Pub Date : 2020-01-01DOI: 10.4236/ajmb.2020.101004
Kishan Pavani Jankuti, Anusha Govindula, Rajkumar Jampala, M. A. Pasha, Aligeti Sravan Kumar, Sunayana Soorammagari, C. Satish, M. Ganesh
The main aim of this study is to determine the anti-hyerlipidemic and anti-obesity activity of Canephora robusta in hyperlipidemia induced rats. Prepared coffee bean extract (GCE) was procured from the market which is unroasted and contains more quantity of caffeine and chlorogenic acid when compared to roasted coffee. Male albino Wister rats are fed with high fat diet (HFD) for weeks to induce hyperlipidemia in rats, which are divided into 4 groups with 4 animals in each group. Test GCBE was given in doses of 200 mg/kg and 400 mg/kg to III and IV groups which are fed with HFD for 30 days. Then blood samples were collected through retro-orbital sinus by capillaries and serum is separated for analysis. The result obtained from lipid profile which includes total cholesterol, triglycerides, very low density lipoproteins, and low density lipoproteins shows the decreased level when compared to the hyperlipidemic control. This shows the significant reduction of total body weight (p < 0.05) when given with dose of 200 mg/kg and 400 mg/kg. The present study suggests that GCBE has anti-obesity and anti-hyperlipidemic activity, where 400 mg/kg is more effective to reduce the total body weight and lipid levels when compared to 200 mg/kg. Further studies on this extract may lead to identify the possible mechanism of action and isolation of active principle from the same.
{"title":"In-Vivo Anti-Hyperlipidemic Activity and Preliminary Phytochemical Screening of Canephora robusta","authors":"Kishan Pavani Jankuti, Anusha Govindula, Rajkumar Jampala, M. A. Pasha, Aligeti Sravan Kumar, Sunayana Soorammagari, C. Satish, M. Ganesh","doi":"10.4236/ajmb.2020.101004","DOIUrl":"https://doi.org/10.4236/ajmb.2020.101004","url":null,"abstract":"The main aim of this study is to determine the anti-hyerlipidemic and anti-obesity activity of Canephora robusta in hyperlipidemia induced rats. Prepared coffee bean extract (GCE) was procured from the market which is unroasted and contains more quantity of caffeine and chlorogenic acid when compared to roasted coffee. Male albino Wister rats are fed with high fat diet (HFD) for weeks to induce hyperlipidemia in rats, which are divided into 4 groups with 4 animals in each group. Test GCBE was given in doses of 200 mg/kg and 400 mg/kg to III and IV groups which are fed with HFD for 30 days. Then blood samples were collected through retro-orbital sinus by capillaries and serum is separated for analysis. The result obtained from lipid profile which includes total cholesterol, triglycerides, very low density lipoproteins, and low density lipoproteins shows the decreased level when compared to the hyperlipidemic control. This shows the significant reduction of total body weight (p < 0.05) when given with dose of 200 mg/kg and 400 mg/kg. The present study suggests that GCBE has anti-obesity and anti-hyperlipidemic activity, where 400 mg/kg is more effective to reduce the total body weight and lipid levels when compared to 200 mg/kg. Further studies on this extract may lead to identify the possible mechanism of action and isolation of active principle from the same.","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":"70513481","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}
Fauzun Shaona, R. Hassan, Sajib Chakraborty, S. Sultana, Jobaida Akther, A. Nabi
This study investigated distribution of HLA alleles (HLADRB1*01, *03, *04, *07, HLA-DQB1*0201, *0301/4) in 34 healthy controls and 57 rheumatoid arthritis (RA) patients in a Bangladeshi population and correlated the genotypic frequencies with clinical parameters. Frequency distribution of HLA-DRB1*04 (34%) and HLA-DRB1*01 (32%) were the highest followed by HLA-DQB1*0301/4 (29%) and HLA-DQB1*0201 (26%) in RA patients while HLA-DRB1*03 (12%) had lowest frequency. Plasma level of anti-CCP and rheumatoid factor antibodies confirmed diagnosis of RA patients that varied significantly between patients and healthy controls. Likewise, plasma levels of C-reactive protein, triglycerides, cholesterol, HDL-cholesterol and activities of AST and ALT exhibited significant variation between the two groups. In contrast, the levels of glucose, total protein, uric acid, LDL-cholesterol and plasma activity of ALP in RA patients had no significant deviations from healthy controls. Relationship between HLA genotype frequency and clinical parameters revealed that the mean levels of anti-CCP and rheumatoid factor antibodies were highest in the patients harboring HLA-DRB1*04 allele. These findings underpin the correlation between HLA genotype with clinical markers of RA which are indicative of disease severity. The positive correlation of these markers with certain HLA genes may be used to identify susceptible individuals who are likely to have RA in Bangladeshi population.
