Pub Date : 2020-06-01DOI: 10.4236/ajmb.2020.103015
O. Goselle, G. Y. Ajiji, A. Jambol, J. Sunday, Ojochemi Sunday Idoko, S. S. Udoh, Oliseemeka Charles Ejete, Y. M. Ahmadu, H. Awobode, G. Imandeh, B. Matur
The discovery of Plasmodium parasites and its incrimination as the principal cause of malaria in humans has continued to excite researchers towards inventing possible easier methods of diagnosing and identifying these pathological agents in order to mitigate, control and eliminate its continuous scourge to humanity. Currently, three diagnostic methods have been proposed, but agreements as to whether the level of parasitaemia in an individual could connote likely confirmations in the three methods i.e. gold standard, RDTs’ and PCR/NESTED PCR, have continued to be a subject of debate. To lay to rest the debate as reported in many studies, we collected blood samples from 100 symptomatic patients who reported to the Jos-Nigeria hospital and using the gold standard methods, we were able to confirm that 30 (30%) samples out of the 100 blood samples collected were positive to P. falciparum, chiefly recorded among duffy-negative Africans. Excited with our findings, we prepared the thick blood films for each sample and used it to estimate the levels of parasitaemia (parasites density) per μl of blood (i.e. 1+; 2+; 3+ and 4+) per 100 high power fields (|HPF). We then subjected the individually confirmed parasite density samples to the other two methods i.e. Rapid Diagnostic Test (one-step RTD and optimal-IT® RDT) and to molecular assay (PCR and the nested PCR). Interestingly, of the 30 positive samples, 18 (60%) were confirmed positive to the one-step and optimal-IT® RDTS, while 3 (30%) out of the 10 (100%) samples of various parasite density subjected to molecular assay (PCR and the nested PCR) were positive to only P. falciparum. Statistical analysis of variance based on single factor computed using SPSS indicates a no significant difference (P > 0.05) in the parasitaemia levels of the four groups/categories of patients; i.e. variance ratio of 0.011976 calculated was less than F-critical (2.816466) at 5% (0.05). Whereas gold standard could be considered as the optimal method, for the PCR/NESTED PCR, the sensitivity is dependent on high level of parasitaemia.
{"title":"Could the Level in Parasitaemia of Plasmodium Determine Sensitivity to Various Diagnostic Tests?","authors":"O. Goselle, G. Y. Ajiji, A. Jambol, J. Sunday, Ojochemi Sunday Idoko, S. S. Udoh, Oliseemeka Charles Ejete, Y. M. Ahmadu, H. Awobode, G. Imandeh, B. Matur","doi":"10.4236/ajmb.2020.103015","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103015","url":null,"abstract":"The discovery of Plasmodium parasites and its incrimination as the principal cause of malaria in humans has continued to excite researchers towards inventing possible easier methods of diagnosing and identifying these pathological agents in order to mitigate, control and eliminate its continuous scourge to humanity. Currently, three diagnostic methods have been proposed, but agreements as to whether the level of parasitaemia in an individual could connote likely confirmations in the three methods i.e. gold standard, RDTs’ and PCR/NESTED PCR, have continued to be a subject of debate. To lay to rest the debate as reported in many studies, we collected blood samples from 100 symptomatic patients who reported to the Jos-Nigeria hospital and using the gold standard methods, we were able to confirm that 30 (30%) samples out of the 100 blood samples collected were positive to P. falciparum, chiefly recorded among duffy-negative Africans. Excited with our findings, we prepared the thick blood films for each sample and used it to estimate the levels of parasitaemia (parasites density) per μl of blood (i.e. 1+; 2+; 3+ and 4+) per 100 high power fields (|HPF). We then subjected the individually confirmed parasite density samples to the other two methods i.e. Rapid Diagnostic Test (one-step RTD and optimal-IT® RDT) and to molecular assay (PCR and the nested PCR). Interestingly, of the 30 positive samples, 18 (60%) were confirmed positive to the one-step and optimal-IT® RDTS, while 3 (30%) out of the 10 (100%) samples of various parasite density subjected to molecular assay (PCR and the nested PCR) were positive to only P. falciparum. Statistical analysis of variance based on single factor computed using SPSS indicates a no significant difference (P > 0.05) in the parasitaemia levels of the four groups/categories of patients; i.e. variance ratio of 0.011976 calculated was less than F-critical (2.816466) at 5% (0.05). Whereas gold standard could be considered as the optimal method, for the PCR/NESTED PCR, the sensitivity is dependent on high level of parasitaemia.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49494722","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 : 2020-06-01DOI: 10.4236/ajmb.2020.103012
