Berk Saglam, Bora Ergin, Nazlı Coskun Beyatli, Kaan Arslan, Turgut Bastug, Nuhan Purali
Abstract Objectives Voltage-gated calcium channels are essential elements in development of many cellular processes like electrical signaling, contraction secretion and gene expression. There has been a fair amount of information about the functional and structural properties of the calcium channels in mammalian species. Crayfish serves as a model animal for many types of experiments. However, there has been no information related to the molecular and genetic properties of the calcium channels in the crayfish. Methods Conventional cloning methods, three-dimensional structural calculations, docking experiments have been conducted. Results An mRNA 7,791 bp in size has been cloned. The coding region has been translated into an alpha peptide with 1,942 residues. The cloned protein sequence has similarity to other L-type voltage-gated calcium channel sequences from the neighboring species. Three-dimensional structure, in reference to human L-type voltage-gated calcium channel, has been calculated. Known calcium channel blockers, nifedipine, verapamil and diltiazem have been successfully docked on the calculated three-dimensional model. Conclusions Considering the similarity assay in the National Center for Biotechnology Information (NCBI) platform, the three-dimensional structural calculations and the docking experiments it was concluded that the cloned mRNA codes an alpha peptide for a putative voltage-gated calcium channel protein in the crayfish. In the present work by using the conventional molecular biology methods a complete mRNA coding a putative calcium channel has been de novo cloned. Three-dimensional structure of the related protein has been calculated and several pharmacological agents blocking the channel have been docked to the identified receptor sites.
{"title":"Cloning and <i>in silico</i> investigation of a putative voltage-gated calcium channel gene and protein in <i>Astacus Leptodactylus</i>","authors":"Berk Saglam, Bora Ergin, Nazlı Coskun Beyatli, Kaan Arslan, Turgut Bastug, Nuhan Purali","doi":"10.1515/tjb-2023-0143","DOIUrl":"https://doi.org/10.1515/tjb-2023-0143","url":null,"abstract":"Abstract Objectives Voltage-gated calcium channels are essential elements in development of many cellular processes like electrical signaling, contraction secretion and gene expression. There has been a fair amount of information about the functional and structural properties of the calcium channels in mammalian species. Crayfish serves as a model animal for many types of experiments. However, there has been no information related to the molecular and genetic properties of the calcium channels in the crayfish. Methods Conventional cloning methods, three-dimensional structural calculations, docking experiments have been conducted. Results An mRNA 7,791 bp in size has been cloned. The coding region has been translated into an alpha peptide with 1,942 residues. The cloned protein sequence has similarity to other L-type voltage-gated calcium channel sequences from the neighboring species. Three-dimensional structure, in reference to human L-type voltage-gated calcium channel, has been calculated. Known calcium channel blockers, nifedipine, verapamil and diltiazem have been successfully docked on the calculated three-dimensional model. Conclusions Considering the similarity assay in the National Center for Biotechnology Information (NCBI) platform, the three-dimensional structural calculations and the docking experiments it was concluded that the cloned mRNA codes an alpha peptide for a putative voltage-gated calcium channel protein in the crayfish. In the present work by using the conventional molecular biology methods a complete mRNA coding a putative calcium channel has been de novo cloned. Three-dimensional structure of the related protein has been calculated and several pharmacological agents blocking the channel have been docked to the identified receptor sites.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"8 7","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136227159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kamran Abbasi, Parveen Ali, Virginia Barbour, Thomas Benfield, Kirsten Bibbins-Domingo, Stephen Hancocks, Richard Horton, Laurie Laybourn-Langton, Robert Mash, Peush Sahni, Wadeia Mohammad Sharief, Paul Yonga, Chris Zielinski
{"title":"Time to treat the climate and nature crisis as one indivisible global health emergency","authors":"Kamran Abbasi, Parveen Ali, Virginia Barbour, Thomas Benfield, Kirsten Bibbins-Domingo, Stephen Hancocks, Richard Horton, Laurie Laybourn-Langton, Robert Mash, Peush Sahni, Wadeia Mohammad Sharief, Paul Yonga, Chris Zielinski","doi":"10.1515/tjb-2023-2005","DOIUrl":"https://doi.org/10.1515/tjb-2023-2005","url":null,"abstract":"","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"9 5","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135431361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives The key component of neuroprotection after cerebral ischemia–reperfusion (I–R) injury is mitochondrial improvement. By focusing on the function of mitochondrial biogenesis and ATP-sensitive potassium (mK–ATP) channels and inflammatory responses, the current study assessed the neuroprotective potentials of lemon essential oil, D-limonene (LIM), in rats with cerebral I–R injury. Methods In order to simulate cerebral I–R