{"title":"Genotypic Analysis Revealed Association of HLA Alleles with Clinical Parameters in Bangladeshi Patients with Rheumatoid Arthritis","authors":"Fauzun Shaona, R. Hassan, Sajib Chakraborty, S. Sultana, Jobaida Akther, A. Nabi","doi":"10.4236/ajmb.2019.94013","DOIUrl":"https://doi.org/10.4236/ajmb.2019.94013","url":null,"abstract":"This study investigated distribution of HLA alleles (HLADRB1*01, *03, *04, *07, HLA-DQB1*0201, *0301/4) in 34 healthy controls and 57 rheumatoid arthritis (RA) patients in a Bangladeshi population and correlated the genotypic frequencies with clinical parameters. Frequency distribution of HLA-DRB1*04 (34%) and HLA-DRB1*01 (32%) were the highest followed by HLA-DQB1*0301/4 (29%) and HLA-DQB1*0201 (26%) in RA patients while HLA-DRB1*03 (12%) had lowest frequency. Plasma level of anti-CCP and rheumatoid factor antibodies confirmed diagnosis of RA patients that varied significantly between patients and healthy controls. Likewise, plasma levels of C-reactive protein, triglycerides, cholesterol, HDL-cholesterol and activities of AST and ALT exhibited significant variation between the two groups. In contrast, the levels of glucose, total protein, uric acid, LDL-cholesterol and plasma activity of ALP in RA patients had no significant deviations from healthy controls. Relationship between HLA genotype frequency and clinical parameters revealed that the mean levels of anti-CCP and rheumatoid factor antibodies were highest in the patients harboring HLA-DRB1*04 allele. These findings underpin the correlation between HLA genotype with clinical markers of RA which are indicative of disease severity. The positive correlation of these markers with certain HLA genes may be used to identify susceptible individuals who are likely to have RA in Bangladeshi population.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44198273","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}
Higher plants can adapt to abiotic stress to a certain degree. In this study, the impact of temperature stress on osmotic stress adapted and un-adapted cell lines of rice (Oryza sativa L.cv Swat-1) was observed. For the change in proline content, relative growth rate, saturated and unsaturated fatty acid were evaluated. The cell lines were incrementally adapted to 20% polyethylene glycol. The adapted lines showed significantly higher growth rate and proline content as compared to the un-adapted cell lines on temperature stress. Among saturated fatty acids palmitic acid (C16:0), stearic acid (C18:0) and myristic acid (C14:0) were the prominent fatty acids detected while among unsaturated fatty acid Oleic acid (C18:1c) and Linoleic acid (C18:2c) were the major fatty acids found. Under low temperature stress the percentage of saturated fatty acids was found to be lower (53%) in adapted cell line as compared to the un-adapted cell line (63%) while the percentage of saturation increased (83%) in adapted line under high temperature stress as compared to un-adapted line (70%). On the other hand at low temperature stress the percent level of unsaturated fatty acids in the adapted line was higher (48%) than the un-adapted cell line (37%). In conclusion, adaptation to one abiotic stress confers co-tolerance to the other abiotic stresses. Fatty acids saturation level could be a crucial factor in the plant ability to tolerate heat and cold stress.
