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.
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.
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.
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.
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.
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. ...
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.
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.
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.
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.
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.
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.
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. ...
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.
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