injury, Sprague Dawley rats (n=72) were subjected to a two h local ischemia induced by middle cerebral artery blockage, followed by a 24 h reperfusion period. Five minutes before starting reperfusion, rats were intraperitoneally given LIM at doses of 10 or 100 mg/kg. Cerebral infarct volume was assessed by triphenyl-tetrazolium chloride staining, brain activity by behavioral tests and mitochondrial function/biogenesis, as well as proinflammatory cytokines by fluorometry, immunoblotting and other related techniques. Results When compared to the untreated control group, the administration of LIM substantially and dose-dependently decreased cerebral infarct volumes and neurological deficits (p<0.01). I–R injury-induced alterations in mitochondrial membrane depolarization, mitochondrial reactive oxygen species (mitoROS), and superoxide dismutase (mnSOD), as well as inflammatory cytokines TNF-α, IL-6 and IL-1β, were all significantly reversed after treatment with LIM 100 mg/kg (p<0.01). Additionally, this dose of LIM increased the expression of mitochondrial biogenesis proteins PGC-1α, TFAM, and NRF1. Interestingly, blockage of mK–ATP channels by 5-hydoxydecanoate diminished the effects of LIM on cerebral positive endpoints, cytokines production, and mitochondrial function/biogenesis. Conclusions Thus, the strong neuroprotective effects of LIM-postconditioning were mediated by an increase in mK–ATP channel activity, which improved mitochondrial biogenesis and suppressed inflammatory responses.
{"title":"Postconditioning with D-limonene exerts neuroprotection in rats via enhancing mitochondrial activity","authors":"Leguo Zhang, Zeyu Zhao, Jianpu Jia, Liran Zhang, Ruixue Xia, Cuimin Zhu","doi":"10.1515/tjb-2022-0290","DOIUrl":"https://doi.org/10.1515/tjb-2022-0290","url":null,"abstract":"Abstract Objectives The key component of neuroprotection after cerebral ischemia–reperfusion (I–R) injury is mitochondrial improvement. By focusing on the function of mitochondrial biogenesis and ATP-sensitive potassium (mK–ATP) channels and inflammatory responses, the current study assessed the neuroprotective potentials of lemon essential oil, D-limonene (LIM), in rats with cerebral I–R injury. Methods In order to simulate cerebral I–R injury, Sprague Dawley rats (n=72) were subjected to a two h local ischemia induced by middle cerebral artery blockage, followed by a 24 h reperfusion period. Five minutes before starting reperfusion, rats were intraperitoneally given LIM at doses of 10 or 100 mg/kg. Cerebral infarct volume was assessed by triphenyl-tetrazolium chloride staining, brain activity by behavioral tests and mitochondrial function/biogenesis, as well as proinflammatory cytokines by fluorometry, immunoblotting and other related techniques. Results When compared to the untreated control group, the administration of LIM substantially and dose-dependently decreased cerebral infarct volumes and neurological deficits (p<0.01). I–R injury-induced alterations in mitochondrial membrane depolarization, mitochondrial reactive oxygen species (mitoROS), and superoxide dismutase (mnSOD), as well as inflammatory cytokines TNF-α, IL-6 and IL-1β, were all significantly reversed after treatment with LIM 100 mg/kg (p<0.01). Additionally, this dose of LIM increased the expression of mitochondrial biogenesis proteins PGC-1α, TFAM, and NRF1. Interestingly, blockage of mK–ATP channels by 5-hydoxydecanoate diminished the effects of LIM on cerebral positive endpoints, cytokines production, and mitochondrial function/biogenesis. Conclusions Thus, the strong neuroprotective effects of LIM-postconditioning were mediated by an increase in mK–ATP channel activity, which improved mitochondrial biogenesis and suppressed inflammatory responses.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"45 5","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136318335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives The aim of this study was to compare the analytical performance of the KIMS (kinetic interaction of microparticles in solution) immunochemical method with a validated in-house and a commercial LC-MS/MS method. Methods The urine samples of the 100 subjects were included in the present study. The urine samples were analysed with Roche DAT immunochemical method based on KIMS method. In-house LC-MS/MS method was validated for 58 parameters according to the CLSI C62-A recommendations with the following parameters: matrix effect, lower limit of quantification (LLOQ), linearity, intra-day and inter-day precision and accuracy. Eureka Lab Division Drugs of Abuse kit was used as the commercial LC-MS/MS method. Results The immunochemical method had a satisfactory performance with specificity, sensitivity and accuracy values above 80 % and met the DRUID recommendation except benzodiazepines. The sensitivity and specificity of the immunochemical method were between 97–100 % and 84–100 %, respectively (except for benzodiazepines). The bias obtained for THC-COOH, morphine and codeine parameters were −17.5, 24.6 and 43.6 between two LC-MS/MS methods. The commercial method had a tendency to have a negative bias except for cannabinoids. Conclusions The analytical performance of the KIMS-based urine immunochemical method was found to be satisfactory for the intended use, except for benzodiazepines. The validated urine in-house LC-MS/MS method was found to be a good alternative for confirmation of substance abuse.
