Publications by authors named "Gholam Reza Hatam"

36 Publications

HLA, Immune Response, and Susceptibility to COVID-19.

Front Immunol 2020;11:601886. Epub 2021 Jan 8.

Spondylitis Program, Division of Rheumatology, Schroeder Arthritis Institute, University Health Network, Toronto, ON, Canada.

The severe acute respiratory syndrome caused by Coronavirus 2 (SARS-CoV-2) that appeared in December 2019 has precipitated the global pandemic Coronavirus Disease 2019 (COVID-19). However, in many parts of Africa fewer than expected cases of COVID-19, with lower rates of mortality, have been reported. Individual human leukocyte antigen (HLA) alleles can affect both the susceptibility and the severity of viral infections. In the case of COVID-19 such an analysis may contribute to identifying individuals at higher risk of the disease and the epidemiological level to understanding the differences between countries in the epidemic patterns. It is also recognized that first antigen exposure influences the consequence of subsequent exposure. We thus propose a theory incorporating HLA antigens, the "original antigenic sin (OAS)" effect, and presentation of viral peptides which could explain with differential susceptibility or resistance to SARS-CoV-2 infections.
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http://dx.doi.org/10.3389/fimmu.2020.601886DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7820778PMC
February 2021

Cytokine profile and nitric oxide levels in peritoneal macrophages of BALB/c mice exposed to the fucose-mannose ligand of Leishmania infantum combined with glycyrrhizin.

Parasit Vectors 2020 Jul 20;13(1):363. Epub 2020 Jul 20.

Faculty of Medicine and Health Technology, Tampere University, 33014, Tampere, Finland.

Background: The fucose-mannose ligand (FML) of Leishmania infantum is a complex glycoprotein which does not elicit adequate immunogenicity in humans. In recent years, adjuvant compounds derived from plants have been used for improving the immunogenicity of vaccines. Glycyrrhizin (GL) is a natural triterpenoid saponin that has known immunomodulatory activities. In the present study, we investigated the effects of co-treatment with FML and GL on the production of cytokines and nitric oxide (NO) by macrophages, in vitro.

Methods: Lipopolysaccharide (LPS) stimulated murine peritoneal macrophages were treated with FML (5 μg/ml) of L. infantum and various concentrations of GL (1 μg/ml, 10 μg/ml and 20 μg/ml). After 48 h of treatment, cell culture supernatants were recovered and the levels of TNF-α, IL-10, IL-12p70 and IP-10 were measured by sandwich ELISA and NO concentration by Griess reaction.

Results: Our results indicate that the treatment of activated macrophages with FML plus GL leads to enhanced production of NO, TNF-α and IL-12p70, and reduction of IL-10 levels in comparison with FML treatment alone.

Conclusions: Therefore, we concluded that GL can improve the immunostimulatory effect of FML on macrophages and leads to their polarization towards an M1-like phenotype.
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http://dx.doi.org/10.1186/s13071-020-04243-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7370265PMC
July 2020

Isoenzyme profiles and phylogenetic analysis of Giardia duodenalis isolates from Iranian patients.

Environ Sci Pollut Res Int 2020 Nov 15;27(32):40652-40663. Epub 2020 Jul 15.

Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

The main objective of this study was to characterize the Giardia duodenalis isolates from Iranian patients in Fars Province, south of Iran by biochemical and molecular methods. Fifteen mass cultivated of G. duodenalis isolates in modified TYI-S-33 medium were analyzed using isoenzyme electrophoresis and PCR genotyping. Polyacrylamide gel electrophoresis (PAGE) of five different enzyme systems was used to characterize isolates: (i) glucose-6-phosphate dehydrogenase, (ii) glucose phosphate isomerase, (iii) malate dehydrogenase, (iv) malic enzyme, and (v) phosphoglucomutase. As well, a fragment of the SSU-rDNA (292 bp) gene was amplified by PCR using the primers RH11 and RH4. The sequencing of the PCR products and phylogenetic tree were performed. The isoenzyme electrophoretic profiles divided fifteen G. duodenalis isolates into four zymodemes. G6PD, GPI, MDH, ME, and PGM enzyme systems showed 1, 2, 2, 3, and 3 enzyme pattern, respectively. G6PD isoenzyme pattern had the most homogeneity, while isoenzyme patterns of ME and PGM had the most heterogeneity in our study. Genotyping results indicated that the zymodemes 1-4 were categorized in assemblage A based on the SSU-rDNA gene. Phylogenetic analysis showed that all four zymodemes were distributed within the cluster of assemblage A. Our results indicated that both isoenzyme and DNA analyses were useful to characterize the isolates of Giardia and distinguishing various zymodemes and assemblages. It could be suggested that the genetic diversity among isoenzymes profiles of G. duodenalis may explain the variable clinical manifestations, pathogenicity, host response, drug susceptibility, and treatment efficacy of human giardiasis.
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http://dx.doi.org/10.1007/s11356-020-10062-1DOI Listing
November 2020

A new multi-epitope peptide vaccine induces immune responses and protection against Leishmania infantum in BALB/c mice.

Med Microbiol Immunol 2020 Feb 6;209(1):69-79. Epub 2019 Nov 6.

Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Visceral leishmaniasis (VL) is a tropical and subtropical disease which is endemic in more than eighty countries around the world. Leishmania infantum is one of the main causative agents of VL disease. Currently, there is no approved-to-market vaccine for VL therapy. In this study, we evaluated cellular and humoral immune responses induced by our newly designed multi-epitope vaccine in BALB/c mice. Four antigenic proteins, including histone H1, sterol 24-c-methyltransferase (SMT), Leishmania-specific hypothetical protein (LiHy), and Leishmania-specific antigenic protein (LSAP) were chosen for the prediction of potential immunodominant epitopes. Moreover, to enhance vaccine immunogenicity, two toll-like receptors 4 (TLR4) agonists, resuscitation-promoting factors of Mycobacterium tuberculosis (RpfE and RpfB), were employed as the built-in adjuvants. Immunization with the designed multi-epitope vaccine elicited a robust Th1-type immune response, compared to other groups, as shown by increased levels of IL-2, IFN-γ, TNF-α, and IgG2a. Furthermore, a significant decrease was observed in Th-2-type-related cytokines such as IL-4 in immunized mice. The designed construct also induced a significant reduction in parasite load (p < 0.0001), conferring protection against L. infantum challenge. This study could be promising in gaining insight towards the potential of peptide epitope-based vaccines as effective protective approaches against Leishmania species.
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http://dx.doi.org/10.1007/s00430-019-00640-7DOI Listing
February 2020

Investigation of the Physical, Chemical Characteristics and Microbial Contamination of the Indoor Swimming Pools

Turkiye Parazitol Derg 2019 Sep;43(3):130-134

Shiraz University of Medical Sciences, Prof. Alborzi Clinical Microbiology Research Center, Shiraz, Iran

Objective: The aim of this study was to investigate the physical, chemical and microbiological contamination of indoor swimming pools.

Methods: Pool water specimens were collected using a plastic polypropylene sterilized bottle. The physical and chemical qualities of the waters were analyzed in terms of temperature, turbidity, pH, and free residual chlorine, with the standard methods for the examination of water. Bacteriological (routine methods) and parasitological (molecular methods) tests were carried out on pools water.

Results: The mean temperature, pH, and residual chlorine of the indoor pools were 31.2 °C, 7.6 and 1.5 mg/L, respectively. Turbidity was not observed in any of the pools. The pH and temperature values were in standard ranges in 92.3% and 15.4% of the waters of swimming pools, respectively. The prevalence rates of bacterial and amoebic contaminations of the water in the swimming pools were 53.8% and 46.2%, respectively. One pool (7.7%) was contaminated with both bacteria and amoeba. and spp. were isolated from the pool waters.

Conclusion: In this study, some microorganisms were identified from the water pools. Effective management of swimming pools and proper control of the physical, chemical and microbiological property of water pools can produce the healthy recreational activity.
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http://dx.doi.org/10.4274/tpd.galenos.2019.6112DOI Listing
September 2019

Leishmania cytochrome b gene sequence polymorphisms in southern Iran: relationships with different cutaneous clinical manifestations.

BMC Infect Dis 2019 Jan 29;19(1):98. Epub 2019 Jan 29.

Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Background: Cutaneous leishmaniasis (CL) caused by Leishmania species, is a geographically extensive disease that infects humans and animals. CL is endemic in half of the 31 provinces of Iran, with 29,201 incidence cases reported in Fars province from 2010 to 2015. CL is polymorphic and may result in lesions characterized by different clinical features. Parasite genetic diversity is proposed to be one of the factors affecting the clinical outcome and lesion characteristics in CL patients. However, there is still very limited data regarding the genetic variation of Leishmania spp. based on the sequencing of Cytochrome b (Cyt b) gene.

Methods: All patients originated from endemic regions in Fars province. The amplification of the Cyt b gene from isolates of 100 patients with disparate clinical forms of CL was accomplished using Nested-PCR. Sequence analysis of the amplified Cyt b was used to scrutinize the genetic variations among Leishmania isolates and connect the results with clinical pictures. The clinical demonstrations were basically of two types, typical and atypical lesions. Molecular phylogenetic tree was constructed using the Neighbor-Joining method, with species/strains from this study compared to species/strains from other geographical regions.

Results: Leishmania major was identified as the predominant infecting Leishmania spp. (86% of cases), with the remainder of cases being infected by Leishmania tropica. Clinical examination of patients revealed 12 different clinical CL forms. Among Leishmania samples analyzed, five distinct haplotypes were recognized: three in L. major and two in L. tropica. We found a correlation between clinical outcomes and Cyt b sequence variation of Leishmania spp. involved. Moreover, we observed a higher presence of polymorphisms in L. major compared with L. tropica. This difference may be due to the different eco-epidemiologies of both species, with L. tropica being an anthroponosis compared to L. major, which is a zoonosis.

Conclusions: The sequence analysis of Cyt b gene from 25 L. major and L. tropica strains demonstrated genetic variability of L. major and L. tropica causing CL in southern Iran, and a feasible connection amid the genetic heterogeneity of the parasite, geographical source and clinical appearance of the disease in human was detected.
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http://dx.doi.org/10.1186/s12879-018-3667-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6352432PMC
January 2019

Immunoinformatics-aided design of a potential multi-epitope peptide vaccine against Leishmania infantum.

Int J Biol Macromol 2018 Dec 30;120(Pt A):1127-1139. Epub 2018 Aug 30.

Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran. Electronic address:

Visceral leishmaniasis (VL) or kala-azar, the most severe form of the disease, is endemic in more than eighty countries across the world. To date, there is no approved vaccine against VL in the market. Recent advances in reverse vaccinology could be promising approach in designing the efficient vaccine for VL treatment. In this study, an efficient multi-epitope vaccine against Leishmania infantum, the causative agent of VL, was designed using various computational vaccinology methods. Potential immunodominant epitopes were selected from four antigenic proteins, including histone H1, sterol 24-c-methyltransferase (SMT), Leishmania-specific hypothetical protein (LiHy), and Leishmania-specific antigenic protein (LSAP). To enhance vaccine immunogenicity, two resuscitation-promoting factor of Mycobacterium tuberculosis, RpfE and RpfB, were employed as adjuvants. All the aforesaid segments were joined using proper linkers. Homology modeling, followed by refinement and validation was performed to obtain a high-quality 3D structure of designed vaccine. Docking analyses and molecular dynamics (MD) studies indicated vaccine/TLR4 complex was in the stable form during simulation time. In sum, we expect our designed vaccine is able to induce humoral and cellular immune responses against L. infantum, and may be promising medication for VL, after in vitro and in vivo immunological assays.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.08.125DOI Listing
December 2018

A survey of the frequency of cytomegalovirus-associated diarrhea in immunocompromised patients using a non-invasive method.

