Isabel Cristina Teixeira Proença, Tamires Marques de Abreu, Jessica Pereira Marinho, Malena Rostirola Miri, Gustavo Fernandes Vasques, Luiz Fernando Lopes, Manuela Dos Santos, Vanessa Duarte Ortiz, Patrick Türck, Alex Sander da Rosa Araujo, Claudia Funchal, Daniela Pochmann, Caroline Dani
This study aimed to evaluate the effects of purple grape juice consumption in pregnancy on oxidative stress parameters in Wistar rat fetuses. Twenty-four pregnant rats were divided into five groups: control group, indomethacin group (received a single dose of indomethacin in DG20), group grape juice DG14 (received an amount for 14 days/first and second gestational trim), group grape juice DG20 (received a dose throughout the gestational period), group grape juice two doses (received two doses, at morning and afternoon). On the 20th day of pregnancy (DG20), rats were anesthetized, and a cesarean section was performed to obtain the fetuses. A sample of liver, heart, and total brain of fetuses was collected for oxidative stress analyses. Values P<0.05 were considered significant. In fetuses' heart, we observed that the grape juice two dose group decreased sulfhydryl and increased SOD. In the liver, the grape juice decreased TBARS and SOD. There was a decrease in carbonyl and sulfhydryl in the indomethacin and grape juice one dose groups in the brain. We conclude that indomethacin altered oxidative stress parameters only in the fetal brain, and grape juice was presented as an important modulator of antioxidant capacity when consumed in a dose.
{"title":"The effect of consumption of purple grape juice in the gestational period on oxidative stress parameters in <i>Wistar</i> rat foetuses.","authors":"Isabel Cristina Teixeira Proença, Tamires Marques de Abreu, Jessica Pereira Marinho, Malena Rostirola Miri, Gustavo Fernandes Vasques, Luiz Fernando Lopes, Manuela Dos Santos, Vanessa Duarte Ortiz, Patrick Türck, Alex Sander da Rosa Araujo, Claudia Funchal, Daniela Pochmann, Caroline Dani","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>This study aimed to evaluate the effects of purple grape juice consumption in pregnancy on oxidative stress parameters in <i>Wistar</i> rat fetuses. Twenty-four pregnant rats were divided into five groups: control group, indomethacin group (received a single dose of indomethacin in DG20), group grape juice DG14 (received an amount for 14 days/first and second gestational trim), group grape juice DG20 (received a dose throughout the gestational period), group grape juice two doses (received two doses, at morning and afternoon). On the 20th day of pregnancy (DG20), rats were anesthetized, and a cesarean section was performed to obtain the fetuses. A sample of liver, heart, and total brain of fetuses was collected for oxidative stress analyses. Values P<0.05 were considered significant. In fetuses' heart, we observed that the grape juice two dose group decreased sulfhydryl and increased SOD. In the liver, the grape juice decreased TBARS and SOD. There was a decrease in carbonyl and sulfhydryl in the indomethacin and grape juice one dose groups in the brain. We conclude that indomethacin altered oxidative stress parameters only in the fetal brain, and grape juice was presented as an important modulator of antioxidant capacity when consumed in a dose.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 3","pages":"60-68"},"PeriodicalIF":0.0,"publicationDate":"2021-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784795/pdf/ijbmb0012-0060.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39876960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Indoleamine 2,3-dioxygenase is an important enzyme in human, which plays roles in the rate-limiting first step in tryptophan catabolism. Indoleamine 2,3-dioxygenase is currently studied by research groups worldwide. Association between Indoleamine 2,3-dioxygenase and medial disorders in clinical medicine is proposed and is an interesting topic in clinical biochemical reasearch. Here, the author briefly reviews and discusses on the available important scientific reports on Indoleamine 2,3-dioxygenase from tropical Southeast Asia.
{"title":"Clinical biomedical research of indoleamine 2,3-dioxygenase: update on current available reports from Southeast Asia.","authors":"Beuy Joob, Sora Yasri, Viroj Wiwanitkit","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Indoleamine 2,3-dioxygenase is an important enzyme in human, which plays roles in the rate-limiting first step in tryptophan catabolism. Indoleamine 2,3-dioxygenase is currently studied by research groups worldwide. Association between Indoleamine 2,3-dioxygenase and medial disorders in clinical medicine is proposed and is an interesting topic in clinical biochemical reasearch. Here, the author briefly reviews and discusses on the available important scientific reports on Indoleamine 2,3-dioxygenase from tropical Southeast Asia.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 3","pages":"55-59"},"PeriodicalIF":0.0,"publicationDate":"2021-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784796/pdf/ijbmb0012-0055.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39876959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective: Observing the pathogens and drug-resistance within hospital-acquired pneumonia (HAP) in an emergency intensive care unit (EICU) to provide a reference for clinically reasonable use of antibiotics.