{"title":"Fatty Acid Profiling of Polyethylene Glycol Adapted and Un-Adapted Cell Lines of Oryza sativa L.cv. Swat-1 under Temperature Stress","authors":"S. M. Shah, F. Ullah, Safdar Hussain Shah","doi":"10.4236/AJMB.2019.94011","DOIUrl":"https://doi.org/10.4236/AJMB.2019.94011","url":null,"abstract":"Higher plants can adapt to abiotic stress to a certain degree. In this study, the impact of temperature stress on osmotic stress adapted and un-adapted cell lines of rice (Oryza sativa L.cv Swat-1) was observed. For the change in proline content, relative growth rate, saturated and unsaturated fatty acid were evaluated. The cell lines were incrementally adapted to 20% polyethylene glycol. The adapted lines showed significantly higher growth rate and proline content as compared to the un-adapted cell lines on temperature stress. Among saturated fatty acids palmitic acid (C16:0), stearic acid (C18:0) and myristic acid (C14:0) were the prominent fatty acids detected while among unsaturated fatty acid Oleic acid (C18:1c) and Linoleic acid (C18:2c) were the major fatty acids found. Under low temperature stress the percentage of saturated fatty acids was found to be lower (53%) in adapted cell line as compared to the un-adapted cell line (63%) while the percentage of saturation increased (83%) in adapted line under high temperature stress as compared to un-adapted line (70%). On the other hand at low temperature stress the percent level of unsaturated fatty acids in the adapted line was higher (48%) than the un-adapted cell line (37%). In conclusion, adaptation to one abiotic stress confers co-tolerance to the other abiotic stresses. Fatty acids saturation level could be a crucial factor in the plant ability to tolerate heat and cold stress.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46784645","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}
Fawziya A. R. Ibrahim, Marwa S. Abouelenein, Ola A. Sakr, S. Ahmed, Ebtisam R. Zaher, S. E. E. Feky
Background: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a key player in the extrinsic pathway of apoptosis; it selectively damages cancer cells through binding to its surface receptors, however, cancers can escape this pathway through expression of dysfunctional decoy receptors. Purpose: The present study directed mainly to elucidate the serum TRAIL levels in breast cancer patients and to explore the variation in gene expression of TRAIL death and decoy receptors in breast cancer tissues, and to explore their role as prognostic markers in breast cancer as well as to detect their correlation with Patients’ Clinical Characteristics. Subjects and Methods: TRAIL levels were assayed in the sera of 124 breast cancer patients and 150 healthy females. Moreover, the expression of TRAIL death and decoy receptors was determined in both malignant and adjacent normal breast tissues collected from patients. ER, PR and Her-2 expression in breast cancer tissue were performed using immunohistochemical method. Apoptotic index (AI) was analyzed using HE in addition, serum TRAIL and profiling of TRAIL receptors expression may serve as prognostic markers in breast cancer patients.