Farag A. Bleiblo, P. Michael, C. Ramana, T. Tai, J. Parrillo, Anand Kumar, Aseem Kumar
Although much progress has been made in identifying the signaling pathways that mediate viral RNA-induced apoptosis and activation of interferon-stimulated genes, the role that bacterial RNA plays in regulating these responses has remained undetermined. Herein, we identified bacterial RNA as a novel inducer of the apoptotic cell death. Unlike the parental cells, STAT1 and STAT2 mutants display apoptotic defects which were reversed by restoring the expression of wild type proteins. While STAT1 mutants lacking tyrosine-701 or a functional SH2 domain were effective as the wild-type protein in restoring the apoptotic response, the mutant carrying a point mutation at serine-727 of STAT1 was resistant to bacterial RNA-induced apoptosis. We also determined that the lack of apoptosis in the STAT1 and STAT2 mutants was correlated with the constitutive and inducible activation of apoptosis regulating proteins. Furthermore, we show that bacterial RNA induces transcriptional activation of STAT1, STAT2, IRF1, and ISGF3, which was impaired in STAT1 or STAT2 mutants. These observations suggested that the participation of STATs in regulating the apoptotic response is independent of their downstream functions as cytokine-induced transcriptional activators. In addition to bacterial immunity, the results presented here may also have implications in cellular pathophysiology and RNA-based therapy.
{"title":"STAT1 and STAT2 Null Cells Are Resistant to RNA-Induced Apoptosis Due to Deficiency in Constitutive and Inducible Apoptosis-Regulating Genes","authors":"Farag A. Bleiblo, P. Michael, C. Ramana, T. Tai, J. Parrillo, Anand Kumar, Aseem Kumar","doi":"10.4236/ajmb.2020.103012","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103012","url":null,"abstract":"Although much progress has been made in identifying the signaling pathways that mediate viral RNA-induced apoptosis and activation of interferon-stimulated genes, the role that bacterial RNA plays in regulating these responses has remained undetermined. Herein, we identified bacterial RNA as a novel inducer of the apoptotic cell death. Unlike the parental cells, STAT1 and STAT2 mutants display apoptotic defects which were reversed by restoring the expression of wild type proteins. While STAT1 mutants lacking tyrosine-701 or a functional SH2 domain were effective as the wild-type protein in restoring the apoptotic response, the mutant carrying a point mutation at serine-727 of STAT1 was resistant to bacterial RNA-induced apoptosis. We also determined that the lack of apoptosis in the STAT1 and STAT2 mutants was correlated with the constitutive and inducible activation of apoptosis regulating proteins. Furthermore, we show that bacterial RNA induces transcriptional activation of STAT1, STAT2, IRF1, and ISGF3, which was impaired in STAT1 or STAT2 mutants. These observations suggested that the participation of STATs in regulating the apoptotic response is independent of their downstream functions as cytokine-induced transcriptional activators. In addition to bacterial immunity, the results presented here may also have implications in cellular pathophysiology and RNA-based therapy.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47395874","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 : 2020-06-01DOI: 10.4236/ajmb.2020.103014
B. Okpa, G. Gberikon, C. Aguoru, I. O. Ogbonna
Studies on ESBL-producing and multi-drug resistance of Salmonella serovars distributed in Benue State were investigated. A total of four hundred and twenty (420) clinical stool samples, seventy (70) from each local government area were randomly collected from selected hospitals and analyzed for the presence of Salmonella spp. The isolates were characterized using Gram staining and biochemical tests. The result of AP120E biochemical test strip which contained dehydrated bacterial media and biochemical reagents in twenty (20) separate compartments. The result was obtained by evaluation of the compartments due to observed changes in after 24 hours where others were read by adding up reagents (Ferric chloride, Kovacs V.P reagents). The results were analyzed afterwards in accordance with the manufacturer’s software and positive results with ≥89% potential were confirmed as Salmonella spp. Amplified plasmids derived from 18 Salmonella strains recognized were made up of 23,130 base pairs. ESBL (Extended-spectrum beta-lactamases) genes were located on the plasmids. Two of the ESBL genes found were TEM genes and CTX-M (415 bp). Strains such as S. enterica Typhimurium-CP014981.1, S. enterica Enteritidis-CP007325.2, S. enterica Typhimurium-CP024619.1, S. enterica Typhimurium-CP023166.1, S. bongori-FR877557 and S. enterica Enteritidis-TY1 possessed TEM genes where as S. enterica Heidelberg-CP019176.1, S. enterica Typhi-AL513382.1 and S. enterica Typhimurium-MH196335.1 possessed