Methods: Sixty-two patients with HAP in Tianjin Medical University General Hospital from January 2017 to May 2019 were analyzed retrospectively. Bacterial identification and susceptibility were reviewed.
Results: One hundred and thirty-seven strains of pathogenic bacteria were isolated from 62 patients, with 97.1% Gram-negative and only 2.9% Gram-positive. There were also six fungal isolates. The most common pathogens were Acinetobacter baumannii, accounting for 30.8% of all isolates, followed by Klebisella spp, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Escherichia coli. Acinetobacter baumannii was poorly susceptible to piperacillin-tazobactam, cefepime, Amoxicillin+clavulonic acid, ciprofloxacin. However, the isolates were sensitive to Tigecycline, so as the isolates of Klebisella spp. Pseudomonas aeruginosa was mostly sensitive to Amikacin, followed by Tobramycin. All of the isolates of Staphylococcus aureus were susceptible to Linezolid, Tigecycline and Vancomycin.
Conclusions: Gram-negative bacteria especially Acinetobacter baumannii, are the main pathogens for HAP in the observed EICU. The variety of pathogens should be monitored at regular intervals to improve resistance issues and therapeutic effect.
{"title":"Pathogens and drug-resistance of hospital-acquired pneumonia in an EICU in Tianjin, China.","authors":"Yanan Zhang, Songtao Shou","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>Observing the pathogens and drug-resistance within hospital-acquired pneumonia (HAP) in an emergency intensive care unit (EICU) to provide a reference for clinically reasonable use of antibiotics.</p><p><strong>Methods: </strong>Sixty-two patients with HAP in Tianjin Medical University General Hospital from January 2017 to May 2019 were analyzed retrospectively. Bacterial identification and susceptibility were reviewed.</p><p><strong>Results: </strong>One hundred and thirty-seven strains of pathogenic bacteria were isolated from 62 patients, with 97.1% Gram-negative and only 2.9% Gram-positive. There were also six fungal isolates. The most common pathogens were Acinetobacter baumannii, accounting for 30.8% of all isolates, followed by Klebisella spp, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Escherichia coli. Acinetobacter baumannii was poorly susceptible to piperacillin-tazobactam, cefepime, Amoxicillin+clavulonic acid, ciprofloxacin. However, the isolates were sensitive to Tigecycline, so as the isolates of Klebisella spp. Pseudomonas aeruginosa was mostly sensitive to Amikacin, followed by Tobramycin. All of the isolates of Staphylococcus aureus were susceptible to Linezolid, Tigecycline and Vancomycin.</p><p><strong>Conclusions: </strong>Gram-negative bacteria especially Acinetobacter baumannii, are the main pathogens for HAP in the observed EICU. The variety of pathogens should be monitored at regular intervals to improve resistance issues and therapeutic effect.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 2","pages":"49-54"},"PeriodicalIF":0.0,"publicationDate":"2021-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8166653/pdf/ijbmb0012-0049.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39058127","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shabhat Rasool, Rubiya Dar, Mosin S Khan, Sheikh Gazalla Ayoub, Sabia Rashid, Muneeb U Rehman, Tariq Jan, Meenu A Qureshi, Khurshid I Andrabi
High Myopia (HM) is a common complex-trait eye disorder. There is essential evidence that genetic factors play a significant role in the development of nonsyndromic high myopia. Identification of susceptibility genes of high myopia will shed light on the pathophysiological mechanism underlying their genesis. This was a case control study examining the prospect of association of DLGAP1, EMILIN2 & MYOM1 genes on MYP2 locus in purely ethnic (Kashmiri) population representing a homogeneous cohort. Genomic DNA was extracted using phenol chloroform and salting out method. Extracted DNA was genotyped for polymorphic variations in MYOM1, EMILIN2 and DLGAP1 genes involving Sanger di-deoxy method. Allele frequencies were tested for Hardy-Weinberg disequilibrium in 224 cases and compared with 220 emmetropic controls. In DLGAP1, documented single nucleotide polymorphism (SNP); Pro517Pro was observed. A previously reported Asn451Asn SNP was observed in EMILIN2. MYOM1 showed five polymorphic variations; two in coding region (Gly333Gly & Gly341Ala) and three intronic (c.1022+23, G>A; c.3418+44 G>T & c.3418+65; C>G). All of the elucidated SNPs were having statistical significant role in increasing or decreasing the risk of disease. Although not statistically significant, a novel Glu507Lys SNP was observed in DLGAP1 (P>0.05). In silico predictions showed MYOM1 Gly341Ala to be benign & tolerated substitution while as DLGAP1 Glu507Lys to be possibly damaging substitution. The studied SNPs followed Over-Dominant, Recessive and Co-Dominant mode of inheritance with specific haplotypes associated with the disease. Our study reveals the involvement of MYP2 locus candidate gene polymorphism in the pathogenesis of HM.