{"title":"Comparison of the immunoassay method with the commercial and in-house LC-MS/MS methods for substance abuse in urine","authors":"Gamze Avcioglu, Gulsen Yilmaz, Safak Yalcin Sahiner, L. Didem Kozaci, Ceylan Bal, Fatma Meric Yilmaz","doi":"10.1515/tjb-2022-0286","DOIUrl":"https://doi.org/10.1515/tjb-2022-0286","url":null,"abstract":"Abstract Objectives The aim of this study was to compare the analytical performance of the KIMS (kinetic interaction of microparticles in solution) immunochemical method with a validated in-house and a commercial LC-MS/MS method. Methods The urine samples of the 100 subjects were included in the present study. The urine samples were analysed with Roche DAT immunochemical method based on KIMS method. In-house LC-MS/MS method was validated for 58 parameters according to the CLSI C62-A recommendations with the following parameters: matrix effect, lower limit of quantification (LLOQ), linearity, intra-day and inter-day precision and accuracy. Eureka Lab Division Drugs of Abuse kit was used as the commercial LC-MS/MS method. Results The immunochemical method had a satisfactory performance with specificity, sensitivity and accuracy values above 80 % and met the DRUID recommendation except benzodiazepines. The sensitivity and specificity of the immunochemical method were between 97–100 % and 84–100 %, respectively (except for benzodiazepines). The bias obtained for THC-COOH, morphine and codeine parameters were −17.5, 24.6 and 43.6 between two LC-MS/MS methods. The commercial method had a tendency to have a negative bias except for cannabinoids. Conclusions The analytical performance of the KIMS-based urine immunochemical method was found to be satisfactory for the intended use, except for benzodiazepines. The validated urine in-house LC-MS/MS method was found to be a good alternative for confirmation of substance abuse.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"10 3","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135565733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cemile Zontul, Ayca Tas, Emrullah Hayta, Yavuz Silig
Abstract Objectives Fibromyalgia syndrome (FMS) is a chronic pain syndrome characterized by widespread body pain over a long period, the cause of which is not yet clearly known. FMS patients usually have high pain sensitivity. We aimed to investigate whether rs4148855 and rs2288646 polymorphisms of acid-sensing ion channel 3 ( ASIC3 ), one of the factors contributing to pain, cause a predisposition to FMS in the Turkish population. Methods ASIC3 gene rs4148855 and rs2288646 polymorphisms in DNA samples obtained from blood samples of 175 patients with FMS and 176 healthy individuals were analyzed by real-time polymerase chain reaction (RT-PCR) using a hydrolysis probe. Statistical data were obtained by chi-square ( χ 2 ) test and logistic regression analysis. Results No significant association was found between ASIC3 gene rs4148855 and rs2288646 polymorphisms and the Turkish population’s FMS group and control group (p>0.05). Conclusions As a result, no significant association was found between the genotype and allele distributions of ASIC3 polymorphism (rs4148855 and rs2288646) in patients with FMS compared to controls in the Turkish population. Further studies are needed to elucidate the relationship between ion channels and FMS to elucidate the mechanisms of FMS.