CTX-M. Antibiotic resistance testing was performed using Kirby-Bauer disc diffusion method. The overall percentage susceptibility of the eight antibiotics tested on Salmonella serovars isolates shows that GEN had the highest % susceptibility of 100% followed by NIT (72.2%) and COT (66.7%) before and after plasmid curing. % susceptibility was lower before curing than after curing in CXC, CHL and TET. It was low (5.6%) in ERY while AUG recorded 0% susceptibility. Differences observed in curing status were insignificant (T = 0.33, P > 0.05). The presence of ESBL-producing and multi-drug resistant Salmonella serovars indicates an infection which presents a foremost peril to public health since such infections may be intricate to take care of and may consequently result in death of the infected patients. Constant periodic examination and prevention of drug abuse of antibiotics will assist in ensuring that this trend is curtailed especially in developing nations like Nigeria.
{"title":"ESBL Production and Multidrug Resistance of Salmonella Serovars Isolates in Benue State","authors":"B. Okpa, G. Gberikon, C. Aguoru, I. O. Ogbonna","doi":"10.4236/ajmb.2020.103014","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103014","url":null,"abstract":"Studies on ESBL-producing and multi-drug resistance of Salmonella serovars distributed in Benue State were investigated. A total of four hundred and twenty (420) clinical stool samples, seventy (70) from each local government area were randomly collected from selected hospitals and analyzed for the presence of Salmonella spp. The isolates were characterized using Gram staining and biochemical tests. The result of AP120E biochemical test strip which contained dehydrated bacterial media and biochemical reagents in twenty (20) separate compartments. The result was obtained by evaluation of the compartments due to observed changes in after 24 hours where others were read by adding up reagents (Ferric chloride, Kovacs V.P reagents). The results were analyzed afterwards in accordance with the manufacturer’s software and positive results with ≥89% potential were confirmed as Salmonella spp. Amplified plasmids derived from 18 Salmonella strains recognized were made up of 23,130 base pairs. ESBL (Extended-spectrum beta-lactamases) genes were located on the plasmids. Two of the ESBL genes found were TEM genes and CTX-M (415 bp). Strains such as S. enterica Typhimurium-CP014981.1, S. enterica Enteritidis-CP007325.2, S. enterica Typhimurium-CP024619.1, S. enterica Typhimurium-CP023166.1, S. bongori-FR877557 and S. enterica Enteritidis-TY1 possessed TEM genes where as S. enterica Heidelberg-CP019176.1, S. enterica Typhi-AL513382.1 and S. enterica Typhimurium-MH196335.1 possessed CTX-M. Antibiotic resistance testing was performed using Kirby-Bauer disc diffusion method. The overall percentage susceptibility of the eight antibiotics tested on Salmonella serovars isolates shows that GEN had the highest % susceptibility of 100% followed by NIT (72.2%) and COT (66.7%) before and after plasmid curing. % susceptibility was lower before curing than after curing in CXC, CHL and TET. It was low (5.6%) in ERY while AUG recorded 0% susceptibility. Differences observed in curing status were insignificant (T = 0.33, P > 0.05). The presence of ESBL-producing and multi-drug resistant Salmonella serovars indicates an infection which presents a foremost peril to public health since such infections may be intricate to take care of and may consequently result in death of the infected patients. Constant periodic examination and prevention of drug abuse of antibiotics will assist in ensuring that this trend is curtailed especially in developing nations like Nigeria.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45243246","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 : 2020-02-17DOI: 10.4236/ajmb.2020.102008
S. Aslam, S. Khan, Aftab Ahmed, A. Dandekar
Cotton is considered as a major cash crop of the world. It earns huge foreign exchange by its valuable products; fiber, lint, cotton seed oil, hull and a lot more. Being an important fiber crop, it earns huge foreign exchange by contributing to textile and seed oil industry. This review summarizes cotton biology, its diversity and domestication, genome assembly, constraints in its production and methods to improve cotton plant to fulfill the need of textile and oil industry. But cotton is facing enormous biotic and abiotic stresses with insect pests being most prominent. Massive destruction caused by insects needs to be controlled for maintaining fruitful cotton crop production. Conventional breeding approaches are limited to improving single trait and integrate stable genes within plant genome in approximately 7 - 8 years. Improved biotechnological procedures have paved new pathways to target genes specifically and improve cotton germplasm in lesser time than conventional breeding.
