bahram kazemi | Iran University of Medical Sciences (original) (raw)

Papers by bahram kazemi

Neuroscience Research, 2005

There are some reports regarding the inhibitory effect of pain on tolerance development to analge... more There are some reports regarding the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic formalin induced pain is able to reverse analgesic tolerance to morphine and to evaluate the expression of G(alpha i/o) and G(beta) subunits of G proteins in the context of chronic pain, development of morphine tolerance and their combination. Morphine tolerance was induced by chronic systemic (intraperitoneally, i.p.) or spinal (intrathecally, i.t.) administration of morphine to male Wistar rats weighing 200-240 g and analgesia was assessed using tail flick test. Chronic pain was induced by 4 daily intraplantar injections of 50 microl of 5% formalin. Lumbar spinal tissues were assayed for the expression of G(alpha i/o) and G(beta) proteins using "semiquantitative PCR" normalized to beta-actin gene expression. Results showed that chronic formalin induced pain could reduce and reverse the development of tolerance in rats that had received chronic (i.p. or i.t.) administration of morphine. Chronic administration of morphine did not change G(alpha i/o) gene expression, while chronic pain significantly increased its expression. The expression of G(beta), however, was increased after the chronic administration of morphine, but did not change after the induction of chronic pain. None of these increases were observed when morphine and formalin were administered at the same time. Due to synchronous development of morphine tolerance and changes in expression of G(beta), it may be concluded that the development of tolerance to analgesic effect of morphine is partially mediated by increase in G(beta) gene expression. The increase in G(alpha i/o) genes expression produced by chronic pain may facilitate the opioid signaling pathway and compensate for morphine-induced tolerance.

Brain Research, 2006

It is previously reported that the HPA axis plays role in the inhibitory effect of pain on tolera... more It is previously reported that the HPA axis plays role in the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic co-administration of dexamethasone as a glucocorticoid is also able to prevent or reverse analgesic tolerance to morphine and to compare the expression of G αi/o and G β subunits of G proteins in the context of chronic dexamethasone, development of morphine tolerance and their combination. Analgesic tolerance to morphine was induced by chronic intraperitoneally (i.p.) administration of morphine 20 mg/kg to male Wistar rats weighing 200-240 g within 4 consecutive days and analgesia was assessed using tail-flick test. Chronic dexamethasone was applied using 4 daily i.p.

Scandinavian Journal of Immunology, 2007

Abstract Antinociceptive potency of opioids is greater against various noxious stimuli in animals... more Abstract Antinociceptive potency of opioids is greater against various noxious stimuli in animals with peripheral inflammation. Opioid agonists stimulate activation of G-protein-coupled receptor. Changes in the resting levels of G-protein subtypes could have an effect on intracellular signalling pathways. The present study was designed to investigate the effects of analgesic morphine treatment on the level G-protein subunits mRNA in the presence and absence of inflammation. Our results showed that the carrageenan administration increased G-protein subunits. Administration of analgesic dose of morphine alone and in the presence of inflammation induced different alterations in the levels of G-protein mRNA. Taken together, the results obtained using real time RT-PCR suggested that G-protein genes expression levels following the acute administration of morphine between animals with and without inflammation could influence, at least in part, analgesic responsiveness.

