Immunoreactivity analysis of Toxoplasma gondii recombinant antigen rSAG3 in sera from immunized BALB/c mice and tox-oplasmosis patients

Background: The coccidian protozoa Toxoplasma gondii is an obligate intracellular parasite of humans and other warm-blooded animals. Diagnosis of toxoplasmosis is of considerable medical importance for human, especially pregnant women and immunocompromised individuals. The apply of an Escherichia coli recombinant antigen(s) would be signifi-cantly useful in developing standardization of the diagnostic tests and reducing their costs. In this study, immunoreac-tivity of recombinant SAG3 against sera from immunized mice and human anti-T. gondii IgG positive patients was evaluated by western-blotting and enzyme immunoassay (EIA) in Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences in 2013. Methods: Three inbreed BALB/c female mice were obtained. Two mice were injected with rSAG3 and one was re-mained untreated, as control. Sera from immunized mice and also pooled sera from IgG positive toxoplasmosis cases were evaluated with western-blotting. IgG antibody responses to recombinant SAG3 was measured by indirect ELISA against the negative control group. Results: The rSAG3 protein reacted with sera of immunized mice and sera from patients with anti-Toxoplasma IgG antibodies in western-blot analysis. The result of ELISA showed that, there was marked differences in the absorbance values between the recombinant SAG3 immunized mice and control group. Conclusion: The rSAG3 showed IgG reactivity with sera from immunized mice and anti-Toxoplasma IgG patients. © 2016, Iranian Journal of Public Health. All rights reserved

A study on prevalence of some helminthic infections of the liver and lungs among ruminants in abattoir of Fars province, Iran

Zoonotic helminths are often transmitted to humans through domestic animals. This retrospective study was performed to estimate the prevalence of some important zoonotic helminthic infections of the liver and lung including echinococcosis, fasciolosis and dicrocoeliasis in Kazerun and Shiraz abattoirs, Fars, Iran, during 2011-2013. A total of 12381 sheep, 6473 cattle, 22847 goats, 66 camels, and 10 buffalos were analyzed with regard to liver and lung helminthic infections including hydatidosis, fasciolosis, and dicrocoeliasis in Kazerun, during March 2011-January 2013. Moreover, 121100 sheep, 23515 cattle, 81293 goats, and 69 camels were investigated for the mentioned infections in Shiraz abattoir during one year since March 2012. Kazerun abattoir: The prevalence rates of liver hydatid cysts were 0.86, 2.1, 0.76, and 15.1 in sheep, cattle, goats, and camels, respectively, whereas the prevalence rates of pulmonary hydatid cysts were 0.89, 2.36, 0.9, and 16.6 in the mentioned animals, respectively. Fasciolosis was detected in 0.33, 1.65, and 0.24 of condemnation livers of sheep, cattle, and goats, respectively. However, dicrocoeliasis was just prevalent in 0.004 of goat livers. Shiraz abattoir: the prevalence rates of liver hydatid cysts were 3.44, 3.12, 2.94, and 2.9 in slaughtered sheep, cattle, goats, and camels, respectively. In addition, 4.54, 4.33, 4, and 4.35 of the lung of the mentioned animals were infected with hydatid cysts, respectively. Prevalence rates of Fasciola spp in slaughtered sheep and cattle were 2.49 and 1.86, respectively, and rate of D. dendriticum infection in slaughtered sheep, cattle, and goats were 0.026, 0.91, and 4, respectively. As compared with reports from other studies, it seems that Fars is among the low-endemic regions regarding this type of infection. © 2016 by Razi Vaccine and Serum Research Institute

Comparison of clinical findings and rapid streptococcal antigen detection test in the diagnosis of group a Streptococcal (GAS) pharyngitis