{"title":"The Tale of Cotton Plant: From Wild Type to Domestication, Leading to Its Improvement by Genetic Transformation","authors":"S. Aslam, S. Khan, Aftab Ahmed, A. Dandekar","doi":"10.4236/ajmb.2020.102008","DOIUrl":"https://doi.org/10.4236/ajmb.2020.102008","url":null,"abstract":"Cotton is considered as a major cash crop of the world. It earns huge foreign exchange by its valuable products; fiber, lint, cotton seed oil, hull and a lot more. Being an important fiber crop, it earns huge foreign exchange by contributing to textile and seed oil industry. This review summarizes cotton biology, its diversity and domestication, genome assembly, constraints in its production and methods to improve cotton plant to fulfill the need of textile and oil industry. But cotton is facing enormous biotic and abiotic stresses with insect pests being most prominent. Massive destruction caused by insects needs to be controlled for maintaining fruitful cotton crop production. Conventional breeding approaches are limited to improving single trait and integrate stable genes within plant genome in approximately 7 - 8 years. Improved biotechnological procedures have paved new pathways to target genes specifically and improve cotton germplasm in lesser time than conventional breeding.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46384239","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 : 2020-01-10DOI: 10.4236/ajmb.2020.101007
E. J. Lee, S. Shin, S. Hyun, S. Kang
TRPV4 activity modulates cell activities including receptor trafficking and transcriptional or translational regulations. We tested its CRISPR/Cas9 scissor efficacy in HepG2 (HEK293) cell noticed that it worked well in both cell lines to eliminate TRPV4 genome sequences. To confirm TRPV4 functions in the cell morphology maintenance and cell growth (beyond Ca2+ channel), we compared its wound healing, cell surface area, survival property and soft agar growth ability after deletion of TRPV4 gene in the cells with its CRISPR/Cas9 system. With these experiments, we confirmed that TRPV4 is required not only to function as Ca2+ channel but also to maintain its proper cell morphology as a corner stone protein on the cell adhesion junction.
{"title":"Ablation of TRPV4 in HepG2 with Its CRISPR/Cas9 Enhances Its Wound Healing","authors":"E. J. Lee, S. Shin, S. Hyun, S. Kang","doi":"10.4236/ajmb.2020.101007","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101007","url":null,"abstract":"TRPV4 activity modulates cell activities including receptor trafficking and transcriptional or translational regulations. We tested its CRISPR/Cas9 scissor efficacy in HepG2 (HEK293) cell noticed that it worked well in both cell lines to eliminate TRPV4 genome sequences. To confirm TRPV4 functions in the cell morphology maintenance and cell growth (beyond Ca2+ channel), we compared its wound healing, cell surface area, survival property and soft agar growth ability after deletion of TRPV4 gene in the cells with its CRISPR/Cas9 system. With these experiments, we confirmed that TRPV4 is required not only to function as Ca2+ channel but also to maintain its proper cell morphology as a corner stone protein on the cell adhesion junction.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"10 1","pages":"74-89"},"PeriodicalIF":0.0,"publicationDate":"2020-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46971769","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 : 2020-01-01DOI: 10.4236/ajmb.2020.101005
J. Jin, Zhongjian Li, Lan-yong Zhao
Rosa rugosa is an important garden ornamental plant which belongs to the genus Rosa of the family Rosaceae. The current wild and cultivated R. rugosa are mostly purple, pink, a small amount of white, but lack of yellow, orange and so on. Flavonoids 3’-hydroxylase belongs to CYP75B subfamily of cytochrome P450, and is an essential enzyme in anthocyanins synthesis. In this experiment, RrF3’H gene was cloned from the petal of Rosa rugosa ‘Hunchun’ using RT-PCR, and bioinformatics analysis was performed. The RrF3’H gene’s full length of opening reading frame was 1687 bp, encoding 510 amino acids. The formulas of proteins encoded by RrF3’H were C2666H4149N699O734S24. The derived protein had a molecular weight of 58,506.95 Da. The aliphatic index was 90.94. It belongs to unstable hydrophilic protein. The protein consists of 46.76% α-helix, 31.04% random coil, 7.66% β-corner and 14.54% extended strand. The protein contains 21 Ser phosphorylation sites, 12 Thr phosphorylation sites, and 2 Tyr phosphorylation sites. The protein contained two O-glycosylation sites, located at positions 98 and 263 of the amino acid sequence respectively. The protein has a signal peptide site and a transmembrane structure. In addition, by comparing the expression levels of RrF3’H, we found RrF3’H was positively correlated with the depth of flower color.