Neuropeptides, 2007

Locus coeruleus (LC) plays a key role in opioid dependence and withdrawal. Chronic morphine admin... more Locus coeruleus (LC) plays a key role in opioid dependence and withdrawal. Chronic morphine administration induces neurochemical adaptations in the noradrenergic system. The nature of signal responsible for opiate-induced adaptations of noradrenergic neurons in LC is not well defined. Neurotrophins-signaling pathways such as brain derived neurotrophic factor (BDNF) and Neurotrophin-3 (NT-3) play a key role for regulating the noradrenergic response of LC neurons to opiates. The nucleus paragigantocellularis (PGi) is one of the two major afferents to LC. The present study was designed to evaluate the expression of BDNF and NT-3 in the context of opiate dependence and withdrawal in PGi. Such data are important because they could reveal the role of PGi as an additional source of BDNF and NT-3 in the neurochemical plasticity of LC neurons. Opiate dependence was induced by a progressive intraperitoneal treatment of morphine. In morphine dependent group PGi nucleus was extracted for gene expression assay 6 h after the last injection of morphine. In spontaneous withdrawal, rats received the same chronic treatment as morphine group. PGi was extracted for gene expression assay 24, 48 and 72 h after the last injection of morphine. PGi nucleus was assayed for the expression of BDNF and NT-3 using semi-quantitative RT-PCR normalized to b-actin gene expression. Results showed that chronic administration of morphine significantly increased BDNF and NT-3 gene expression in PGi. In spontaneous withdrawal, BDNF/NT-3 genes expression were high in comparison to control group. It seems that BDNF/NT-3 -signaling pathway originating from PGi is essential for opiate-induced adaptations of the LC neurons.

Acta Tropica, 2009

Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmanias... more Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmaniasis (VL) caused by Leishmania infantum in the Mediterranean region. The objective of this study was to determine the potential of asymptomatic infected dogs compared with symptomatic ones as a source of L. infantum infection to golden hamster.

Parasitology Research, 2008

The aims of this study are to identify Leishmania species and compare and validate internal trans... more The aims of this study are to identify Leishmania species and compare and validate internal transcribed spacers (ITS) polymerase chain reaction (PCR) assay against parasitological methods for diagnosing cutaneous leishmaniasis (CL). We used the ITS-PCR, parasite culture, and microscopic evaluation of stained smears on 155 specimens from suspected cases of (CL) patients who referred to Mashhad Health Centers (northeast Iran). The PCR indicated the sensitivity (98.8%), correctly diagnosing 86 of the 87 confirmed positive specimens. Microscopy and parasite culture alone showed 79.3% sensitivity (69/87 positive) and 86.2% sensitivity (75/87 positive), respectively, while microscopy and culture in combination improved sensitivity totally to 100% (87/87). The results also revealed that Leishmania tropica species is dominant (96.5%) in the studied regions. This study suggests that both the parasitological techniques reliably were used for the diagnosis of CL, and the ITS-PCR assay without using RFLP analysis is useful for identifying Leishmania species in the area.

This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis... more This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis patients and characterized them by RAPD-PCR technique. Eighty-seven Leishmania isolates from 112 samples were collected from cutaneous leishmaniasis (CL) patients who referred to Mashhad Health Centers from August 2002 to May 2004. Desirable samples (87 isolates) were characterized by RAPD-PCR method using four selected oligoprimers. Electrophoresis patterns from each isolate were compared with reference strains of L. major, L. tropica and L. infantum. The results showed that 94.2% and 5.8% of isolates were similar to L.tropica and L.major reference strain, respectively. Four isolates that were determined by RAPD-PCR as L.major, could produce ulcer at the base tail of BALB/c mice, 4 -12 weeks after inoculation but none of L. tropica isolates produced any lesions at the site of injection in the animals. The results indicate that L. tropica species are dominant in the studied areas of Mashhad city and RAPD-PCR technique is a suitable tool for Leishmania characterization in epidemiological studies.

Patients with diabetes mellitus type II suffer from hyperglycemia because they are not able to us... more Patients with diabetes mellitus type II suffer from hyperglycemia because they are not able to use the insulin that they produce, often due to inadequate function of insulin receptors. There are some evidences that this deficiency is inherited in a dominant autosomal manner and leads to the malfunction of the pancreatic beta cells resulting in insulin excretion disorders. In this study, we sought to identify mutations in the insulin receptor (INSR) gene, which can cause insulin resistance in type II diabetic patients. DNA was extracted from peripheral blood cells of the patients (n = 128) diagnosed with type II diabetes. All 22 exons of the INSR gene of the patients were analyzed for mutations running PCR, conformation-sensitive gel electrophoresis and DNA sequencing, consecutively. Approximately 26% of the patients had genetic mutations; however, most of them were not reported. These mutations include exon 2 (His171Asn, Ile172Ser, Cys196Ser and Ser210Arg), exon 3 (Gly227Asp and Gly232Ser), exon 8 (Thr543Ser), exon 9 (a heterozygote was observed with no change in phenylalanine at position 669), exon 13 (two heterozygotes: Arg890Pro with Asn865 remaining unchanged), exon 14 (Ala906Gly and Pro918Trp with Arg902 unchanged), exon 17 (Val1086Glu) and exon 19 (His1157Gln with Thr1172 unchanged). The lack of similar mutation records in literature and genetic data banks may suggest a geographic pattern for these INSR gene variants in our population.