Background: Group A β-hemolytic Streptococcal (GAS) pharyngitis is a common illness in children. Diagnosis and proper treatment of group A streptococcal sore throat is important particularly to prevent non-superlative sequel. Clinical findings continue to be used in differentiating streptococcal infection from viral sore throat. Objectives: The aim of this study was to evaluate the accuracy of clinical findings and rapid test in comparison with culture in the diagnosis of group A Streptococcal (GAS) pharyngitis. Patients and Methods: Ninety-four children between 3 to 16 years, who were referred to the pediatric clinic of Rasoul-e-Akram hospital with clinical findings of fever or sore throat were evaluated from October 2006 to May 2007. Clinical findings were recorded and swabs were taken for group A streptococcal cultures and streptococcal rapid antigen detection test. Analysis of statistical significance was performed using the chi-square method. The accuracy of clinical findings and rapid test was compared with the culture method as the gold standard, and sensitivity, specificity, positive predictive values, negative predictive value, positive likelihood ratio (LR+) and negative likelihood ratio (LR-) were calculated. Results: The culture was positive in 38 (40.4) of the 94 evaluated children. The mean age of children was 8 ± 3.7 years. The presence of petechiae, exudate and Lymphadenopathy (LAP) was more likely in children with positive streptococcus culture and rapid test (P value < 0.05). Lymphadenopathy was known to feature the most sensitivity (100), specificity (76.8), and positive predictive value (74.5), negative predictive value (100) and positive likelihood ratio (LR) (4.3) among clinical findings. The results of rapid test showed sensitivity of 89.4, specificity of 100, positive predictive value of 100, and negative predictive value of 93.3 in comparison with culture as the gold standard. In general, the accuracy of rapid test was found higher than subjective clinical findings (P = 0.001) Conclusions: Although LAP had good performance in early diagnosis of GAS, a combination of clinical findings, including tonsillar exudates, petechiae with results of rapid antigen test or culture is necessary for clinician judgment. Throat culture is the gold standard test for detecting group A Streptococcal infection, but rapid test is a good replacement for culture. © 2016, Infectious Diseases and Tropical Medicine Research Center

Prevalence of intestinal parasitic infection in cancer, organ transplant and primary immunodeficiency patients in Tehran, Iran

Background: Intestinal parasitic infection in immunodeficient patients especially those with impaired cellular immunity, like neoplasia, renal or heart transplant needs careful consideration. The objective of this study is to evaluate the prevalence of intestinal parasites in different group of patients including cancer patients; organ transplants recipients, and primary immunodeficiency patients. Methods: Stool samples from 190 patients including 80 patients with Primary Immunodeficiency, 85 cancer patients and 25 organ transplant recipients were collected; a direct examination with Phosphate buffered saline (PBS) and formalin ether concentration was performed. The DNA was extracted from parasitologically confirmed patients and nested PCR and sequencing was performed and new obtained sequences of Cryptosporidium parvum and Enterocytozoon bieneusi were compared with deposited ones. Results: In general, the prevalence of parasites was 26/80 (32.5) in primary immunodeficiency, 22/85(25.9) in cancer group, and 7/25 (28) in organ transplant. The prevalence of intestinal parasitic infections in primary immunodeficiency patients were Blastocystis hominis 13 (16.2), Giardia lamblia 10 (12.5), Cryptosporidium 1(1.2), Chilomastix mesnilii 1 (1.2), Dientamoeba fragilis 1(1.2). Of 25 organ transplants, 6 (24) Cryptosporidium sp were found, all of which were confirmed as Cryptosporidium parvum and one case of Microspora in a heart transplant recipient was confirmed as Enterocytozoon bieneusi by PCR sequencing. The predominant intestinal parasitic infection in cancer patients was 19 (22.3) Blastocystis hominis followed by two (2.3) Giardia lamblia and one Dientamoeba fragilis 1 (1.1). Conclusion: The high rate of infection with Blastocystis hominis was found in cancer patients especially colorectal cancer patients, so careful consideration should be given by physicians. Cryptosporidium sp was found to be the major cause of parasitic intestinal infection in patients with organ transplant compared to primary immunodeficiency patients; so transplant recipients undergoing immunosuppressive therapy should be considered as a risk group for acquiring microsporidiosis and Cryptosporidium infection. © 2019, Asian Pacific Organization for Cancer Prevention

In vitro effects of ketotifen and cromolyn sodium on promastigotes and amastigotes of leishmania major