{"title":"Cloning and Analysis of RrF3’ H in Rosa rugosa","authors":"J. Jin, Zhongjian Li, Lan-yong Zhao","doi":"10.4236/ajmb.2020.101005","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101005","url":null,"abstract":"Rosa rugosa is an important garden ornamental plant which belongs to the genus Rosa of the family Rosaceae. The current wild and cultivated R. rugosa are mostly purple, pink, a small amount of white, but lack of yellow, orange and so on. Flavonoids 3’-hydroxylase belongs to CYP75B subfamily of cytochrome P450, and is an essential enzyme in anthocyanins synthesis. In this experiment, RrF3’H gene was cloned from the petal of Rosa rugosa ‘Hunchun’ using RT-PCR, and bioinformatics analysis was performed. The RrF3’H gene’s full length of opening reading frame was 1687 bp, encoding 510 amino acids. The formulas of proteins encoded by RrF3’H were C2666H4149N699O734S24. The derived protein had a molecular weight of 58,506.95 Da. The aliphatic index was 90.94. It belongs to unstable hydrophilic protein. The protein consists of 46.76% α-helix, 31.04% random coil, 7.66% β-corner and 14.54% extended strand. The protein contains 21 Ser phosphorylation sites, 12 Thr phosphorylation sites, and 2 Tyr phosphorylation sites. The protein contained two O-glycosylation sites, located at positions 98 and 263 of the amino acid sequence respectively. The protein has a signal peptide site and a transmembrane structure. In addition, by comparing the expression levels of RrF3’H, we found RrF3’H was positively correlated with the depth of flower color.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513518","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 : 2020-01-01DOI: 10.4236/ajmb.2020.101001
I. Eto
Research Aims: Obesity and type 2 diabetes are known to be associated with increased risk of various types of cancer. However, the molecular biological mechanism of how the risk of cancer is increased in obesity or type 2 diabetes is not known. The aim this research is to investigate if the decreased expression of p27Kip1, a cell cycle repressor protein, plays a central role in this mechanism. Research Methods, Previous Studies and Theoretical Backgrounds: It is well known that the expression of p27Kip1 is increased by numerous nutritional or chemopreventive anti-cancer agents. But it has never been known that the expression of p27Kip1 could be decreased, rather than increased, and the risk of cancer could be increased, rather than decreased. This problem was solved recently and this new analytical method was used in this study. Results: 1) The expression of p27Kip1 was indeed significantly decreased in human obese type 2 diabetic individuals relative to the lean normal controls. 2) The expression of p27Kip1 was also significantly decreased in genetically obese rodents relative to the lean normal controls. Additionally, in obese rodents, the concentrations of glucose or insulin were significantly increased relative to the lean normal controls. 3) Using human cells cultured in vitro it was found that the increased concentrations of glucose or insulin decrease the expression of p27Kip1. Conclusions: These results suggest that higher concentrations of glucose or insulin increase the risk of various types of cancer in obesity or type 2 diabetes by decreasing the expression of p27Kip1.