{"title":"The relationship between <i>ASIC3</i> gene polymorphism and fibromyalgia syndrome","authors":"Cemile Zontul, Ayca Tas, Emrullah Hayta, Yavuz Silig","doi":"10.1515/tjb-2023-0101","DOIUrl":"https://doi.org/10.1515/tjb-2023-0101","url":null,"abstract":"Abstract Objectives Fibromyalgia syndrome (FMS) is a chronic pain syndrome characterized by widespread body pain over a long period, the cause of which is not yet clearly known. FMS patients usually have high pain sensitivity. We aimed to investigate whether rs4148855 and rs2288646 polymorphisms of acid-sensing ion channel 3 ( ASIC3 ), one of the factors contributing to pain, cause a predisposition to FMS in the Turkish population. Methods ASIC3 gene rs4148855 and rs2288646 polymorphisms in DNA samples obtained from blood samples of 175 patients with FMS and 176 healthy individuals were analyzed by real-time polymerase chain reaction (RT-PCR) using a hydrolysis probe. Statistical data were obtained by chi-square ( χ 2 ) test and logistic regression analysis. Results No significant association was found between ASIC3 gene rs4148855 and rs2288646 polymorphisms and the Turkish population’s FMS group and control group (p>0.05). Conclusions As a result, no significant association was found between the genotype and allele distributions of ASIC3 polymorphism (rs4148855 and rs2288646) in patients with FMS compared to controls in the Turkish population. Further studies are needed to elucidate the relationship between ion channels and FMS to elucidate the mechanisms of FMS.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135666887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives A comprehensive understanding of the role of PON enzymatic activities may play an important role in the etiology and prevention of many cancers. PON1 is known as a potent antioxidant that scavenges free radicals in the human body. The enzymatic activities of paraoxonase (PON1) and mitogen-activated protein kinase (MAPK) signalling pathways in colorectal cancer are being investigated to determine whether they hold promise for novel diagnostic or therapeutic applications in colorectal cancer. Methods HT-29 colon cancer cell lines and CCD-18Co colon cell lines were used. B-Raf, p-B-Raf, ERK, and p-ERK proteins involved in MAPK signalling pathways and serum levels of PON1 were detected and analyzed by the Western blotting method. Results The levels and activity of PON1 enzyme were significantly decreased in HT-29 cells compared to CCD-18Co cells (p=0.0173 and p=0.0281, respectively). The levels of p-B-Raf and p-ERK, which activates the MAPK signalling pathway, were significantly increased in HT-29 cells (p=0.0037 and p=0.0074, respectively). Conclusions A positive correlation was found between PON1 level and PON1 enzyme activity (p=0.008). A negative correlation was found between PON1 level and p-B-Raf/p-ERK protein levels, which play a role in the MAPK signalling pathway (p=0.016 and p=0.036, respectively). A negative correlation was also found between PON1 enzyme activity and p-B-Raf/p-ERK protein levels (p=0.060 and p=0.037, respectively). It is suggested that the increase of proteins involved in the MAPK signaling pathway in cancer cells is caused by the decreasing serum levels of PON1 and enzymatic activity of PON1.
摘要目的全面了解PON酶活性的作用可能在许多癌症的病因和预防中发挥重要作用。PON1被认为是一种有效的抗氧化剂，可以清除人体内的自由基。人们正在研究结直肠癌中对氧磷酶(PON1)和丝裂原活化蛋白激酶(MAPK)信号通路的酶活性，以确定它们是否有望在结直肠癌的诊断或治疗中应用。方法采用HT-29结肠癌细胞系和CCD-18Co结肠癌细胞系。采用Western blotting法检测MAPK信号通路相关的B-Raf、p-B-Raf、ERK和p-ERK蛋白及PON1的血清水平。结果HT-29细胞PON1酶表达水平和活性较cd - 18co细胞明显降低(p=0.0173和p=0.0281)。激活MAPK信号通路的p- b - raf和p- erk水平在HT-29细胞中显著升高(p=0.0037和p=0.0074)。结论PON1水平与PON1酶活性呈正相关(p=0.008)。PON1水平与参与MAPK信号通路的p- b - raf /p- erk蛋白水平呈负相关(p=0.016和p=0.036)。PON1酶活性与p- b - raf /p- erk蛋白水平呈负相关(p=0.060和p=0.037)。提示肿瘤细胞中MAPK信号通路相关蛋白的增加是由血清PON1水平和PON1酶活性的降低引起的。