Veterinary Parasitology, 2009

Cryptosporidium spp. cause enteric diseases in a wide range of animals, including dairy cattle. H... more Cryptosporidium spp. cause enteric diseases in a wide range of animals, including dairy cattle. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle in Gansu province and Ningxia Hui Autonomous Region (NXHAR), northwest China. A total of 2945 dairy feces samples (1257 from Gansu province and 1688 from NXHAR) were collected between December 2012 and March 2014 and were tested by PCR amplification of the small subunit (SSU) rRNA gene. A total of 150 (5.09 %, 58 from Gansu and 92 from NXHAR) samples were PCR-positive for Cryptosporidium, and the prevalence is associated with the region and age of dairy cattle. Species identification showed Cryptosporidium andersoni in 36 samples (24.00 %, 19 from NXHAR and 17 from Gansu), Cryptosporidium ryanae in 24 samples (16.00 %, 13 from NXHAR and 11 from Gansu), Cryptosporidium bovis in 70 samples (46.67 %, 41 from NXHAR and 29 from Gansu), and Cryptosporidium parvum in 20 samples (13.33 %, 19 from NXHAR and 1 from Gansu). A DNA sequence analysis of the gp60 gene suggested that all the 20 C. parvum isolates represented subtype IIdA15G1. These findings indicated the presence of zoonotic Cryptosporidium in Gansu and NXHAR. This is the first report of four species of Cryptosporidium (C. andersoni, C. ryanae, C. bovis, and C. parvum) infection in dairy cattle in Gansu province. This is also the first report of C. ryanae infection in dairy cattle in NXHAR. Effective control strategies should be implemented to prevent and control Cryptosporidium infection in dairy cattle and humans.

Pakistan Journal of Biological Sciences, 2008

ABSTRACT The aim of this study is designing a diagnostic kit for white spot syndrome virus. We de... more ABSTRACT The aim of this study is designing a diagnostic kit for white spot syndrome virus. We designed 2 series of primers for diagnosis of viral VP24 gene and also primers as internal controls which amplify part of genome in both positive and negative samples. DNA of shrimps were extracted and PCR amplification carried out. In this research, a diagnosis kit for white spot disease of shrimps designed and tested using 32 shrimp samples which were dubious to have this disease. White spot virus were found in 23 samples and the other 9 were negative. For extra confirmation, the PCR product was sequenced and deposited to GenBank. We designed a diagnosis kit for white spot disease of shrimps and tested successfully.

Acta Tropica, 2009

This study has identified and characterized the structure of locus D-A, a noncoding short tandem ... more This study has identified and characterized the structure of locus D-A, a noncoding short tandem repeat (STR) region, also known as locus 1-2, in Iranian Entamoeba dispar isolates. This polymorphic locus has been shown to be potentially useful in investigating the molecular epidemiology of Entamoeba histolytica and E. dispar. The genetic polymorphisms in locus D-A in 28 isolates of E. dispar from three different geographic regions of Iran were distinguished using PCR and sequencing, and the results were compared with the E. dispar gene sequences available in GenBank. In all microscopy-positive E. histolytica/E. dispar samples, PCR with species-specific primers was used to amplify a 477-531 bp product, identifying the samples that had E. dispar. Analysis of the sequences revealed a remarkable degree of genetic diversity with regard to size, number and organization of the repeat units among the E. dispar isolates. The sequenced products showed 12 novel E. dispar genotypes, which have been submitted to the GenBank/EMBL/DDBJ database under accession numbers AB354125-AB354136.