{"title":"Higher Concentrations of Glucose or Insulin Increase the Risk of Various Types of Cancer in Obesity or Type 2 Diabetes by Decreasing the Expression of p27Kip1, a Cell Cycle Repressor Protein","authors":"I. Eto","doi":"10.4236/ajmb.2020.101001","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101001","url":null,"abstract":"Research Aims: Obesity and type 2 diabetes are known to be associated with increased risk of various types of cancer. However, the molecular biological mechanism of how the risk of cancer is increased in obesity or type 2 diabetes is not known. The aim this research is to investigate if the decreased expression of p27Kip1, a cell cycle repressor protein, plays a central role in this mechanism. Research Methods, Previous Studies and Theoretical Backgrounds: It is well known that the expression of p27Kip1 is increased by numerous nutritional or chemopreventive anti-cancer agents. But it has never been known that the expression of p27Kip1 could be decreased, rather than increased, and the risk of cancer could be increased, rather than decreased. This problem was solved recently and this new analytical method was used in this study. Results: 1) The expression of p27Kip1 was indeed significantly decreased in human obese type 2 diabetic individuals relative to the lean normal controls. 2) The expression of p27Kip1 was also significantly decreased in genetically obese rodents relative to the lean normal controls. Additionally, in obese rodents, the concentrations of glucose or insulin were significantly increased relative to the lean normal controls. 3) Using human cells cultured in vitro it was found that the increased concentrations of glucose or insulin decrease the expression of p27Kip1. Conclusions: These results suggest that higher concentrations of glucose or insulin increase the risk of various types of cancer in obesity or type 2 diabetes by decreasing the expression of p27Kip1.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"59 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513435","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 : 2020-01-01DOI: 10.4236/ajmb.2020.101003
T. Abiola, Ashimolowo O. Susan, B. Olusegun
Piroxicam is commonly used as anti-inflammatory and pain relieving drug; however, its side effects include fluid retention, renal damage and heart failure. This study aimed at evaluating the nephroprotective role of different doses of the tannin-rich extract of Chasmanthera dependens stem (TRECDS) on piroxicam-induced nephrotoxicity in adult male Wistar rats. Thirty-two adult rats were divided into four groups of eight rats per group and treated orally for ten days. Rats in group one received 0.5 ml normal saline (0.9% v/v) and served as normal control group. Rats in group two received 20 mg/kg body weight piroxicam alone. Rats in groups three and four received 20 mg/kg body weight of piroxicam with concomitant administration of 200 and 400 mg/kg body weight of TRECDS. At the expiration of the experiment, rats were sacrificed and the kidney was removed. Renal function was evaluated. The results showed that administration of piroxicam alone caused a significant elevation in the serum concentrations of albumin, creatinine, total protein, urea concentrations and the activity of renal nucleotidase with a reduction in the activity of glucose-6-phosphate dehydrogenase (G6PD) when compared to normal control (p < 0.05). Furthermore, renal tissue from the piroxicam alone treated group revealed a significant decrease in the activities of renal superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase as well as reduced glutathione with concomitant increase in lipid peroxidation and hydrogen peroxide generation. In addition, histological assessment of the renal tissue showed noticeable damage in piroxicam alone treated group. However, concomitant administration of TRECDS showed a dose-dependent reduction in the concentrations and the activity of the kidney markers with significant increase in the activities of G6PD and restores the antioxidant status of the kidney. The results show the nephroprotective potential of TREDS against piroxicam-induced renal damage.