{"title":"MYP2 locus genes: Sequence variations, genetic association studies and haplotypic association in patients with High Myopia.","authors":"Shabhat Rasool, Rubiya Dar, Mosin S Khan, Sheikh Gazalla Ayoub, Sabia Rashid, Muneeb U Rehman, Tariq Jan, Meenu A Qureshi, Khurshid I Andrabi","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>High Myopia (HM) is a common complex-trait eye disorder. There is essential evidence that genetic factors play a significant role in the development of nonsyndromic high myopia. Identification of susceptibility genes of high myopia will shed light on the pathophysiological mechanism underlying their genesis. This was a case control study examining the prospect of association of <i>DLGAP1, EMILIN2 & MYOM1</i> genes on <i>MYP2</i> locus in purely ethnic (Kashmiri) population representing a homogeneous cohort. Genomic DNA was extracted using phenol chloroform and salting out method. Extracted DNA was genotyped for polymorphic variations in <i>MYOM1, EMILIN2 and DLGAP1</i> genes involving Sanger di-deoxy method. Allele frequencies were tested for Hardy-Weinberg disequilibrium in 224 cases and compared with 220 emmetropic controls. In <i>DLGAP1</i>, documented single nucleotide polymorphism (SNP); <i>Pro517Pro</i> was observed. A previously reported <i>Asn451Asn</i> SNP was observed in <i>EMILIN2</i>. <i>MYOM1</i> showed five polymorphic variations; two in coding region (<i>Gly333Gly & Gly341Ala</i>) and three intronic (c.1022+23, G>A; c.3418+44 G>T & c.3418+65; C>G). All of the elucidated SNPs were having statistical significant role in increasing or decreasing the risk of disease. Although not statistically significant, a novel <i>Glu507Lys</i> SNP was observed in <i>DLGAP1</i> (P>0.05). In silico predictions showed <i>MYOM1 Gly341Ala</i> to be benign & tolerated substitution while as <i>DLGAP1 Glu507Lys</i> to be possibly damaging substitution. The studied SNPs followed Over-Dominant, Recessive and Co-Dominant mode of inheritance with specific haplotypes associated with the disease. Our study reveals the involvement of <i>MYP2</i> locus candidate gene polymorphism in the pathogenesis of HM.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 1","pages":"35-48"},"PeriodicalIF":0.0,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8012819/pdf/ijbmb0012-0035.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25567592","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abraham A Paul, Omolaja R Osoniyi, Queensley A Eghianruwa, Olayemi G Oparinde, Adetunji S Adesina
The multifunctional nature of antioxidant peptides makes them more attractive candidates as dietary ingredients in health maintenance. Therefore, food protein-derived antioxidant peptides are continuously investigated. This study investigated the in vitro antioxidant properties of hydrolysate and ultrafiltered peptide fractions of Pachymelania aurita and Tympanatonus fuscatus var radula-two commonly consumed marine molluscs known as periwinkles in southern Nigeria. Simulated gastrointestinal digestion (SGID) of soluble proteins of T. fuscatus and P. aurita was carried out using pepsin, trypsin and chymotrypsin, and the SGID hydrolysates were fractionated using a 3 kDa membrane filter. The hydrolysates and their fractions were investigated for anti-lipid peroxidation, hydroxyl radical scavenging activity (HRSA), ferric reducing antioxidant property (FRAP) and metal chelation activity, and they demonstrated clear antioxidant properties in all the assay models used. Low molecular weight fractions of the hydrolysates demonstrated more potent antioxidant activity than higher molecular weight fractions. This is profound in the metal chelation assay, where low molecular weight peptide fractions, T ≤ 3 kDa and P ≤ 3 kDa (IC50 values of 8.10 ± 0.011 and 5.56 ± 0.50 µg/ml respectively) had activity that is similar to that of EDTA (11.84 ± 0.89 µg/ml). Similar activity effects were observed in other assays where there was about 3-fold higher activity in low molecular weight fractions. These results demonstrate the presence of antioxidant peptide(s) in the protein hydrolysates of the periwinkles.