Transactions of The Royal Society of Tropical Medicine and Hygiene, 2009

We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestina... more We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestinal complaints in medical centers in Zahedan, Iran. A total of 1562 stool samples was examined from July 2004 to January 2006 using microscopy (direct smear, formalin-ether concentration), xenic culture and PCR techniques. Four hundred and twenty-seven (27.3%) of the patients were infected with one or more intestinal parasites. Giardia lamblia (10.1%), Entamoeba coli (10%), E. hartmanni (1.7%), Blastocystis hominis (2.2%), Chilomastix mesnili (1.7%), Trichomonas hominis (0.7%), E. histolytica/E. dispar (0.51%) and Iodamoeba butschlii (0.45%) were the most prevalent protozoa detected with microscopy. Of the eight microscopy-positive E. histolytica/E. dispar samples, six were identified as E. dispar by PCR/gel electrophoresis, whereas E. histolytica was not detected at all. Although Zahedan is an area with poor hygiene located in a tropical area near the border of Pakistan and Afghanistan, the prevalence of E. histolytica and E. dispar here compared with other parasites and infectious diseases is unexpectedly low.

... In: Methods in Molecular Biology. Vol. 1. Proteins. Edited by John M w.1984b. P:63-75. ... Ac... more ... In: Methods in Molecular Biology. Vol. 1. Proteins. Edited by John M w.1984b. P:63-75. ... Acta Trop. 1996; 61:307-14. Ortona E, Rigana R, Margutti P, Siracusano A. Native and recombinant antigens in the immunodiagnosis of human cystic echinococcosis. Parasite Immunology. ...

Veterinary Parasitology, 2009

Parasitology Research, 2010

Echinococcus granulosus causes human cystic echinococcosis as an important public health problem ... more Echinococcus granulosus causes human cystic echinococcosis as an important public health problem in many regions of the world. There are some problems in primary diagnosis such as cross-reaction with sera from patients with other parasitic disease in serological tests. The use of an appropriate source of antigenic material is a very important and crucial point in the improvement of the serodiagnostic features such as enzyme-linked immunosorbent assay (ELISA) method. We expressed and purified recombinant AgB of Echinococcus granulosus and used as antigen in ELISA method. Serum samples were given from 36 cystic hydatid disease patients that have been confirmed by surgical operation as well as 36 healthy individuals sera were tested by ELISA method using recombinant AgB and compared with commercial kit (Euroimmun) for specificity and sensitivities value. The sensitivity of 91.66% and specificity of 97.22% were determined by homemade kit.

Background: Although a simple and direct method does not exist for the detection of chlamydial in... more Background: Although a simple and direct method does not exist for the detection of chlamydial infections, there are situations in which reliable serological tests, with sensitivity related to a specific antigen, could be helpful. Objective: The aim of this study was to clone the first 1100 bp of the C. trachomatis outer membrane protein 2 (omp2) gene in order to prepare a recombinant protein for use in an ELISA system designed to recognize the anti-C. trachomatis antibody in patient sera. Methods: The PCR product of the chlamydial omp2 gene was cloned in pBluescript and its first 1100 bp was subcloned in the pQE-30 expression vector and induced by IPTG. The recombinant protein was purified by affinity chromatography and its purity was confirmed by SDS-PAGE, gel diffusion and western blot analyses. The purified protein was coated onto a polystyrene microplate and tested by ELISA using patient serum. Results: We have cloned, over-expressed and purified biologically functional recombinant truncated Omp2 from C. trachomatis for use, as a species-specific recognition antigen, in an ELISA system. In this study we determined a cut-off value of 0.345 for this ELISA system using 55 negative sera and measured six positive sera at dilutions of 1:20-1:2560. Conclusion: As a species-specific recognition antigen, the over-expressed and purified recombinant truncated Omp2 from C. trachomatis performed well in an ELISA system.