{"title":"The attraction of paraoxonase-1 associated with the MAPK pathway on colon carcinoma cells","authors":"Muhammet Örnek, Özen Özensoy Güler","doi":"10.1515/tjb-2023-0072","DOIUrl":"https://doi.org/10.1515/tjb-2023-0072","url":null,"abstract":"Abstract Objectives A comprehensive understanding of the role of PON enzymatic activities may play an important role in the etiology and prevention of many cancers. PON1 is known as a potent antioxidant that scavenges free radicals in the human body. The enzymatic activities of paraoxonase (PON1) and mitogen-activated protein kinase (MAPK) signalling pathways in colorectal cancer are being investigated to determine whether they hold promise for novel diagnostic or therapeutic applications in colorectal cancer. Methods HT-29 colon cancer cell lines and CCD-18Co colon cell lines were used. B-Raf, p-B-Raf, ERK, and p-ERK proteins involved in MAPK signalling pathways and serum levels of PON1 were detected and analyzed by the Western blotting method. Results The levels and activity of PON1 enzyme were significantly decreased in HT-29 cells compared to CCD-18Co cells (p=0.0173 and p=0.0281, respectively). The levels of p-B-Raf and p-ERK, which activates the MAPK signalling pathway, were significantly increased in HT-29 cells (p=0.0037 and p=0.0074, respectively). Conclusions A positive correlation was found between PON1 level and PON1 enzyme activity (p=0.008). A negative correlation was found between PON1 level and p-B-Raf/p-ERK protein levels, which play a role in the MAPK signalling pathway (p=0.016 and p=0.036, respectively). A negative correlation was also found between PON1 enzyme activity and p-B-Raf/p-ERK protein levels (p=0.060 and p=0.037, respectively). It is suggested that the increase of proteins involved in the MAPK signaling pathway in cancer cells is caused by the decreasing serum levels of PON1 and enzymatic activity of PON1.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"24 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136078246","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jyot Amrita, Amarjit S. Bhanwer, ArvinderPal Singh
Abstract Objectives We aimed to explore the association of obesity and hypertension and further their association with AdipoQ gene polymorphism in North Indian postmenopausal women of Punjab. Methods A total of 523 postmenopausal women (PMW) were enrolled (PMW with CVD=265 and PMW without CVD=258). Anthropometric measurements such as weight, height, hip circumference (HC), waist circumference (WC), waist hip ratio (WHR) and body mass index (BMI) for all the subjects were recorded in accordance to WHO 2000 protocol. For hypertension, guidelines of the Joint National Committee (JNC-VII) of high blood pressure were considered. Genotyping of AdipoQ (G>T) gene polymorphism was done by RFLP-PCR analysis. Results The comparison of the frequency distribution of alleles and genotypes of AdipoQ (+276G>T) gene polymorphism showed a significant distribution (p<0.05) among subjects with and without CVD. The risk for CVD was high (∼9 fold) among carriers of +276T allele towards CVD predisposition. Obese women with CVD under the recessive model conferred ∼8 fold high risk (p=0.001) and +276T allele and TT genotype of non-obese women with CVD for BMI <25 also conferred ∼9 fold high risk. Hypertension also acted as a strong risk factor related to CVD (p=0.0001). Under the recessive model, hypertensive PMW with CVD conferred 7–9 fold higher risk however, normotensive women with CVD also conferred 9∼10-fold risk towards CVD predisposition. Conclusions The T allele carriers of AdipoQ gene is strongly associated with risk factors such as obesity and hypertension pertaining to cardiovascular disease. Early detection of these risk factors may serve as a CVD preventative intervention.