Background: The first line treatment against cutaneous leishmaniasis is meglumine antimoniate. This drug is expensive and has serious side effects, including development of drug resistance. Objectives: In this research, because of paucity of information, the apoptotic and leishmanicidal effects of ketotifen and cromolyn sodium, as cell membrane stabilizer drugs, were investigated on standard strain of Leishmania major. Methods: In this experimental study, L. major parasites were first cultured in RPM1 1640 media, supplemented with 10 fetal bovine serum (FBS) and antibiotics at 24 ± 1°C. Drug concentrations of 5, 10, 15, and 20 µg/mL were then added to L. major culture at 24-, 48- and 72-hour intervals. The 3 - (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) tetrazolium assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Halfmaximal inhibitory concentration (IC50) were ascertained by counting the parasites. The inhibitory effect of the drugs were compared with that of glucantime. Flow cytometry was performed in the next step, to evaluate apoptosis. Each test was repeated three times. Results: IC50 values of ketotifen and cromolyn sodium after 72 hours were calculated to be 2.04 and 17.67 µg/mL for promastigotes and 0.12 and 14.79 µg/mL for amastigotes, respectively. The results of MTT assays showed 20 and 35 promastigote viability after 72 hours of exposure to ketotifen and cromolyn sodium at 20 µg/mL concentration. Apoptosis in ketotifen and cromolyn sodium was quantified to be 11.52 and 9.96 in promastigotes and 99.5 and 98.6 in amastigote-infected macrophages, respectively. The results indicated that the drugs induce early and late apoptosis in parasites. All treatments produced results, which differed significantly from the control groups (P < 0.05). Conclusions: Drugs used in this study, especially Ketotifen, showed lower toxicity yet similar anti-leishmanial effectsonboth forms, as cromolyn sodium did. It could be suggested that further investigations about the in vivo effects of these drugs, as candidates for cutaneous leishmaniasis treatment, are required. © 2018, Author(s)

Detection of pseudocyst forms of trichomonas muris in rodents from iran

Background: Trichomonas muris is one of the most common protozoa diagnosed in rodents. The trichomonads are generally described as presenting only trophozoite form while pseudocyst is another morphological form of trichomonads identified among gastrointestinal and genitourinary trichomonads. We identified and described different shapes of T. muris pseudocysts and trophozoite in stool samples were collected from rodents including Merinos persicus, Mus musculus and Cricetulus migratorius. Methods: In this cross-sectional study, stool samples from 204 trapped rodents were collected from Meshkin Shahr during Mar to Dec 2014. Samples were preserved in formalin 10 and PVA solution and transferred to Department of Medical Protozoology and Mycology, School of Public Health, Tehran University of Medical Sciences. Formalin-ether concentration method was used for the samples. The slides were stained with trichrome staining method and observed under light microscope. Results: The trophozoites were classified as T. muris based on size (18 to 24 µm), presence of three anterior flagella, recurrent flagellum, undulating membrane, and axostyle in direct examination and stained slides with trichrome staining method. Fifty-five out of 204 (27) rodents were infected with T. muris in which 51(25) samples pseudocysts form were observed. The spherical bodies of pseudocyst with almost 8 µm size, contained internalized flagella, an undulating membrane with recurrent flagellum, axostyle, and costa were seen. The pseudocysts were more prevalent than trophozoite form and pseudocysts were found with different shapes in this study. Conclusion: T. muris pseudocysts were found in stool samples of caught rodents for the first time in northwestern Iran. © 2018, Iranian Journal of Public Health. All rights reserved

Detection of pseudocyst forms of trichomonas muris in rodents from iran

Background: Trichomonas muris is one of the most common protozoa diagnosed in rodents. The trichomonads are generally described as presenting only trophozoite form while pseudocyst is another morphological form of trichomonads identified among gastrointestinal and genitourinary trichomonads. We identified and described different shapes of T. muris pseudocysts and trophozoite in stool samples were collected from rodents including Merinos persicus, Mus musculus and Cricetulus migratorius. Methods: In this cross-sectional study, stool samples from 204 trapped rodents were collected from Meshkin Shahr during Mar to Dec 2014. Samples were preserved in formalin 10 and PVA solution and transferred to Department of Medical Protozoology and Mycology, School of Public Health, Tehran University of Medical Sciences. Formalin-ether concentration method was used for the samples. The slides were stained with trichrome staining method and observed under light microscope. Results: The trophozoites were classified as T. muris based on size (18 to 24 µm), presence of three anterior flagella, recurrent flagellum, undulating membrane, and axostyle in direct examination and stained slides with trichrome staining method. Fifty-five out of 204 (27) rodents were infected with T. muris in which 51(25) samples pseudocysts form were observed. The spherical bodies of pseudocyst with almost 8 µm size, contained internalized flagella, an undulating membrane with recurrent flagellum, axostyle, and costa were seen. The pseudocysts were more prevalent than trophozoite form and pseudocysts were found with different shapes in this study. Conclusion: T. muris pseudocysts were found in stool samples of caught rodents for the first time in northwestern Iran. © 2018, Iranian Journal of Public Health. All rights reserved