Neuroscience Research, 2005

There are some reports regarding the inhibitory effect of pain on tolerance development to analge... more There are some reports regarding the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic formalin induced pain is able to reverse analgesic tolerance to morphine and to evaluate the expression of G(alpha i/o) and G(beta) subunits of G proteins in the context of chronic pain, development of morphine tolerance and their combination. Morphine tolerance was induced by chronic systemic (intraperitoneally, i.p.) or spinal (intrathecally, i.t.) administration of morphine to male Wistar rats weighing 200-240 g and analgesia was assessed using tail flick test. Chronic pain was induced by 4 daily intraplantar injections of 50 microl of 5% formalin. Lumbar spinal tissues were assayed for the expression of G(alpha i/o) and G(beta) proteins using "semiquantitative PCR" normalized to beta-actin gene expression. Results showed that chronic formalin induced pain could reduce and reverse the development of tolerance in rats that had received chronic (i.p. or i.t.) administration of morphine. Chronic administration of morphine did not change G(alpha i/o) gene expression, while chronic pain significantly increased its expression. The expression of G(beta), however, was increased after the chronic administration of morphine, but did not change after the induction of chronic pain. None of these increases were observed when morphine and formalin were administered at the same time. Due to synchronous development of morphine tolerance and changes in expression of G(beta), it may be concluded that the development of tolerance to analgesic effect of morphine is partially mediated by increase in G(beta) gene expression. The increase in G(alpha i/o) genes expression produced by chronic pain may facilitate the opioid signaling pathway and compensate for morphine-induced tolerance.

Brain Research, 2006

It is previously reported that the HPA axis plays role in the inhibitory effect of pain on tolera... more It is previously reported that the HPA axis plays role in the inhibitory effect of pain on tolerance development to analgesic effect of opioids. The present study was designed to investigate whether the chronic co-administration of dexamethasone as a glucocorticoid is also able to prevent or reverse analgesic tolerance to morphine and to compare the expression of G αi/o and G β subunits of G proteins in the context of chronic dexamethasone, development of morphine tolerance and their combination. Analgesic tolerance to morphine was induced by chronic intraperitoneally (i.p.) administration of morphine 20 mg/kg to male Wistar rats weighing 200-240 g within 4 consecutive days and analgesia was assessed using tail-flick test. Chronic dexamethasone was applied using 4 daily i.p.

Scandinavian Journal of Immunology, 2007

Abstract Antinociceptive potency of opioids is greater against various noxious stimuli in animals... more Abstract Antinociceptive potency of opioids is greater against various noxious stimuli in animals with peripheral inflammation. Opioid agonists stimulate activation of G-protein-coupled receptor. Changes in the resting levels of G-protein subtypes could have an effect on intracellular signalling pathways. The present study was designed to investigate the effects of analgesic morphine treatment on the level G-protein subunits mRNA in the presence and absence of inflammation. Our results showed that the carrageenan administration increased G-protein subunits. Administration of analgesic dose of morphine alone and in the presence of inflammation induced different alterations in the levels of G-protein mRNA. Taken together, the results obtained using real time RT-PCR suggested that G-protein genes expression levels following the acute administration of morphine between animals with and without inflammation could influence, at least in part, analgesic responsiveness.