{"title":"Antioxidant activity of the simulated gastro-intestinal digestion hydrolysate of two edible Nigerian marine molluscs: <i>Tympanatonus fuscatus</i> var radula (L.) and <i>Pachymelania aurita</i> (M.).","authors":"Abraham A Paul, Omolaja R Osoniyi, Queensley A Eghianruwa, Olayemi G Oparinde, Adetunji S Adesina","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The multifunctional nature of antioxidant peptides makes them more attractive candidates as dietary ingredients in health maintenance. Therefore, food protein-derived antioxidant peptides are continuously investigated. This study investigated the <i>in vitro</i> antioxidant properties of hydrolysate and ultrafiltered peptide fractions of <i>Pachymelania aurita</i> and <i>Tympanatonus fuscatus</i> var radula-two commonly consumed marine molluscs known as periwinkles in southern Nigeria. Simulated gastrointestinal digestion (SGID) of soluble proteins of <i>T. fuscatus</i> and <i>P. aurita</i> was carried out using pepsin, trypsin and chymotrypsin, and the SGID hydrolysates were fractionated using a 3 kDa membrane filter. The hydrolysates and their fractions were investigated for anti-lipid peroxidation, hydroxyl radical scavenging activity (HRSA), ferric reducing antioxidant property (FRAP) and metal chelation activity, and they demonstrated clear antioxidant properties in all the assay models used. Low molecular weight fractions of the hydrolysates demonstrated more potent antioxidant activity than higher molecular weight fractions. This is profound in the metal chelation assay, where low molecular weight peptide fractions, T ≤ 3 kDa and P ≤ 3 kDa (IC<sub>50</sub> values of 8.10 ± 0.011 and 5.56 ± 0.50 µg/ml respectively) had activity that is similar to that of EDTA (11.84 ± 0.89 µg/ml). Similar activity effects were observed in other assays where there was about 3-fold higher activity in low molecular weight fractions. These results demonstrate the presence of antioxidant peptide(s) in the protein hydrolysates of the periwinkles.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 1","pages":"8-16"},"PeriodicalIF":0.0,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8012820/pdf/ijbmb0012-0008.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25567590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alkaline phosphatase is an enzyme that converts para-nitrophenyl phosphate to para-nitrophenol (yellow coloured) in 2-amino, 2-methyl, 1-propanol buffer at pH 10.5. However, when this protocol is applied to the in vitro cellular model systems to estimate alkaline phosphatase activity, it tends to generate clumps of genomic DNA, leading to inaccurate pipetting for protein estimation. The aim of the study was to introduce minor modifications in the existing protocol to make it simple, cost-effective, with minimal labor-intensive procedures while estimating alkaline phosphatase activity in cellular model systems. The genomic DNA clumps were dissolved by depurination (adding 0.2 N HCl) and fragmentation (adding 0.2 N NaOH) during enzyme estimation. Moreover, these minor modifications have been standardized and optimized extensively by using serum samples (rich source of alkaline phosphatase), hFOB/ER9 (human Fetal osteoblastic cell) and HepG2 cells. Our results suggest that the modification incorporated in previously published method was robust enough to estimate ALP activity and protein concentration accurately. There was no significant variation in ALP activity estimated after modification (P > 0.05). This innovative approach could be beneficial for a researcher by providing an easy, cost effective and less labor-intensive solution for estimation of enzymatic activity in cellular model systems.