{"title":"TRAIL and TRAIL Receptors as Prognostic Markers in Breast Cancer Patients","authors":"Fawziya A. R. Ibrahim, Marwa S. Abouelenein, Ola A. Sakr, S. Ahmed, Ebtisam R. Zaher, S. E. E. Feky","doi":"10.4236/ajmb.2019.94015","DOIUrl":"https://doi.org/10.4236/ajmb.2019.94015","url":null,"abstract":"Background: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a key player in the extrinsic pathway of apoptosis; it selectively damages cancer cells through binding to its surface receptors, however, cancers can escape this pathway through expression of dysfunctional decoy receptors. Purpose: The present study directed mainly to elucidate the serum TRAIL levels in breast cancer patients and to explore the variation in gene expression of TRAIL death and decoy receptors in breast cancer tissues, and to explore their role as prognostic markers in breast cancer as well as to detect their correlation with Patients’ Clinical Characteristics. Subjects and Methods: TRAIL levels were assayed in the sera of 124 breast cancer patients and 150 healthy females. Moreover, the expression of TRAIL death and decoy receptors was determined in both malignant and adjacent normal breast tissues collected from patients. ER, PR and Her-2 expression in breast cancer tissue were performed using immunohistochemical method. Apoptotic index (AI) was analyzed using HE in addition, serum TRAIL and profiling of TRAIL receptors expression may serve as prognostic markers in breast cancer patients.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44667814","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}
Hepatic injury has been reported to be associated with chloroquine therapy. Gomphrena celesioides has been claimed to have pleiotropic protective properties in the liver by traditional herbal practitioner but there is no scientific evidence to this claim. This investigation therefore sought to evaluate the effect of Gomphrena celesioides administration on chloroquine-induced hepatic injury in rats. Forty adult male rats were divided into five groups of eight rats and were treated orally once daily. Rats in group one received 1 ml/kg body weight of 0.9% normal saline; rats in group two received 250 mg/kg body weight of chloroquine for three days; groups three, four and five rats were pre-treated with 200 mg/kg, 400 mg/kg and 800 mg/kg body weight of methanol extract of Gomphrena celesiodes for three days and on the fourth day were given 250 mg/kg body weight of chloroquine for three days. The experiment lasted for seven days. Liver injury was manifested biochemically by a significant increase in serum level or activities of hepatic markers (aminotransferases, alkaline phosphate, bilirubin, cholesterol and gamma glutamyl transferase). In addition, hepatic tissue from chloroquine-treated rats showed a significant increase in lipid peroxidation with a decrease in hepatic superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and glutathione reservoirs. Moreover, the liver histopathologic evaluation revealed significant in chloroquine-treated rats. Gomphrena celesioides administration significantly alleviated chloroquine-induced pathologic changes in serum biochemistry and liver tissue. The results also suggest that Gomphrena celesioides possesses protective properties against chloroquine-induced liver injury via mitigation of drug-induced oxidative stress and its consequent events.
{"title":"Effect of Methanol Extract of Gomphrena celesioides on Chloroquine-Induced Hepatotoxicity and Oxidative Stress in Male Wistar Rats","authors":"T. Abiola, A. Dorcas, O. Babalola","doi":"10.4236/ajmb.2019.94014","DOIUrl":"https://doi.org/10.4236/ajmb.2019.94014","url":null,"abstract":"Hepatic injury has been reported to be associated with chloroquine therapy. Gomphrena celesioides has been claimed to have pleiotropic protective properties in the liver by traditional herbal practitioner but there is no scientific evidence to this claim. This investigation therefore sought to evaluate the effect of Gomphrena celesioides administration on chloroquine-induced hepatic injury in rats. Forty adult male rats were divided into five groups of eight rats and were treated orally once daily. Rats in group one received 1 ml/kg body weight of 0.9% normal saline; rats in group two received 250 mg/kg body weight of chloroquine for three days; groups three, four and five rats were pre-treated with 200 mg/kg, 400 mg/kg and 800 mg/kg body weight of methanol extract of Gomphrena celesiodes for three days and on the fourth day were given 250 mg/kg body weight of chloroquine for three days. The experiment lasted for seven days. Liver injury was