Neuropeptides, 2007

Locus coeruleus (LC) plays a key role in opioid dependence and withdrawal. Chronic morphine admin... more Locus coeruleus (LC) plays a key role in opioid dependence and withdrawal. Chronic morphine administration induces neurochemical adaptations in the noradrenergic system. The nature of signal responsible for opiate-induced adaptations of noradrenergic neurons in LC is not well defined. Neurotrophins-signaling pathways such as brain derived neurotrophic factor (BDNF) and Neurotrophin-3 (NT-3) play a key role for regulating the noradrenergic response of LC neurons to opiates. The nucleus paragigantocellularis (PGi) is one of the two major afferents to LC. The present study was designed to evaluate the expression of BDNF and NT-3 in the context of opiate dependence and withdrawal in PGi. Such data are important because they could reveal the role of PGi as an additional source of BDNF and NT-3 in the neurochemical plasticity of LC neurons. Opiate dependence was induced by a progressive intraperitoneal treatment of morphine. In morphine dependent group PGi nucleus was extracted for gene expression assay 6 h after the last injection of morphine. In spontaneous withdrawal, rats received the same chronic treatment as morphine group. PGi was extracted for gene expression assay 24, 48 and 72 h after the last injection of morphine. PGi nucleus was assayed for the expression of BDNF and NT-3 using semi-quantitative RT-PCR normalized to b-actin gene expression. Results showed that chronic administration of morphine significantly increased BDNF and NT-3 gene expression in PGi. In spontaneous withdrawal, BDNF/NT-3 genes expression were high in comparison to control group. It seems that BDNF/NT-3 -signaling pathway originating from PGi is essential for opiate-induced adaptations of the LC neurons.

Acta Tropica, 2009

Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmanias... more Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmaniasis (VL) caused by Leishmania infantum in the Mediterranean region. The objective of this study was to determine the potential of asymptomatic infected dogs compared with symptomatic ones as a source of L. infantum infection to golden hamster.

Parasitology Research, 2008

The aims of this study are to identify Leishmania species and compare and validate internal trans... more The aims of this study are to identify Leishmania species and compare and validate internal transcribed spacers (ITS) polymerase chain reaction (PCR) assay against parasitological methods for diagnosing cutaneous leishmaniasis (CL). We used the ITS-PCR, parasite culture, and microscopic evaluation of stained smears on 155 specimens from suspected cases of (CL) patients who referred to Mashhad Health Centers (northeast Iran). The PCR indicated the sensitivity (98.8%), correctly diagnosing 86 of the 87 confirmed positive specimens. Microscopy and parasite culture alone showed 79.3% sensitivity (69/87 positive) and 86.2% sensitivity (75/87 positive), respectively, while microscopy and culture in combination improved sensitivity totally to 100% (87/87). The results also revealed that Leishmania tropica species is dominant (96.5%) in the studied regions. This study suggests that both the parasitological techniques reliably were used for the diagnosis of CL, and the ITS-PCR assay without using RFLP analysis is useful for identifying Leishmania species in the area.

This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis... more This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis patients and characterized them by RAPD-PCR technique. Eighty-seven Leishmania isolates from 112 samples were collected from cutaneous leishmaniasis (CL) patients who referred to Mashhad Health Centers from August 2002 to May 2004. Desirable samples (87 isolates) were characterized by RAPD-PCR method using four selected oligoprimers. Electrophoresis patterns from each isolate were compared with reference strains of L. major, L. tropica and L. infantum. The results showed that 94.2% and 5.8% of isolates were similar to L.tropica and L.major reference strain, respectively. Four isolates that were determined by RAPD-PCR as L.major, could produce ulcer at the base tail of BALB/c mice, 4 -12 weeks after inoculation but none of L. tropica isolates produced any lesions at the site of injection in the animals. The results indicate that L. tropica species are dominant in the studied areas of Mashhad city and RAPD-PCR technique is a suitable tool for Leishmania characterization in epidemiological studies.

Patients with diabetes mellitus type II suffer from hyperglycemia because they are not able to us... more Patients with diabetes mellitus type II suffer from hyperglycemia because they are not able to use the insulin that they produce, often due to inadequate function of insulin receptors. There are some evidences that this deficiency is inherited in a dominant autosomal manner and leads to the malfunction of the pancreatic beta cells resulting in insulin excretion disorders. In this study, we sought to identify mutations in the insulin receptor (INSR) gene, which can cause insulin resistance in type II diabetic patients. DNA was extracted from peripheral blood cells of the patients (n = 128) diagnosed with type II diabetes. All 22 exons of the INSR gene of the patients were analyzed for mutations running PCR, conformation-sensitive gel electrophoresis and DNA sequencing, consecutively. Approximately 26% of the patients had genetic mutations; however, most of them were not reported. These mutations include exon 2 (His171Asn, Ile172Ser, Cys196Ser and Ser210Arg), exon 3 (Gly227Asp and Gly232Ser), exon 8 (Thr543Ser), exon 9 (a heterozygote was observed with no change in phenylalanine at position 669), exon 13 (two heterozygotes: Arg890Pro with Asn865 remaining unchanged), exon 14 (Ala906Gly and Pro918Trp with Arg902 unchanged), exon 17 (Val1086Glu) and exon 19 (His1157Gln with Thr1172 unchanged). The lack of similar mutation records in literature and genetic data banks may suggest a geographic pattern for these INSR gene variants in our population.