Iran J Microbiol 2018 Apr;10(2):143-150

Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Background And Objectives: infection is the most common viral opportunistic infection causing gastrointestinal diseases such diarrhea and colitis in immunocompromised patients. The development and performance of a robust and sensitive PCR assay are usually evaluated to detect CMV DNA in human fecal specimens. In this study, our aim was to detect CMV DNA in stool samples taken from patients with HIV/AIDS, cancer, and transplant recipient patients with chronic and persistent diarrhea using a non-invasive method.

Materials And Methods: A total of 633 immunocompromised patients (451 males and 182 females) suffering from persistent or chronic diarrhea were included in this study. Among them, 392 were HIV/AIDS patients, 151 had cancer and were receiving chemotherapy, and 90 were recipients of a solid organ or bone marrow transplant. CMV genome was extracted from the stool samples using phenol: chloroform: isoamyl alcohol method. CMV DNA was identified by polymerase chain reaction using sequence specific primers on genomic DNA.

Results: Looking at the frequency of CMV DNA in 392 HIV/AIDS patients, we found that only 5 patients (1.27%) were positive for CMV genome, while this frequency was 4.63% (7/151) and 5.5% (5/90) in patients with cancer receiving chemotherapy and in those with solid organ or bone marrow transplant, respectively.

Conclusion: The results of this study revealed that the cause of chronic or persistent diarrhea in HIV/AIDS, cancer, and graft recipient patients might be related to CMV infection. Accordingly, we recommend a non-invasive method, such as stool sample, as a first line of diagnosis of enteritis when the physician suspects that a patient has CMV infection.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6039454PMC
April 2018

Vaccination with Live Attenuated L. Major and TLR4 Agonist Promotes a Th1 Immune Response and Induces Protection against L. Major Infection in BALB/c Mice.

Iran J Immunol 2018 Jun;15(2):74-83

Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Background: Toll like receptors play a major role in immune responses against Leishmania parasites.

Objective: To evaluate the efficacy of vaccination with live attenuated L. major and TLR4 agonist in protection against L. major infection.

Methods: Attenuated L. major was prepared by continuous sub-culturing of the parasite. A total of 90 mice were assigned to 9 groups including 6 groups of BALB/c (G1-6) and 3 groups (G7-9) of C57BL/6 mice. Group 1 was the control groups, group 2 received the wild-type L. major promastigotes, group 3 the attenuated line, group 4 the TLR4 agonist, group 5 the wild-type L. major and TLR4 agonist, and group 6 the attenuated line along with TLR4 agonist. Group 7 was control, group 8 received wild-type L. major and group 9 the wild-type along with TLR4 agonist. Vaccinated mice were then challenged with wild-type of L. major. Lesion size, parasite burden, and the expression levels of IL-4, IFN-γ, IL-2, 1L-17A, IL-10, TGF-β and TLR4 were evaluated before the challenge while parasite burden and lesion size were evaluated.

Results: Vaccinated mice with a TLR4 agonist or attenuated L. major plus TLR4 agonist produced the highest levels of IFN-γ, IL-2, and IL-17A. Post-challenge analysis revealed that mice vaccinated with the attenuated line along with TLR4 agonist displayed the lowest lesion size and parasite load. These mice developed a predominant Th1 immune response.

Conclusion: Vaccination with the attenuated L. major along with TLR4 agonist promotes a Th1-mediated immune response which leads to the protection of BALB/c mice against L. major infection.
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http://dx.doi.org/IJIv15i2A1DOI Listing
June 2018

Asymptomatic Infected Children: A Seroprevalence and Molecular Survey in a Rural Area of Fars Province, Southern Iran.

J Trop Med 2018 15;2018:8167247. Epub 2018 May 15.

Department of Parasitology and Mycology, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran.

The current study aimed to evaluate the seroprevalence of visceral leishmaniasis in asymptomatic healthy children in a rural area of Fars province, Southern Iran. Blood samples were taken from 617 asymptomatic healthy children and serum samples along with buffy coat were separated from the blood. The serum samples were assessed for antibodies against by an indirect ELISA and the buffy coats were tested for the presence of DNA by molecular method. Of the 617 recruited children, 297 (48.1%) were female and 317 (51.4%) were male. Anti- antibodies were detected in 17 (2.8%) of the children. From those 17 seropositive cases, 5 (29.4%) were male and 12 (70.6%) cases were female. Children aged 5-8 years had the highest seroprevalence rate; however, no associations were found between seropositivity to and gender or age of the children. Moreover, DNA was detected in buffy coat of 8 (1.3%) of 617 children. Three of the PCR-positive cases were seropositive whereas 14 of seropositive subjects (82.3%) were PCR-negative. Findings of the current study revealed a considerable subclinical leishmanial infection in children in the studied rural area in the south of Iran. Results of the current study could be used for surveillance, prevention, and control of VL in the area.
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http://dx.doi.org/10.1155/2018/8167247DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5976912PMC
May 2018

Proteome-scale identification of Leishmania infantum for novel vaccine candidates: A hierarchical subtractive approach.

Comput Biol Chem 2018 Feb 24;72:16-25. Epub 2017 Dec 24.

Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran; Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran. Electronic address:

Vaccines are one of the most significant achievements in medical science. However, vaccine design is still challenging at all stages. The selection of antigenic peptides as vaccine candidates is the first and most important step for vaccine design. Experimental selection of antigenic peptides for the design of vaccines is a time-consuming, labor-intensive and expensive procedure. More recently, in the light of computer-aided biotechnology and reverse vaccinology, the precise selection of antigenic peptides and rational vaccine design against many pathogens have developed. In this study, the whole proteome of Leishmania infantum was analyzed using a pipeline of algorithms. From the set of 8045 proteins of L. infantum, sixteen novel antigenic proteins were derived using a hierarchical proteome subtractive analysis. These novel vaccine targets can be utilized as top candidates for designing the new prophylactic or therapeutic vaccines against visceral leishmaniasis. Significantly, all the sixteen novel vaccine candidates are non-allergen antigenic proteins that have not been used for the design of vaccines against visceral leishmaniasis until now.
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http://dx.doi.org/10.1016/j.compbiolchem.2017.12.008DOI Listing
February 2018

Considerable Genetic Diversity of Clinical Isolates in a Targeted Population in South of Iran.

Iran J Parasitol 2017 Apr-Jun;12(2):251-259

Research Center for Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran.

Background: The present study aimed to characterize genetically and to compare the most frequently occurring strains of isolated from southern Iran.

Methods: Totally, 150 vaginal swab and urine specimens were collected from symptomatic and asymptomatic women from May 2012 to Jun 2013. This study implemented a sensitive and reliable PCR-restriction fragment length polymorphism (RFLP) typing method on the actin gene. Moreover, one representative sample of each identified genotype was subjected to sequencing.

Results: Twenty-four isolates were positive and 6 distinct electrophoretic patterns (H, E, G, I, M, N) were identified. Genotypes H and I were found to be more prevalent (50 and 37.5%) in Kerman and Shiraz, respectively. The phylogenetic analysis showed that two isolates were located as a separated clade with the other isolates.

Conclusion: The obtained findings showed a considerable genetic polymorphism of clinical isolates from the population studied. More studies may be warranted in future as to unveiling any possible links between a given genotype/cluster and pathogenic behavior of
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5527036PMC
August 2017

Cystoisosporiasis-related human acalculous cholecystitis: the need for increased awareness.

Pol J Pathol 2016;67(3):270-276

Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. and

Cholecystitis is one of the common surgical indications affecting human beings in many countries. A variety of infectious agents can be associated with acute or chronic acalculous cholecystitis, especially in HIV/AIDS patients. In this investigation, the authors aim to describe two cases of histologically and molecularly documented cystoisosporiasis (syn. isosporiasis) as the cause of chronic acalculous cholecystitis in two immunodeficient patients. During microscopic examinations of more than 2500 diarrheic patients' samples, 11 cases of cystoisosporiasis-related recurrent persistent/chronic diarrhea were detected. A review of the medical records of Cystoisospora belli (syn. Isospora belli)-positive patients showed that two of them, i.e. a patient with prolonged corticosteroid therapy and an AIDS patient, several months prior to fecal examinations had undergone cholecystectomy due to acalculous cholecystitis. The study was continued by a review of the histopathological investigation of the recuts prepared from the excised gallbladder tissue sections and stained with hematoxylin and eosin in order to detect a possible specific clinical correlation with cystoisosporiasis. Light microscopic examination revealed the presence of various developmental stages of a coccidial parasite, namely Cystoisospora belli, in both patients' gallbladder tissue sections. To the best of our knowledge, C. bellii-associated cholecystitis has not been previously reported in a patient with prolonged corticosteroid therapy.
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http://dx.doi.org/10.5114/pjp.2016.63779DOI Listing
June 2017

Lip leishmaniasis: a case series with molecular identification and literature review.

BMC Infect Dis 2017 01 25;17(1):96. Epub 2017 Jan 25.

Department of Medical Parasitology & Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Background: Mucocutaneous leishmaniasis (MCL), a protozoan infectious disease, is very rare in Iran despite the endemicity of both cutaneous and visceral forms. It is transmitted by the Phlebotomus sand fly. The lip is considered one of the extraordinary sites. Lesions usually initiate with erythematous papules, slowly enlarges and then it ulcerates. The diagnosis of MCL encompasses epidemiological, clinical and laboratory aspects. Usually, the combination of some of these elements is necessary for the final diagnosis. So, lip leishmaniasis lesions can be challenging to diagnose.

Case Presentation: We presented seven rare cases of lip leishmaniasis. Tissue impression smear, culture, PCR and phylogenetic analysis were carried out for explicit diagnosis. Skin scraping investigation showed several Leishmania spp. amastigotes in the cytoplasm of macrophages. Culture examination was positive for Leishmania spp. PCR was positive for L. major, L. tropica, and L. infantum. Differential diagnosis includes orofacial granulomatosis, basal cell carcinoma, squamous cell carcinoma, and mesenchymal tumors. The cases were treated with systemic meglumine antimoniate (Glucantime). No relapses were observed during 1 year of follow-up. Early detection of the infection are necessary in order to start effective treatment and prevent more serious complications.

Conclusions: In this paper, we reported seven rare cases of lip leishmaniasis in Iran, emphasized the importance of clinical and diagnostic features of lesions, characterized the phylogenetic kinship of isolated parasites, and reviewed the literature on lip leishmaniasis.
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http://dx.doi.org/10.1186/s12879-016-2178-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5264488PMC
January 2017

Cutaneous Leishmaniasis of the Eyelids: A Case Series with Molecular Identification and Literature Review.

Korean J Parasitol 2016 Dec 31;54(6):787-792. Epub 2016 Dec 31.