{"title":"Association of AdipoQ (G>T) gene polymorphism with obesity and hypertension in North Indian postmenopausal women of Punjab","authors":"Jyot Amrita, Amarjit S. Bhanwer, ArvinderPal Singh","doi":"10.1515/tjb-2023-0073","DOIUrl":"https://doi.org/10.1515/tjb-2023-0073","url":null,"abstract":"Abstract Objectives We aimed to explore the association of obesity and hypertension and further their association with AdipoQ gene polymorphism in North Indian postmenopausal women of Punjab. Methods A total of 523 postmenopausal women (PMW) were enrolled (PMW with CVD=265 and PMW without CVD=258). Anthropometric measurements such as weight, height, hip circumference (HC), waist circumference (WC), waist hip ratio (WHR) and body mass index (BMI) for all the subjects were recorded in accordance to WHO 2000 protocol. For hypertension, guidelines of the Joint National Committee (JNC-VII) of high blood pressure were considered. Genotyping of AdipoQ (G>T) gene polymorphism was done by RFLP-PCR analysis. Results The comparison of the frequency distribution of alleles and genotypes of AdipoQ (+276G>T) gene polymorphism showed a significant distribution (p<0.05) among subjects with and without CVD. The risk for CVD was high (∼9 fold) among carriers of +276T allele towards CVD predisposition. Obese women with CVD under the recessive model conferred ∼8 fold high risk (p=0.001) and +276T allele and TT genotype of non-obese women with CVD for BMI <25 also conferred ∼9 fold high risk. Hypertension also acted as a strong risk factor related to CVD (p=0.0001). Under the recessive model, hypertensive PMW with CVD conferred 7–9 fold higher risk however, normotensive women with CVD also conferred 9∼10-fold risk towards CVD predisposition. Conclusions The T allele carriers of AdipoQ gene is strongly associated with risk factors such as obesity and hypertension pertaining to cardiovascular disease. Early detection of these risk factors may serve as a CVD preventative intervention.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"47 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135967774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives Colon cancer is the most common gastrointestinal cancer worldwide with high morbidity and mortality rates. The main purpose of our study is to elucidate the interaction mechanism of the H + ion concentration effect in the CO 2 /HCO 3 − buffer system of tumor-associated Carbonic Anhydrase IX (CA IX) enzyme inhibition in the HT-29 colon cancer cell line on cell epigenetic modifications. Methods Cell culture was performed using the human colon cancer cell line HT-29. CA IX enzyme inhibitor Acetazolamide (AZA) was administered. The results of the cell viability test and inhibition were evaluated. Extracellular pH measurements were performed. Total histone protein isolation was performed and Histone H3 modifications were analyzed by ELISA method. After RNA isolation, complementary DNA (cDNA) synthesis was carried out. RT-PCR was performed to determine the gene expression levels of Hypoxia-Inducible Factor 1A (HIF1A), Enhancer of Zeste Homolog 2 (EZH2) and CA IX. Results CA IX enzyme inhibition in the HT-29 cell line decreased the expression of CA IX (p<0.05) and HIF1A (p<0.01) genes and increased the expression of the EZH2 (p<0.05). There was a significant decrease in the expression of CA IX (p<0.05) and HIF1A genes as a result of inhibition with AZA performed under hypoxic conditions. It was observed that CA IX enzyme inhibition increases the expression of the EZH2 gene by more than 3 times (p<0.01). As a result of AZA inhibition, methylation levels were observed to increase in normoxic conditions, while methylation levels were observed to decrease in hypoxic conditions. Conclusions Observing the changes in the H3 modifications and changes in the expression of CA IX, HIF1A and EZH2 genes in this study supports that CA IX enzyme inhibition plays an active role in epigenetic modifications.