Veterinary Parasitology, 2009

Cryptosporidium spp. cause enteric diseases in a wide range of animals, including dairy cattle. H... more Cryptosporidium spp. cause enteric diseases in a wide range of animals, including dairy cattle. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle in Gansu province and Ningxia Hui Autonomous Region (NXHAR), northwest China. A total of 2945 dairy feces samples (1257 from Gansu province and 1688 from NXHAR) were collected between December 2012 and March 2014 and were tested by PCR amplification of the small subunit (SSU) rRNA gene. A total of 150 (5.09 %, 58 from Gansu and 92 from NXHAR) samples were PCR-positive for Cryptosporidium, and the prevalence is associated with the region and age of dairy cattle. Species identification showed Cryptosporidium andersoni in 36 samples (24.00 %, 19 from NXHAR and 17 from Gansu), Cryptosporidium ryanae in 24 samples (16.00 %, 13 from NXHAR and 11 from Gansu), Cryptosporidium bovis in 70 samples (46.67 %, 41 from NXHAR and 29 from Gansu), and Cryptosporidium parvum in 20 samples (13.33 %, 19 from NXHAR and 1 from Gansu). A DNA sequence analysis of the gp60 gene suggested that all the 20 C. parvum isolates represented subtype IIdA15G1. These findings indicated the presence of zoonotic Cryptosporidium in Gansu and NXHAR. This is the first report of four species of Cryptosporidium (C. andersoni, C. ryanae, C. bovis, and C. parvum) infection in dairy cattle in Gansu province. This is also the first report of C. ryanae infection in dairy cattle in NXHAR. Effective control strategies should be implemented to prevent and control Cryptosporidium infection in dairy cattle and humans.

Pakistan Journal of Biological Sciences, 2008

ABSTRACT The aim of this study is designing a diagnostic kit for white spot syndrome virus. We de... more ABSTRACT The aim of this study is designing a diagnostic kit for white spot syndrome virus. We designed 2 series of primers for diagnosis of viral VP24 gene and also primers as internal controls which amplify part of genome in both positive and negative samples. DNA of shrimps were extracted and PCR amplification carried out. In this research, a diagnosis kit for white spot disease of shrimps designed and tested using 32 shrimp samples which were dubious to have this disease. White spot virus were found in 23 samples and the other 9 were negative. For extra confirmation, the PCR product was sequenced and deposited to GenBank. We designed a diagnosis kit for white spot disease of shrimps and tested successfully.

Acta Tropica, 2009

This study has identified and characterized the structure of locus D-A, a noncoding short tandem ... more This study has identified and characterized the structure of locus D-A, a noncoding short tandem repeat (STR) region, also known as locus 1-2, in Iranian Entamoeba dispar isolates. This polymorphic locus has been shown to be potentially useful in investigating the molecular epidemiology of Entamoeba histolytica and E. dispar. The genetic polymorphisms in locus D-A in 28 isolates of E. dispar from three different geographic regions of Iran were distinguished using PCR and sequencing, and the results were compared with the E. dispar gene sequences available in GenBank. In all microscopy-positive E. histolytica/E. dispar samples, PCR with species-specific primers was used to amplify a 477-531 bp product, identifying the samples that had E. dispar. Analysis of the sequences revealed a remarkable degree of genetic diversity with regard to size, number and organization of the repeat units among the E. dispar isolates. The sequenced products showed 12 novel E. dispar genotypes, which have been submitted to the GenBank/EMBL/DDBJ database under accession numbers AB354125-AB354136.