Department of Medical Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Cutaneous leishmaniasis (CL) is a protozoan disease which is endemic in Iran. It is transmitted by the sand fly. The eyelid is rarely involved possibly because the movement of the lids impedes the sand fly from biting the skin in this region. Here, we report 6 rare cases of eyelid CL. The patients were diagnosed by skin scraping, culture, and PCR from the lesions. Skin scraping examination showed spp. amastigotes in the cytoplasm of macrophages. Culture examination was positive for spp. PCR was positive for and . The lesions were disguised as basal cell carcinoma, chalazion, hordeolum, and impetigo. The patients were treated with intramuscular meglumine antimoniate (20 mg/kg/day) for at least 3 weeks. They showed a dramatic response, and the lesions almost completely disappeared. We emphasized the importance of clinical and diagnostic features of lesions, characterized the phylogenetic relationship of isolated parasites, and reviewed the literature on ocular leishmaniasis.
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http://dx.doi.org/10.3347/kjp.2016.54.6.787DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5266367PMC
December 2016

The Sleep in Caenorhabditis elegans: What We Know Until Now.

Mol Neurobiol 2018 01 11;55(1):879-889. Epub 2017 Jan 11.

Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Sleep, as one of the most important requirements of our brain, has a mystical nature. Despite long-standing studies, the molecular mechanisms and physiological properties of sleep have not been defined well as the complexity of the mammals' brain make it difficult to investigate the mechanisms and properties of sleep. Although some features of sleep have changed during evolution, its existence in such a simple animal, Caenorhabditis elegans, not only signifies the importance of sleep in even simple animals, but also allows the scientist to assess the core mechanism and biological events in an uncomplicated organism. This article reviews the information which exists about the characteristics of sleep in C. elegans, its circadian rhythm, the neurons and neurotransmitters responsible for each state, and the signaling molecules involved. Although much still remains to be resolved about the sleep of C. elegans, the available knowledge helps the scientists to recognize the properties better of this mysterious function of the brain.
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http://dx.doi.org/10.1007/s12035-016-0362-9DOI Listing
January 2018

Human intestinal sarcocystosis in Iran: there but not seen.

Parasitol Res 2016 Dec 9;115(12):4527-4533. Epub 2016 Sep 9.

Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Human sarcocystosis is a rare infection caused by the genus Sarcocystis who human serve as definitive (intestinal form of infection) host or intermediate (extraintestinal form) host for some various Sarcocystis species. The detection of Sarcocystis oocysts/sporocysts in the feces usually incidentally and is achieved by microscopic examination of the fresh or preserved specimens. To rule out any parasitological etiology among 23,875 (aged 2 months to 95 years) apparently immunocompetent Iranian individuals (from October of 2010 to June of 2016) with abdominal discomforts referred to several teaching hospitals and local clinical laboratories in Fars Province, Iran, their fecal samples were examined using light microscopy. Most pathogenic parasite-positive and doubtful samples were sent to the Intestinal Protozoology Laboratories of Fasa and Shiraz Universities of Medical Sciences to further examination to detect probable co-infection with other underdiagnose parasitoses. In addition to the common protozoal and helminthic infections, during the course of examining stool specimens using direct smear mixed with saline or iodine mounts and by formalin-ethyl acetate techniques, four cases of intestinal Sarcocystis infection as only or concurrently infected with other parasites were found. The present paper presents cases of human intestinal Sarcocystis infection in Iran. Since Sarcocystis are small in size and usually rare in stool, they often go unnoticed. It should be noted that stool smears must be examined with great care to avoid misinterpretation of Sarcocystis infections in microscopic examinations. To the best of our knowledge, co-infection of intestinal sarcocystosis and other principal parasitoses in stool investigations has not been reported earlier.
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http://dx.doi.org/10.1007/s00436-016-5244-6DOI Listing
December 2016

In vitro efficacy of ethanolic extract of Artemisia absinthium (Asteraceae) against Leishmania major L. using cell sensitivity and flow cytometry assays.

J Parasit Dis 2016 Sep 20;40(3):735-40. Epub 2014 Sep 20.

Department of Medical Entomology and Vector Control, Research Centre for Health Sciences, School of Health, Shiraz University of Medical Sciences, P.O. Box 71645-111, Shiraz, Iran.

Leishmaniasis is one of the most neglected human diseases with an estimated global burden ranking second in mortality and fourth in morbidity among the tropical infections. Chemotherapy involving the use of drugs like glucantime is the mainstay treatment in endemic areas of Iran. Drug resistance is increasingly prevalent, so search for alternative therapy is gathering pace. Medicinal herbs, like wormwood Artemisia, have chemical compounds effective against a number of pathogens. In this study, the efficacy of ethanol extract from Artemisia absinthium (Asteraceae) against Leishmania major L. was investigated in vitro. The outcome of different effective doses (1-40 mg/ml) of ethanol extracts from this medicinal herb, A. absinthium, on a standard Iranian parasite strain of L. major was examined. The L. major promastigote cell sensitivity and mortality or viability effects due to the addition of herbal extract were measured using the MTT assay and the flow cytometry technique, respectively. There was complete agreement between the two assays. The lethal concentration (LC50) was measured as 101 mg/ml. Some contrasting relationships between the medicinal herb concentrations and the viability of parasites were observed; so that there was an increased multiplication of the parasite at low concentrations of the drug, but an anti-parasitic apoptotic effect was seen at high concentrations of A. absinthium. It was concluded that there might be one or more chemical constituents within the herbal extract of wormwood which at high concentration controlled cell division and affected the relevant activity within the only one giant mitochondrion in this flagellate parasite. At low doses, however, it showed the opposite effect of leading to mitotic cell divisions.
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http://dx.doi.org/10.1007/s12639-014-0569-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4996182PMC
September 2016

Prevalence of Enteric Protozoan Oocysts with Special Reference to Sarcocystis cruzi among Fecal Samples of Diarrheic Immunodeficient Patients in Iran.