摘要目的结肠癌是世界范围内最常见的胃肠道肿瘤，具有较高的发病率和死亡率。本研究的主要目的是阐明肿瘤相关碳酸酐酶IX (CA IX)酶抑制HT-29结肠癌细胞系CO 2 /HCO 3−缓冲系统中H +离子浓度效应对细胞表观遗传修饰的相互作用机制。方法采用人结肠癌细胞系HT-29进行细胞培养。给予caix酶抑制剂乙酰唑胺(AZA)。对细胞活力试验和抑制效果进行评价。进行细胞外pH测量。分离组蛋白总蛋白，ELISA法检测组蛋白H3修饰。RNA分离后，进行互补DNA合成。采用RT-PCR检测缺氧诱导因子1A (HIF1A)、Zeste同源物增强子2 (EZH2)和CA IX的基因表达水平。结果CA IX酶在HT-29细胞系中的抑制作用降低了CA IX (p<0.05)和HIF1A (p<0.01)基因的表达，增加了EZH2的表达(p<0.05)。缺氧条件下，由于AZA的抑制作用，CA IX和HIF1A基因的表达显著降低(p<0.05)。结果表明，CA IX酶抑制使EZH2基因表达量增加3倍以上(p<0.01)。由于AZA抑制，甲基化水平在常氧条件下升高，而在缺氧条件下甲基化水平降低。结论本研究通过观察H3修饰的变化以及CA IX、HIF1A和EZH2基因的表达变化，支持CA IX酶抑制在表观遗传修饰中发挥积极作用。
{"title":"Investigation of the effect of CA IX enzyme inhibition on the EZH2 gene and histone 3 modifications","authors":"İbrahim Karakus, Özen Özensoy Guler","doi":"10.1515/tjb-2023-0066","DOIUrl":"https://doi.org/10.1515/tjb-2023-0066","url":null,"abstract":"Abstract Objectives Colon cancer is the most common gastrointestinal cancer worldwide with high morbidity and mortality rates. The main purpose of our study is to elucidate the interaction mechanism of the H + ion concentration effect in the CO 2 /HCO 3 − buffer system of tumor-associated Carbonic Anhydrase IX (CA IX) enzyme inhibition in the HT-29 colon cancer cell line on cell epigenetic modifications. Methods Cell culture was performed using the human colon cancer cell line HT-29. CA IX enzyme inhibitor Acetazolamide (AZA) was administered. The results of the cell viability test and inhibition were evaluated. Extracellular pH measurements were performed. Total histone protein isolation was performed and Histone H3 modifications were analyzed by ELISA method. After RNA isolation, complementary DNA (cDNA) synthesis was carried out. RT-PCR was performed to determine the gene expression levels of Hypoxia-Inducible Factor 1A (HIF1A), Enhancer of Zeste Homolog 2 (EZH2) and CA IX. Results CA IX enzyme inhibition in the HT-29 cell line decreased the expression of CA IX (p<0.05) and HIF1A (p<0.01) genes and increased the expression of the EZH2 (p<0.05). There was a significant decrease in the expression of CA IX (p<0.05) and HIF1A genes as a result of inhibition with AZA performed under hypoxic conditions. It was observed that CA IX enzyme inhibition increases the expression of the EZH2 gene by more than 3 times (p<0.01). As a result of AZA inhibition, methylation levels were observed to increase in normoxic conditions, while methylation levels were observed to decrease in hypoxic conditions. Conclusions Observing the changes in the H3 modifications and changes in the expression of CA IX, HIF1A and EZH2 genes in this study supports that CA IX enzyme inhibition plays an active role in epigenetic modifications.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"120 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136012961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives The aim of this study is to investigate the diagnostic and prognostic characteristics of fetuin A and fetuin B in chronic hepatitis B(CHB) and HBe Ag-negative chronic infection (HCI) and the relationship between the levels of these proteins and fibrosis in CHB. Methods In this study, we examined 98 patients with CHB, 58 with HCI and 42 control groups. Fetuin A and B levels were determined via ELISA. Results Serum fetuin A and B levels were significantly higher in the control group than the hepatitis B cases (p=0.001). No significant difference in fetuin A and B levels between patients in the CHB and HCI. In the CHB, fetuin A level was significantly lower in patients with significant fibrosis than those with mild fibrosis (p=0.007). Fetuin B was lower in patients with significant fibrosis than in with mild fibrosis; however, this difference was not significant. In predicting the absence of significant fibrosis, the area under the curve was estimated as 0.855 for fetuin A and 0.866 for fetuin B using the ROC curve. Conclusions Fetuin A and B were lower in CHB and HCI compared to the control group and there was no difference between the two groups suggests that these proteins may be effective in the pathogenesis of hepatitis B-induced liver damage. Fetuin A and B, which are found to be lower in patients with significant fibrosis in CHB, can be used as non-invasive markers in the early detection of fibrosis and in the follow-up of progression to significant fibrosis.