碱性磷酸酶是一种在pH为10.5的2-氨基,2-甲基,1-丙醇缓冲液中将对硝基苯基磷酸盐转化为对硝基苯酚(黄色)的酶。然而,当该方案应用于体外细胞模型系统来估计碱性磷酸酶活性时,它往往会产生基因组DNA团块,导致不准确的移液来估计蛋白质。该研究的目的是在现有的方案中引入微小的修改,使其简单,具有成本效益,在估计细胞模型系统中的碱性磷酸酶活性时,使用最少的劳动密集型程序。基因组DNA团块在酶测定时通过去嘌呤(加入0.2 N HCl)和片段化(加入0.2 N NaOH)溶解。此外,这些微小的修饰已经通过血清样本(碱性磷酸酶的丰富来源)、hFOB/ER9(人胎儿成骨细胞)和HepG2细胞进行了广泛的标准化和优化。我们的结果表明,先前发表的方法中包含的修饰足以准确估计ALP活性和蛋白质浓度。改良后ALP活性无显著变化(P > 0.05)。这种创新的方法可以为研究人员提供一种简单,成本效益高,劳动强度低的解决方案,用于估计细胞模型系统中的酶活性。
{"title":"An innovative and cost-effective way to estimate alkaline phosphatase activity in <i>in vitro</i> cellular model systems.","authors":"Poonam Kanta, Tulikalipi Ghosh, Anit Kaur, Thungapathra Muthukumarappa","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Alkaline phosphatase is an enzyme that converts para-nitrophenyl phosphate to para-nitrophenol (yellow coloured) in 2-amino, 2-methyl, 1-propanol buffer at pH 10.5. However, when this protocol is applied to the in vitro cellular model systems to estimate alkaline phosphatase activity, it tends to generate clumps of genomic DNA, leading to inaccurate pipetting for protein estimation. The aim of the study was to introduce minor modifications in the existing protocol to make it simple, cost-effective, with minimal labor-intensive procedures while estimating alkaline phosphatase activity in cellular model systems. The genomic DNA clumps were dissolved by depurination (adding 0.2 N HCl) and fragmentation (adding 0.2 N NaOH) during enzyme estimation. Moreover, these minor modifications have been standardized and optimized extensively by using serum samples (rich source of alkaline phosphatase), hFOB/ER9 (human Fetal osteoblastic cell) and HepG2 cells. Our results suggest that the modification incorporated in previously published method was robust enough to estimate ALP activity and protein concentration accurately. There was no significant variation in ALP activity estimated after modification (P > 0.05). This innovative approach could be beneficial for a researcher by providing an easy, cost effective and less labor-intensive solution for estimation of enzymatic activity in cellular model systems.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 1","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8012822/pdf/ijbmb0012-0001.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25567589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Syamalima Dube, Lynn Abbott, Samender Randhawa, Yingli Fan, Joseph W Sanger, Jean M Sanger, Bernard J Poiesz, Dipak K Dube
In mammals, there are four tropomyosin (TPM) genes (TPM1, TPM2, TPM3, and TPM4) each of which generate a multitude of alternatively spliced mRNAs. TPM isoform diversity in bovine unlike in humans are not well characterized. The purpose of this investigation is to perform an extensive analysis of the expression of both transcripts and corresponding protein of sarcomeric TPMs in bovine strated muscles. We have cloned and sequenced the transcripts of the sarcomeric isoform of the TPM4 gene designated as TPM4α as well as a new splice variant TPM4ε from bovine striated muscles. Additionally, we have determined the expression of various sarcomeric TPM isoforms and TPM4ε in bovine heart and skeletal muscles. Relative expression as well as absolute copy number determination by qRT-PCR suggests that TPM1α expression is significantly higher in bovine cardiac muscle, whereas TPM2α is higher in skeletal muscle. The relative expression of TPM3α in bovine heart and skeletal muscle is very similar. The relative expression of TPM4α and TPM4ε is higher in bovine heart and skeletal muscle, respectively. We have evaluated the protein expression levels of various TPM isoforms by 2D western blot analyses in commercially available protein extracts of heart and skeletal muscles with the CH1 monoclonal antibody against TPM. Protein from each CH1-positive spot was extracted for LC-MS/MS analyses, which show that bovine heart extract contains 91.66% TPM1 and 8.33% TPM2, whereas skeletal muscle extract contains 57% TPM1 and 42.87% TPM2. We have failed to detect the presence of unique peptide(s) for TPM3α, TPM4α, and TPM4ε.