Korean J Parasitol 2016 Jun 30;54(3):339-44. Epub 2016 Jun 30.

Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.
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http://dx.doi.org/10.3347/kjp.2016.54.3.339DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4977794PMC
June 2016

Correlation of Genetic Heterogeneity with Cytopathological and Epidemiological Findings of Leishmania major Isolated from Cutaneous Leishmaniasis in Southern Iran.

Acta Cytol 2016 24;60(2):97-106. Epub 2016 May 24.

Department of Pathology, School of Veterinary Medicine, Shahrekord University, Shahrekord, Iran.

Objective: Cutaneous leishmaniasis (CL) has recently grown into a major public health problem in 88 countries of the world, including Iran. It is a polymorphic disease which may show various clinicopathological features. Although the effect of the genetic diversity of the parasite has been demonstrated as one of the factors influencing clinical manifestations in CL, no data exist regarding the genetic variation of Leishmania major and its microscopical features.

Study Design: Fine-needle aspiration, touch smears and the histological sections of 100 patients were examined for Leishmania amastigotes, using Giemsa and hematoxylin and eosin. Diverse types of inflammatory cells in the 40 positive and 5 negative smears were differentiated. Kinetoplast DNA (kDNA) was amplified using nested PCR and subsequently sequenced. Sequencing analysis of the amplified kDNA was used to investigate the genetic variations among L. major isolates and to correlate the findings with microscopical features and geographical origins.

Results: The quantified amastigote density in the 40 positive touch smears was blindly classified by 3 observers. Grade I, II, III and IV had 7, 13, 9 and 11 cases, respectively. The microscopical features, the mean percentage of neutrophils, lymphocytes and other inflammatory cells, and the leishmanial density of the grades and negative cytopathological samples were contrasting. kDNA amplification of L. major was detected from the cutaneous lesion, and 21 of these amplicons were successfully sequenced.

Conclusions: These results indicate that L. major strains causing CL in southern Iran are genetically diverse; furthermore, a correlation between the genetic heterogeneity of the parasite, the microscopical manifestation and the geographical regions of the disease in humans was found.
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http://dx.doi.org/10.1159/000445865DOI Listing
January 2017

Population-Based Seroprevalence of Malaria in Hormozgan Province, Southeastern Iran: A Low Transmission Area.

Malar Res Treat 2015 12;2015:174570. Epub 2015 Oct 12.

Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran ; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

The seroepidemiological condition of malaria in three main districts of Hormozgan Province, a low transmission area in southeast of Iran, was investigated. Methods. Sera samples (803) were collected from healthy volunteers from the three main districts (Bandar Lengeh in the west, Bandar Abbas in the center, and Bandar Jask in the east) of Hormozgan Province. A questionnaire was used to record the sociodemographic features of the participants during sample collecting. An in-house ELISA test, using crude antigens obtained from cell culture of Plasmodium falciparum, was adapted and used to detect anti-malaria antibodies in the sera. Results. The overall seroprevalence of malaria was 8.7% (70 out of 803 samples). A significant correlation was found between seropositivity and place of residence, where the highest rate of seropositivity was seen in Bandar Lengeh (west of the province). The highest seroprevalence of malaria (13.2%) was seen in the age group of 11-20 years and also in low educated individuals. Correlation between seropositivity and gender, age, and educational levels of the participants was statistically significant (P < 0.05). Conclusion. Findings of this study indicate that the rate of seropositivity to malaria in this area is not high and this might be linked to the success of malaria control programs during the last decades in the region.
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http://dx.doi.org/10.1155/2015/174570DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4620240PMC
November 2015

Leishmania infantum FML pulsed-dendritic cells induce a protective immune response in murine visceral leishmaniasis.

Immunotherapy 2015 ;7(1):3-12

Departmant of Parasitology & Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Aim: To investigate the efficacy of FML loaded dendritic cells (DCs) in protection against visceral leishmaniasis.

Materials & Methods: Mice were immunized with FML- or soluble Leishmania antigen-loaded DCs as well as FML or soluble Leishmania antigen in saponin and challenged with parasite. The levels of cytokines before and after challenge were detected by ELISA. Parasite burden (total Leishman-Donovan unit) was determined after parasite challenge.

Results: FML-saponin induced the highest IFN-γ/IL-4 ratio among vaccinated groups, though this ratio was higher in FML-loaded DCs group subsequent to challenge with Leishmania infantum. Moreover, the greatest reduction in parasite number was detected in mice vaccinated with FML-loaded DCs compared with phosphate-buffered saline-treated mice (p = 0.002).

Conclusion: FML-loaded DCs are one of the promising tools for protection against murine visceral leishmaniasis.
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http://dx.doi.org/10.2217/imt.14.102DOI Listing
September 2015

First detection of acalculous cholecystitis associated with Sarcocystis infection in a patient with AIDS.

Acta Parasitol 2014 Jun 15;59(2):310-5. Epub 2014 May 15.

Department of Microbiology, Fasa University of Medical Sciences, Fasa, Iran.

Acalculous cholecystitis and cholangitis are increasingly being recognized as complications of AIDS. The opportunistic parasites that have been most commonly associated with these disorders are Cryptosporidium species, Isospora belli, Cyclospora cayetanensis and Enterocytozoon bieneusi. The authors performed a parasitological survey on the gallbladder tissue sections of patients underwent cholecystectomy due to chronic acalculous cholecystitis at the Shiraz University of Medical Sciences, Iran. Light microscopic investigation in more than three hundred archived histopathological slides revealed the presence of sexual stages (i.e., mature sporocysts) of a coccidial protozoan in a patient with AIDS who developed acalculous cholecystitis as confirmed by histological, parasitological and molecular tests in which Sarcocystis species was the only identifiable pathogen in gallbladder sections. In the best of our knowledge it's the first documented case of chronic non-calculous cholecystitis due to Sarcocystis parasite in an Iranian AIDS patient from worldwide.
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http://dx.doi.org/10.2478/s11686-014-0243-1DOI Listing
June 2014

Comparison of conventional, molecular, and immunohistochemical methods in diagnosis of typical and atypical cutaneous leishmaniasis.