{"title":"Fetuin A and fetuin B as an indicator of liver fibrosis in hepatitis B","authors":"Arzu Şenol, Şafak Özer Balin, Zülal Aşçı Toraman","doi":"10.1515/tjb-2022-0197","DOIUrl":"https://doi.org/10.1515/tjb-2022-0197","url":null,"abstract":"Abstract Objectives The aim of this study is to investigate the diagnostic and prognostic characteristics of fetuin A and fetuin B in chronic hepatitis B(CHB) and HBe Ag-negative chronic infection (HCI) and the relationship between the levels of these proteins and fibrosis in CHB. Methods In this study, we examined 98 patients with CHB, 58 with HCI and 42 control groups. Fetuin A and B levels were determined via ELISA. Results Serum fetuin A and B levels were significantly higher in the control group than the hepatitis B cases (p=0.001). No significant difference in fetuin A and B levels between patients in the CHB and HCI. In the CHB, fetuin A level was significantly lower in patients with significant fibrosis than those with mild fibrosis (p=0.007). Fetuin B was lower in patients with significant fibrosis than in with mild fibrosis; however, this difference was not significant. In predicting the absence of significant fibrosis, the area under the curve was estimated as 0.855 for fetuin A and 0.866 for fetuin B using the ROC curve. Conclusions Fetuin A and B were lower in CHB and HCI compared to the control group and there was no difference between the two groups suggests that these proteins may be effective in the pathogenesis of hepatitis B-induced liver damage. Fetuin A and B, which are found to be lower in patients with significant fibrosis in CHB, can be used as non-invasive markers in the early detection of fibrosis and in the follow-up of progression to significant fibrosis.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"60 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136254682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Objectives This study aimed to investigate whether a growth factor and a cytokine midkine (MK) can be a new biomarker for the diagnosis and treatment of unexplained female infertility (UFI) cases. Methods Serum (S), follicle fluid (FF), and cumulus cells (CCs) of the patients aged 20–42 years, diagnosed with male factor (MF) and UFI were used. Patients underwent Intracytoplasmic Sperm Injection (ICSI). The Anti-Müllerian Hormone (AMH) and MK levels with other hormone levels (FSH, LH, E2, PRL, INHB, TSH), the oocyte and embryo qualities, the fertilization and pregnancy rates, and cumulus cells (Cell number and ultrastructure, apoptosis rate) were evaluated. Student-T-test was performed and p<0.05 was considered statistically significant. Results and Discussion The lowest numbers of CCs were found at UFI (p<0.05). The lowest apoptosis rate with the highest CC viability rate was evaluated at MF (p<0.05). The lowest AMH and MK levels (S, FF) were detected at UFI in comparison to MF (p<0.05). MK and AMH levels of non-pregnant subjects were much lower than pregnant subjects (p<0.05). In addition, these levels were lower in the subjects above 35 age (p<0.05). Structural analysis of CCs showed that the number of lytic cells with cell remnants and apoptotic bodies was higher in non-pregnant subjects. It seems that MK did not show any resistance to both AMH and apoptosis. Conclusions MK can not be accepted as a new biomarker for the diagnosis and treatment monitoring of UFI cases.
{"title":"Midkine can not be accepted as a new biomarker for unexplained female infertility","authors":"Mine Ergüven, Semra Kahraman, Caroline Pirkevi, Tülay İrez","doi":"10.1515/tjb-2023-0055","DOIUrl":"https://doi.org/10.1515/tjb-2023-0055","url":null,"abstract":"Abstract Objectives This study aimed to investigate whether a growth factor and a cytokine midkine (MK) can be a new biomarker for the diagnosis and treatment of unexplained female infertility (UFI) cases. Methods Serum (S), follicle fluid (FF), and cumulus cells (CCs) of the patients aged 20–42 years, diagnosed with male factor (MF) and UFI were used. Patients underwent Intracytoplasmic Sperm Injection (ICSI). The Anti-Müllerian Hormone (AMH) and MK levels with other hormone levels (FSH, LH, E2, PRL, INHB, TSH), the oocyte and embryo qualities, the fertilization and pregnancy rates, and cumulus cells (Cell number and ultrastructure, apoptosis rate) were evaluated. Student-T-test was performed and p<0.05 was considered statistically significant. Results and Discussion The lowest numbers of CCs were found at UFI (p<0.05). The lowest apoptosis rate with the highest CC viability rate was evaluated at MF (p<0.05). The lowest AMH and MK levels (S, FF) were detected at UFI in comparison to MF (p<0.05). MK and AMH levels of non-pregnant subjects were much lower than pregnant subjects (p<0.05). In addition, these levels were lower in the subjects above 35 age (p<0.05). Structural analysis of CCs showed that the number of lytic cells with cell remnants and apoptotic bodies was higher in non-pregnant subjects. It seems that MK did not show any resistance to both AMH and apoptosis. Conclusions MK can not be accepted as a new biomarker for the diagnosis and treatment monitoring of UFI cases.","PeriodicalId":92463,"journal":{"name":"Turk biyokimya dergisi = Turkish journal of biochemistry","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135303887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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