{"title":"Tannin-Rich Extract of Chasmanthera dependens Stem Potential in Piroxicam-Induced Nephrotoxicity in Adult Male Wistar Rats","authors":"T. Abiola, Ashimolowo O. Susan, B. Olusegun","doi":"10.4236/ajmb.2020.101003","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101003","url":null,"abstract":"Piroxicam is commonly used as anti-inflammatory and pain relieving drug; however, its side effects include fluid retention, renal damage and heart failure. This study aimed at evaluating the nephroprotective role of different doses of the tannin-rich extract of Chasmanthera dependens stem (TRECDS) on piroxicam-induced nephrotoxicity in adult male Wistar rats. Thirty-two adult rats were divided into four groups of eight rats per group and treated orally for ten days. Rats in group one received 0.5 ml normal saline (0.9% v/v) and served as normal control group. Rats in group two received 20 mg/kg body weight piroxicam alone. Rats in groups three and four received 20 mg/kg body weight of piroxicam with concomitant administration of 200 and 400 mg/kg body weight of TRECDS. At the expiration of the experiment, rats were sacrificed and the kidney was removed. Renal function was evaluated. The results showed that administration of piroxicam alone caused a significant elevation in the serum concentrations of albumin, creatinine, total protein, urea concentrations and the activity of renal nucleotidase with a reduction in the activity of glucose-6-phosphate dehydrogenase (G6PD) when compared to normal control (p < 0.05). Furthermore, renal tissue from the piroxicam alone treated group revealed a significant decrease in the activities of renal superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase as well as reduced glutathione with concomitant increase in lipid peroxidation and hydrogen peroxide generation. In addition, histological assessment of the renal tissue showed noticeable damage in piroxicam alone treated group. However, concomitant administration of TRECDS showed a dose-dependent reduction in the concentrations and the activity of the kidney markers with significant increase in the activities of G6PD and restores the antioxidant status of the kidney. The results show the nephroprotective potential of TREDS against piroxicam-induced renal damage.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513475","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 : 2020-01-01DOI: 10.4236/ajmb.2020.101006
D. Funabara, Yoshinori Urakawa, D. Ishikawa, S. Kanoh
Troponin (Tn) is composed of three subunits (TnI, TnC and TnT) that bind Ca2+ and regulate striated muscle contraction in vertebrates. TnT’s function has been extensively described in vertebrates, but its role has been obscure in molluscan muscles. Our previous work indicated that the TnC and TnI subunits work in adductor phasic muscle, but not in catch muscle. Here, we have characterized TnT from the Japanese bivalve pearl oyster Pinctada fucata to start to explain the function of Tn in molluscan muscle contraction. We determined the primary structure of the full-length TnT protein from the P. fucata adductor muscle (Pifuc-TnT), and found that it is composed of 316 amino acid residues with a predicted molecular mass of 37.4 kDa. Multiple sequence alignment showed that Pifuc-TnT has an extension of >60 residues at the C-terminus that are not present in vertebrate TnTs, including known TnTs from other mollusks. Pifuc-TnT gene structure predictions using Splign alignment of the cDNA generated in this study and genome sequences indicated that Pifuc-TnT consists of 13 exons. Start and stop codons are located in exons 2 and 12, respectively. Quantitative real-time PCR revealed that the Pifuc-TnT gene was predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, slightly in gill, and not at all in mantle and foot. These findings suggest that TnT plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction. Isothermal titration calorimetry revealed that unlike vertebrate TnTs, Pifuc-TnT does not interact with P. fucata tropomyosin-1 nor with tropomyosin-2. These findings in P. fucata imply that Tn functions differently in molluscan muscle than it does in vertebrates.
{"title":"Troponin T from the Japanese Pearl Oyster Pinctada fucata: Molecular Cloning, Tissue Distribution, Gene Structure, and Interaction Analysis with Tropomyosin","authors":"D. Funabara, Yoshinori Urakawa, D. Ishikawa, S. Kanoh","doi":"10.4236/ajmb.2020.101006","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101006","url":null,"abstract":"Troponin (Tn) is composed of three subunits (TnI, TnC and TnT) that bind Ca2+ and regulate striated muscle contraction in vertebrates. TnT’s function has been extensively described in vertebrates, but its role has been obscure in molluscan muscles. Our previous work indicated that the TnC and TnI subunits work in adductor phasic muscle, but not in catch muscle. Here, we have characterized TnT from the Japanese bivalve pearl oyster Pinctada fucata to start to explain the function of Tn in molluscan muscle contraction. We determined the primary structure