious parasitic disease caused by Leishmania spp. Natural-derived compounds are likely to provide a valuable source of new pharmaceuticals, and among them, quercetin derivatives may have antileishmanial effects. The antileishmanial activity of 3,5,7,3',4'-pentahydroxyflavonol (quercetin) derivatives is partly attributed to the position and pKa of phenolic or catechol hydroxyl groups. Therefore, to optimize their leishmanicidal effect, the structural features of quercetin and its derivatives were improved by acylation or alkylation of hydroxyl groups and changing their pKa and consequently their activities. Materials and Methods: In this study, during a regioselective method, quercetin derivatives were synthesized. The structures of synthesized compounds were confirmed by mass, IR, 1H-, and 13C-NMR spectral data. The antileishmanial activities of compounds 1–6 were compared with glucantime as the standard drug against promastigotes of Leishmania major using standard cell-based leishmanicidal assay. Results: In this study, during a regioselective method, two 7-O-quercetin derivatives (5 and 6), and three quercetin acetate derivatives (2, 3, and 4) were synthesized. In detail, the IC50values found against L. major were (1) 2.5 ± 0.92; (2) 2.85 ± 0.99; (3) 15.5 ± 1.95; (4) 13.5 ± 3.5; (5) 2.6 ± 0.57; and (6) 1.3 ± 0.35 μM while IC50value of glucantime as the standard drug was 88.5 ± 9.47 μM. Conclusions: The present study showed an effective antileishmanial activity of quercetin semisynthetic compounds (1–6) against in vitro promastigotes of L. major. Among them, quercetin analogs with more lipophilic and iron-chelating activity showed more antiparasite activity