在哺乳动物中,有四种原肌球蛋白(TPM)基因(TPM1, TPM2, TPM3和TPM4),每个基因都产生大量的选择性剪接mrna。与人类不同,牛的TPM异构体多样性并没有很好地表征。本研究的目的是对牛层状肌中肌性TPMs的转录本和相应蛋白的表达进行广泛的分析。我们克隆并测序了牛横纹肌中TPM4基因的肌肉异构体TPM4α和一个新的剪接变体TPM4ε的转录本。此外,我们还确定了牛心脏和骨骼肌中各种肌肉异构体TPM和TPM4ε的表达。qRT-PCR检测的相对表达量和绝对拷贝数表明,TPM1α在牛心肌中的表达量显著高于TPM2α在骨骼肌中的表达量。TPM3α在牛心脏和骨骼肌中的相对表达量非常相似。TPM4α和TPM4ε在牛心脏和骨骼肌中的相对表达量较高。我们用抗TPM的CH1单克隆抗体对市售的心脏和骨骼肌蛋白提取物进行了2D western blot分析,评估了各种TPM亚型的蛋白表达水平。提取每个ch1阳性点的蛋白进行LC-MS/MS分析,结果表明,牛心脏提取物TPM1含量为91.66%,TPM2含量为8.33%,骨骼肌提取物TPM1含量为57%,TPM2含量为42.87%。我们没有检测到TPM3α、TPM4α和TPM4ε的独特肽的存在。
{"title":"Identification of a novel TPM4 isoform transcript and comparison to the expression of other tropomyosin isoforms in bovine cardiac and skeletal muscles.","authors":"Syamalima Dube, Lynn Abbott, Samender Randhawa, Yingli Fan, Joseph W Sanger, Jean M Sanger, Bernard J Poiesz, Dipak K Dube","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>In mammals, there are four tropomyosin (TPM) genes (TPM1, TPM2, TPM3, and TPM4) each of which generate a multitude of alternatively spliced mRNAs. TPM isoform diversity in bovine unlike in humans are not well characterized. The purpose of this investigation is to perform an extensive analysis of the expression of both transcripts and corresponding protein of sarcomeric TPMs in bovine strated muscles. We have cloned and sequenced the transcripts of the sarcomeric isoform of the TPM4 gene designated as TPM4α as well as a new splice variant TPM4ε from bovine striated muscles. Additionally, we have determined the expression of various sarcomeric TPM isoforms and TPM4ε in bovine heart and skeletal muscles. Relative expression as well as absolute copy number determination by qRT-PCR suggests that TPM1α expression is significantly higher in bovine cardiac muscle, whereas TPM2α is higher in skeletal muscle. The relative expression of TPM3α in bovine heart and skeletal muscle is very similar. The relative expression of TPM4α and TPM4ε is higher in bovine heart and skeletal muscle, respectively. We have evaluated the protein expression levels of various TPM isoforms by 2D western blot analyses in commercially available protein extracts of heart and skeletal muscles with the CH1 monoclonal antibody against TPM. Protein from each CH1-positive spot was extracted for LC-MS/MS analyses, which show that bovine heart extract contains 91.66% TPM1 and 8.33% TPM2, whereas skeletal muscle extract contains 57% TPM1 and 42.87% TPM2. We have failed to detect the presence of unique peptide(s) for TPM3α, TPM4α, and TPM4ε.</p>","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"12 1","pages":"17-34"},"PeriodicalIF":0.0,"publicationDate":"2021-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8012821/pdf/ijbmb0012-0017.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25567591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.33545/26646501.2021.v3.i1a.18
A. Chatterjee
{"title":"Neutrophil lymphocyte ratio (NLR) is inversely proportional to cycle threshold value of orf1a gene in RTPCR may be used as biomarker in Covid19 patients","authors":"A. Chatterjee","doi":"10.33545/26646501.2021.v3.i1a.18","DOIUrl":"https://doi.org/10.33545/26646501.2021.v3.i1a.18","url":null,"abstract":"","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80079819","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}
Pub Date : 2021-01-01DOI: 10.33545/26646501.2021.v3.i1a.22
V. Phuong, Vo Van Vu, Nguyen Hoai Thanh, Lê Thị Thùy Dung, Vo Thi Nhi, Äo Thi Ngoc Trinh
{"title":"Density and distribution of calcium oxalate crystals in vegetative organs of “green” wild taro (Colocasia esculenta (L.) Schott.)","authors":"V. Phuong, Vo Van Vu, Nguyen Hoai Thanh, Lê Thị Thùy Dung, Vo Thi Nhi, Äo Thi Ngoc Trinh","doi":"10.33545/26646501.2021.v3.i1a.22","DOIUrl":"https://doi.org/10.33545/26646501.2021.v3.i1a.22","url":null,"abstract":"","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75008035","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}
Pub Date : 2021-01-01DOI: 10.33545/26646501.2021.v3.i1a.19
S. Menon
{"title":"Bioinformatics approaches for cross-species liver cancer analysis based on single-cell RNA sequencing data in rats mice and human insights causative genes and pathways","authors":"S. Menon","doi":"10.33545/26646501.2021.v3.i1a.19","DOIUrl":"https://doi.org/10.33545/26646501.2021.v3.i1a.19","url":null,"abstract":"","PeriodicalId":13891,"journal":{"name":"International journal of biochemistry and molecular biology","volume":"27 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83575979","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}