of the full-length TnT protein from the P. fucata adductor muscle (Pifuc-TnT), and found that it is composed of 316 amino acid residues with a predicted molecular mass of 37.4 kDa. Multiple sequence alignment showed that Pifuc-TnT has an extension of >60 residues at the C-terminus that are not present in vertebrate TnTs, including known TnTs from other mollusks. Pifuc-TnT gene structure predictions using Splign alignment of the cDNA generated in this study and genome sequences indicated that Pifuc-TnT consists of 13 exons. Start and stop codons are located in exons 2 and 12, respectively. Quantitative real-time PCR revealed that the Pifuc-TnT gene was predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, slightly in gill, and not at all in mantle and foot. These findings suggest that TnT plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction. Isothermal titration calorimetry revealed that unlike vertebrate TnTs, Pifuc-TnT does not interact with P. fucata tropomyosin-1 nor with tropomyosin-2. These findings in P. fucata imply that Tn functions differently in molluscan muscle than it does in vertebrates.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"10 1","pages":"61-73"},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513527","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 : 2020-01-01DOI: 10.4236/ajmb.2020.101002
S. Shin, E. J. Lee, S. Hyun, S. Kang
Serum- and glucocorticoid-induced kinase 1 (SGK1) is known to have consensus sequence of phosphorylation site R-x-R-x-x-(S/T)-Φ, where Φ is any hydrophobic amino acid and arginine residues are conserved at positions −5 and −3 relative to positions of Ser/Thr residues that are phosphorylated in the presence of SGK1. UNC-21-like kinase 2 (ULK2) also harbors putative SGK1 phosphorylation sites at both Ser507 (502RsRnsSG508) and Ser750 (745RtRttSV751) residues. Thus, the objective of this study was to determine whether Ser507 and Ser750 residues of ULK2 could be phosphorylation sites of SGK1 as one of its authentic substrate proteins. Using ULK2 507 and 750 serine residue un- or phosphorylation analog (S507AS750A or 507DS750D), we observed that modification of Ser507 or Ser750 residue was required to activate the kinase activity of ULK2 and sensitize ULK2 to stress or starvation while simultaneously enhancing its active state and autophagy characteristics, suggesting that phosphorylation at Ser750 or Ser507 residue could modulate its subcellular localization and protein interaction with AMPK1α to activate ULK2. We also observed that ULK2 autophagy activity was enhanced by GSK650394 (an SGK1 inhibitor) to compensate survival capacity through increasing its association with LC3 and phosphorylation. When SGK1 known to be associated with cell survival was inhibited by GSK650394, ULK2 autophagy pathway was activated to avoid cell death alternatively. Thus, our observations indicate that phosphorylation of ULK2 by SGK1 can regulate cell survival as an alternative modulation of ULK2 functions.
{"title":"SGK1 Inhibits ULK2 Autophagy Activity","authors":"S. Shin, E. J. Lee, S. Hyun, S. Kang","doi":"10.4236/ajmb.2020.101002","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101002","url":null,"abstract":"Serum- and glucocorticoid-induced kinase 1 (SGK1) is known to have consensus sequence of phosphorylation site R-x-R-x-x-(S/T)-Φ, where Φ is any hydrophobic amino acid and arginine residues are conserved at positions −5 and −3 relative to positions of Ser/Thr residues that are phosphorylated in the presence of SGK1. UNC-21-like kinase 2 (ULK2) also harbors putative SGK1 phosphorylation sites at both Ser507 (502RsRnsSG508) and Ser750 (745RtRttSV751) residues. Thus, the objective of this study was to determine whether Ser507 and Ser750 residues of ULK2 could be phosphorylation sites of SGK1 as one of its authentic substrate proteins. Using ULK2 507 and 750 serine residue un- or phosphorylation analog (S507AS750A or 507DS750D), we observed that modification of Ser507 or Ser750 residue was required to activate the kinase activity of ULK2 and sensitize ULK2 to stress or starvation while simultaneously enhancing its active state and autophagy characteristics, suggesting that phosphorylation at Ser750 or Ser507 residue could modulate its subcellular localization and protein interaction with AMPK1α to activate ULK2. We also observed that ULK2 autophagy activity was enhanced by GSK650394 (an SGK1 inhibitor) to compensate survival capacity through increasing its association with LC3 and phosphorylation. When SGK1 known to be associated with cell survival was inhibited by GSK650394, ULK2 autophagy pathway was activated to avoid cell death alternatively. Thus, our observations indicate that phosphorylation of ULK2 by SGK1 can regulate cell survival as an alternative modulation of ULK2 functions.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"531 1","pages":"12-28"},"PeriodicalIF":0.0,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513468","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}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：