Transactions of The Royal Society of Tropical Medicine and Hygiene, 2009

We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestina... more We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestinal complaints in medical centers in Zahedan, Iran. A total of 1562 stool samples was examined from July 2004 to January 2006 using microscopy (direct smear, formalin-ether concentration), xenic culture and PCR techniques. Four hundred and twenty-seven (27.3%) of the patients were infected with one or more intestinal parasites. Giardia lamblia (10.1%), Entamoeba coli (10%), E. hartmanni (1.7%), Blastocystis hominis (2.2%), Chilomastix mesnili (1.7%), Trichomonas hominis (0.7%), E. histolytica/E. dispar (0.51%) and Iodamoeba butschlii (0.45%) were the most prevalent protozoa detected with microscopy. Of the eight microscopy-positive E. histolytica/E. dispar samples, six were identified as E. dispar by PCR/gel electrophoresis, whereas E. histolytica was not detected at all. Although Zahedan is an area with poor hygiene located in a tropical area near the border of Pakistan and Afghanistan, the prevalence of E. histolytica and E. dispar here compared with other parasites and infectious diseases is unexpectedly low.

... In: Methods in Molecular Biology. Vol. 1. Proteins. Edited by John M w.1984b. P:63-75. ... Ac... more ... In: Methods in Molecular Biology. Vol. 1. Proteins. Edited by John M w.1984b. P:63-75. ... Acta Trop. 1996; 61:307-14. Ortona E, Rigana R, Margutti P, Siracusano A. Native and recombinant antigens in the immunodiagnosis of human cystic echinococcosis. Parasite Immunology. ...

Veterinary Parasitology, 2009

Parasitology Research, 2010

Echinococcus granulosus causes human cystic echinococcosis as an important public health problem ... more Echinococcus granulosus causes human cystic echinococcosis as an important public health problem in many regions of the world. There are some problems in primary diagnosis such as cross-reaction with sera from patients with other parasitic disease in serological tests. The use of an appropriate source of antigenic material is a very important and crucial point in the improvement of the serodiagnostic features such as enzyme-linked immunosorbent assay (ELISA) method. We expressed and purified recombinant AgB of Echinococcus granulosus and used as antigen in ELISA method. Serum samples were given from 36 cystic hydatid disease patients that have been confirmed by surgical operation as well as 36 healthy individuals sera were tested by ELISA method using recombinant AgB and compared with commercial kit (Euroimmun) for specificity and sensitivities value. The sensitivity of 91.66% and specificity of 97.22% were determined by homemade kit.

Background: Although a simple and direct method does not exist for the detection of chlamydial in... more Background: Although a simple and direct method does not exist for the detection of chlamydial infections, there are situations in which reliable serological tests, with sensitivity related to a specific antigen, could be helpful. Objective: The aim of this study was to clone the first 1100 bp of the C. trachomatis outer membrane protein 2 (omp2) gene in order to prepare a recombinant protein for use in an ELISA system designed to recognize the anti-C. trachomatis antibody in patient sera. Methods: The PCR product of the chlamydial omp2 gene was cloned in pBluescript and its first 1100 bp was subcloned in the pQE-30 expression vector and induced by IPTG. The recombinant protein was purified by affinity chromatography and its purity was confirmed by SDS-PAGE, gel diffusion and western blot analyses. The purified protein was coated onto a polystyrene microplate and tested by ELISA using patient serum. Results: We have cloned, over-expressed and purified biologically functional recombinant truncated Omp2 from C. trachomatis for use, as a species-specific recognition antigen, in an ELISA system. In this study we determined a cut-off value of 0.345 for this ELISA system using 55 negative sera and measured six positive sera at dilutions of 1:20-1:2560. Conclusion: As a species-specific recognition antigen, the over-expressed and purified recombinant truncated Omp2 from C. trachomatis performed well in an ELISA system.