Arch Pathol Lab Med 2014 Feb;138(2):235-40

From the Brain and Spinal Injury Research Center, Tehran University of Medical Sciences, Iran (Dr Shirian); the Department of Pathology, School of Veterinary Medicine, Shiraz University, Iran (Drs Shirian, Oryan, and Panahi); Basic Sciences in Infectious Diseases, Research Center, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran (Dr Hatam); and the Department of Cytopathology, Dr Daneshbod Pathology Laboratory, Shiraz, Iran (Dr Daneshbod).

Context: Localized cutaneous leishmaniasis (CL) typically presents as papules, crusted nodules, plaques, or noduloulcerative lesions. Atypical CL does not show these features or mimic malignant lesion. In atypical forms, CL may be overlooked because of its similarity to other dermal diseases.

Objective: To compare conventional, molecular, and immunohistochemical methods in the diagnosis of typical and atypical CL.

Design: The kinetoplast DNA, nested, polymerase chain reaction assay and immunohistochemical methods were compared and validated against conventional methods, including cytology and pathology, using 100 specimens of typical and atypical lesions of suspected CL.

Results: Compared with other methods, polymerase chain reaction of the kinetoplast DNA showed the highest sensitivity (typical positive, 100%, 67 of 67; atypical positive, 94%, 31 of 33) and specificity (100%), followed by immunohistochemistry (typical positive, 97%, 65 of 67, with 100% specificity; atypical positives, 94%, 31 of 33, with 100% specificity), and cytology (typical positive, 79%, 53 of 67, with 100% specificity; atypical positive, 58%, 19 of 33, with 100% specificity), followed by pathology (typical positive, 70%, 47 of 67, with 100% specificity; atypical positive, 42%, 14 of 33, with 100% specificity). In addition, polymerase chain reaction enabled identification of 98% (98 of 100) of the positive samples that included strains of Leishmania major (99% [99 of 100] cases) and Leishmania tropica (1% [1 of 100] cases).

Conclusions: Because cytology is cheap and easy to perform with high sensitivity, it is the preferred, primary approach for typical CL, but cytology and pathology do not have sufficient sensitivity for diagnosis of atypical CL cases. Nested polymerase chain reaction and immunohistochemistry are sensitive tests for diagnosis of both typical and atypical CL and are recommended as complementary tests in suspected CL with negative conventional microscopy results.
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http://dx.doi.org/10.5858/arpa.2013-0098-OADOI Listing
February 2014

Three Leishmania/L. species--L. infantum, L. major, L. tropica--as causative agents of mucosal leishmaniasis in Iran.

Pathog Glob Health 2013 Jul;107(5):267-72

Brain and Spinal Cord Injury Research Center, Tehran University of Medical Sciences, Tehran, Iran.

Cases of human oro-mucosal leishmaniasis are mainly reported in areas where Leishmania (Viannia) braziliensis perpetuates and the damages are mainly located at the cartilaginous nasal septum and frontal portions of the nasal fossa. In Iran, an area free of any L.(V) braziliensis, three Leishmania species are known to perpetuate through distinct (i) blood-feeding sand flies and (ii) rodents or (iii) canidae. Thus while establishing the diagnosis of any human oro-mucosal lesions, three Leishmania species - L. infantum, L. major, and L. tropica - must be considered as potential etiological agents of these damages. With these objectives in mind, features such as localization, extent, severity of oro-mucosal lesions, and duration of symptoms at the time of diagnosis were recorded from 11 patients with respect to the presence or absence of cutaneous lesions in other body parts. The biopsy samples were collected from the oro-mucosal and cutaneous lesions and were processed for further identification of the Leishmania species. The lesions ranged from mucosal nodules without ulceration, nodules with erosion, and shallow to deep ulcerations. Leishmania major was isolated from six (55%) cases showing lesions or scars. The scars were restricted to upper and lower extremities. For the other five patients who did not display any signs of former or active cutaneous leishmaniasis, L. major, L. tropica, and L. infantum were isolated from their lesions. In conclusion L. major, L. infantum, and L. tropica, regardless of common tropism, can be seen in mucosal tissues. However, L. major was the predominant species detected from the lesions in the nasal, gingival, and hard and soft palates, and L. tropica was isolated from the gingival and lower lip lesions. Leishmania infantum was isolated from two severe cases of deep mucosal damage displayed by the epiglottis, cricoarytenoid muscle, and laryngeal mucosa. One important finding was the association of L. major with active or scarred skin lesions.
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http://dx.doi.org/10.1179/2047773213Y.0000000098DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4001456PMC
July 2013

Genetic diversity of Leishmania major strains isolated from different clinical forms of cutaneous leishmaniasis in southern Iran based on minicircle kDNA.

Infect Genet Evol 2013 Oct 25;19:226-31. Epub 2013 Jul 25.

Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

Cutaneous leishmaniasis (CL) has been reported from 20 of 31 provinces of Iran. It is a polymorphic disease, which may show various clinical manifestations. Although genetic diversity of the parasite is suggested to be one of the factors influencing the clinical manifestations in CL, there is still no data regarding the genetic variation of Leishmania major based on the sequencing of kDNA .The amplification of the kinetoplast DNA based on 21 isolates of L. major from different clinical forms of CL was carried out using nested-PCR and subsequent sequencing. The clinical presentation was basically of two types: (a) typical lesions and (b) atypical, including erythematous volcanic ulcer, multi infections, lupoid, diffuse, eczematous, verrucous, dry, and nodular lesions. Sequence analysis of the amplified kDNA was used to investigate the genetic variations among L. major isolates and correlate the findings with clinical and histological features. Leishmanial DNA was detected in 98 of 100 cases. L. major and Leishmaniatropica were detected in 97 cases and 1 case, respectively. The Sequence analysis of kDNA from 21 L. major strains showed a high genetic polymorphism of L. major causing CL in southern Iran and correlations among the geographical origin and the clinical manifestations of the disease.
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http://dx.doi.org/10.1016/j.meegid.2013.07.021DOI Listing
October 2013

Molecular, cytological, and immunocytochemical study and kDNA sequencing of laryngeal Leishmania infantum infection.

Parasitol Res 2013 Apr 24;112(4):1799-804. Epub 2012 Dec 24.

Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

Mucosal leishmaniasis is a well-known clinical manifestation of infections mainly caused by New World Leishmania species, especially Leishmania braziliensis (Viannia) in Central and South America. It is extremely uncommon in the world, even in the endemic areas such as Fars Province, Southern Iran. Two male immunocompetent subjects who developed Leishmania mucosal lesion mimicking a laryngeal tumor presented with a several-months history of dysphonia, dyspnea, hoarseness, and odynophagia. Multiple smears from the lesions showed structures resembling the amastigote form of Leishmania. Nested PCR analysis to amplifying a fragment of Leishmania infantum kinetoplastid DNA from the Giemsa-stained smear resulted in a fragment of 680 bp. Sequence analysis of one of the strains showed 98% similarity to L. infantum strain IranJWinf (GenBank accession no. AB678348.1) and 96% similarity to L. infantum isolate MCAN/ES/98/10445 (GenBank accession no. EU437407.1), while another strain showed 97% similarity with two L. infantum strains from kala-azar patient (GenBank accession nos. AJ223725.1 and AF027577.1). Immunocytochemical staining with anti-L. infantum mAb (D2) was positive. Primary mucosal leishmaniasis (ML) may occur in the immunocompetent patients who reside in or travel to endemic areas of leishmaniasis. Mucosal leishmaniasis contracted in endemic areas, such as Iran, has to be considered in the differential diagnosis of lesions in the other mucosa and may occur in previously healthy persons. Therefore, cytology, PCR, and immunocytochemistry-based methods with anti-Leishmania mAb are helpful in the diagnosis of ML.
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http://dx.doi.org/10.1007/s00436-012-3240-zDOI Listing
April 2013

Isoenzyme and ultrastructural characterization of Leishmania tropica axenic amastigotes and promastigotes.

Parasitol Res 2013 Feb 13;112(2):643-8. Epub 2012 Nov 13.

Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Leishmania tropica is one of the main etiological agents of cutaneous leishmaniasis in Iran. For ultrastructural and isoenzyme study, axenic amastigotes were cultured in a brain-heart infusion medium containing 20 % fetal calf serum, pH 4.5, and incubated at 37 °C in 5 % CO(2). Different stages of L. tropica revealed the same isoenzyme profiles after comparing four enzyme systems including phosphoglucomutase, 6-phosphogluconate dehydrogenase, malate dehydrogenase, and nucleoside hydrolase II. Different isoenzyme patterns for glucose-phosphate isomerase, glucose-6-phosphate dehydrogenase, nucleoside hydrolase I, and malic enzyme enzymic systems were seen; thus, these isoenzyme systems among the eight systems studied were more efficient in characterizing L. tropica amastigotes. The structure of the axenic amastigotes was essentially similar to that of the promastigotes except for some important characteristics including the flagellum, flagellar pocket, paraxial rod, and the subpellicular microtubules.
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http://dx.doi.org/10.1007/s00436-012-3179-0DOI Listing
February 2013

Microsporidia and coccidia as causes of persistence diarrhea among liver transplant children: incidence rate and species/genotypes.

Pediatr Infect Dis J 2013 Feb;32(2):185-7

Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

We determined species/genotype(s) of enteric microsporidia and coccidia causing diarrhea among 44 liver transplant children in Shiraz Nemazee hospital using acid-fast-trichrome staining and polymerase chain reaction-sequencing techniques. Enterocytozoon bieneusi (genotype D), Cryptosporidium (parvum and meleagridis) were detected in 6.81% and 11.36% of the children, respectively.
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http://dx.doi.org/10.1097/INF.0b013e318273d95fDOI Listing
February 2013

Mixed mucosal leishmaniasis infection caused by Leishmania tropica and Leishmania major.

J Clin Microbiol 2012 Nov 12;50(11):3805-8. Epub 2012 Sep 12.

Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

Mixed infections with different Leishmania species could explain differences in the clinical courses of these infections. On identification of Leishmania parasites from Iranian patients with mucosal leishmaniasis (ML), a patient with both oral and nasal lesions was found to be concomitantly infected with Leishmania tropica and L. major. Mixed infection was identified by PCR amplification of Leishmania kinetoplast DNA on scraping of cytological smears and histopathological sections. L. major and L. tropica were isolated from the nasal and oral lesions, respectively. These species were also confirmed by immunohistochemistry. This seems to be the first reported case of concurrent ML infection with two Leishmania species. It indicates that, at least in this patient, previous infection with one of these Leishmania species did not protect against infection with the other. This result has important implications for the development of vaccines against leishmaniases and implies careful attention in the treatment of this infectious disease.
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http://dx.doi.org/10.1128/JCM.01469-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486260PMC
November 2012