Intestinal protozoa in domestic cats (Carnivora: Felidae, felis catus) in northwestern iran: A cross-sectional study with prevalent of microsporidian and coccidian parasites

Background: In this study, some microsporidial and coccidian parasites were isolated from 103 domestic cats in the Meshkin Shahr area, northwestern Iran during the Jun 2014 to Jun 2015, and their genera were identified using parasitologicamethods with emphasis on their zoonotic importance. Methods: One hundred and three fecal samples of domestic cats were collected and preserved in formalin (10) and conserved in phosphate buffer saline solution, finally examined by microscopy after formalin-ether concentration and specific staining. Preservation in dichromate potassium (2.5) was performed for all coccidian positive samples and then sporulated coccidian oocysts were investigated. Results: The detected parasites were Isospora spp. 6/103(5.8). Microsporidian spores were identified in 46/103 (44.6) of all samples post-stained by the aniline blue staining method. Conclusion: Microsporidial infections were more prevalent in domestic cats. Further studies are needed in the identification of microsporidial spores isolated from infected cats. © 2019, Tehran University of Medical Sciences (TUMS). All rights reserved

Acute and post-acute phase of COVID-19: Analyzing expression patterns of miRNA-29a-3p, 146a-3p, 155-5p, and let-7b-3p in PBMC

Background: When a new pathogen, such as severe acute respiratory syndrome coronavirus 2, appears all novel information can aid in the process of monitoring and in the diagnosis of the coronavirus disease (COVID-19). The aim of the current study is to elucidate the specific miRNA profile which can act as new biomarkers for distinguishing acute COVID-19 disease from the healthy group and those in the post-acute phase of the COVID-19 disease. Methods: The expression level of selected miRNAs including let-7b-3p, miR-29a-3p, miR-146a-3p and miR-155-5p were evaluated in peripheral blood mononuclear cells (PBMCs) of COVID-19 patients, in both the acute and post-acute COVID-19 phase of the disease and healthy groups, by real-time PCR assays. Specificity and sensitivity of miRNAs was tested by receiver operating characteristic (ROC) analysis in COVID-19 patients. Results: The expression level of all miRNAs in COVID-19 patients was significantly higher than in the healthy group. Therefore, the expression pattern of miR-29a-3p, miR-146a-3p and let-7b-3p in the post-acute COVID-19 phase was significantly different from the acute COVID-19 phase. ROC analyses demonstrated that miR-29a-3p, -155-5p and -146a-3p may serve as the novel biomarker for COVID-19 diagnosis with high specificity and sensitivity. In addition, miR-29a-3p, and -146a-3p can maybe act as novel biomarkers for distinguishing acute from post-acute phase of COVID-19 disease. Discussion: The difference in miRNA expression pattern between COVID-19 patients and those in the healthy group, and between acute COVID-19 with post-acute COVID-19, suggested that cellular miRNAs could be used as promising biomarkers for diagnosis and monitoring of COVID-19. © 2021 Elsevier B.V

Comparison of serologic status of Toxoplasma gondii infection in pre- and post-heart transplantation in a pediatric population: A preliminary study

Background: Toxoplasmosis is an important opportunistic infection in immunocompromised children, especially in heart transplant recipients. This study aimed to investigate pre- and post-transplant serology for toxoplasmosis along with post-transplant PCR in pediatric heart transplant patients. Methods: This cross-sectional study was performed on 38 heart transplant recipients aged 1-17 years, by the end of 2018. Pre- and post-transplant IgM and IgG titrations were measured using ELISA method. Nested PCR of B1 gene was performed to identify Toxoplasma gondii (T gondii) infection after transplant. Results: Totally, 11.4 of patients had positive IgG and 91.4 had negative IgM for toxoplasmosis before heart transplantation. The mean of pre-transplant IgG titration for seropositive and seronegative patients was 22.32 ± 15.30 IU/mL and 1.49 ± 1.15 IU/mL, respectively (P  12 months (1.07 ± 1.27 IU/mL; P =.004) time periods. The result of PCR for B1 gene in all cases was negative. Conclusions: Chemoprophylaxis with TMP/SMX seems to be effective in prevention of T gondii infection or reactivation among pediatric heart transplantation population. Anti-T. gondii-IgG level alone may not be sensitive enough for evaluation of the infection at least after 6 months post-transplantation. © 2020 Wiley Periodicals LL