manifested biochemically by a significant increase in serum level or activities of hepatic markers (aminotransferases, alkaline phosphate, bilirubin, cholesterol and gamma glutamyl transferase). In addition, hepatic tissue from chloroquine-treated rats showed a significant increase in lipid peroxidation with a decrease in hepatic superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and glutathione reservoirs. Moreover, the liver histopathologic evaluation revealed significant in chloroquine-treated rats. Gomphrena celesioides administration significantly alleviated chloroquine-induced pathologic changes in serum biochemistry and liver tissue. The results also suggest that Gomphrena celesioides possesses protective properties against chloroquine-induced liver injury via mitigation of drug-induced oxidative stress and its consequent events.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48422120","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}
Calponin is a basic actin-binding protein found widely in invertebrate tissues including catch muscle and therefore may participate in catch contraction. There is limited information about molluscan calponin and molecular characterization to reveal its function in the regulatory system. We previously identified and partially sequenced three calponin isoforms of the Japanese pearl oyster, Pinctada fucata (Pifuc-CP-1, Pifuc-CP-2 and Pifuc-CP-3). In this study, the full-length nucleotide sequences of the three isoforms were determined. The primary structures revealed that Pifuc-CP-1 consists of 324 amino acids (aa) with a molecular mass (Mw) of 34.7 kDa and an isoelectric point (pI) of 9.40. Pifuc-CP-2 is 303 aa in length with a Mw of 33.3 kDa and a pI of 9.30, and Pifuc-CP-3 is 398 aa in length with a Mw of 43.8 kDa and a pI of 8.55. Domain architecture prediction showed that the three isoforms have a single calponin homology (CH) domain and multiple calponin (CN) domains. Pifuc-CP-1, Pifuc-CP-2 and Pifuc-CP-3 possess four, three and five CN domains, respectively. Tissue distribution analysis indicated the presence of additional calponin isoforms and these isoforms are distributed widely in muscle and non-muscle tissues. Results of cDNA cloning revealed further four calponin isoforms: Pifuc-CP-4 (402 aa, 42.8 kDa, pI = 9.10), Pifuc-CP-5 (285 aa, 30.7 kDa, pI = 9.45), Pifuc-CP-6 (286 aa, 31.1 kDa, pI = 9.60) and Pifuc-CP-7 (302 aa, 33.3 kDa, pI = 9.10). The domain architecture of these four isoforms also consists of a single CH domain and multiple CN domains. Pifuc-CP-4 possesses six CN domains, whereas Pifuc-CP-5, Pifuc-CP-6 and Pifuc-CP-7 contain three CN domains. Sequence alignment of P. fucata calponin isoforms showed that Pifuc-CP-1, Pifuc-CP-2, Pifuc-CP-3 and Pifuc-CP-4 have identical CH domain sequences, whereas Pifuc-CP-5, Pifuc-CP-6 and Pifuc-CP-7 have identical CH domain sequences. The CN repeats were not well conserved. These findings suggest that P. fucata calponin isoforms function differently in each tissue.
{"title":"Calponin Isoform Expression in the Japanese Pearl Oyster, Pinctada fucata","authors":"D. Funabara, Y. Osakabe, S. Kanoh","doi":"10.4236/ajmb.2019.94012","DOIUrl":"https://doi.org/10.4236/ajmb.2019.94012","url":null,"abstract":"Calponin is a basic actin-binding protein found widely in invertebrate tissues including catch muscle and therefore may participate in catch contraction. There is limited information about molluscan calponin and molecular characterization to reveal its function in the regulatory system. We previously identified and partially sequenced three calponin isoforms of the Japanese pearl oyster, Pinctada fucata (Pifuc-CP-1, Pifuc-CP-2 and Pifuc-CP-3). In this study, the full-length nucleotide sequences of the three isoforms were determined. The primary structures revealed that Pifuc-CP-1 consists of 324 amino acids (aa) with a molecular mass (Mw) of 34.7 kDa and an isoelectric point (pI) of 9.40. Pifuc-CP-2 is 303 aa in length with a Mw of 33.3 kDa and a pI of 9.30, and Pifuc-CP-3 is 398 aa in length with a Mw of 43.8 kDa and a pI of 8.55. Domain architecture prediction showed that the three isoforms have a single calponin homology (CH) domain and multiple calponin (CN) domains. Pifuc-CP-1, Pifuc-CP-2 and Pifuc-CP-3 possess four, three and five CN domains, respectively. Tissue distribution analysis indicated the presence of additional calponin isoforms and these isoforms are distributed widely in muscle and non-muscle tissues. Results of cDNA cloning revealed further four calponin isoforms: Pifuc-CP-4 (402 aa, 42.8 kDa, pI = 9.10), Pifuc-CP-5 (285 aa, 30.7 kDa, pI = 9.45), Pifuc-CP-6 (286 aa, 31.1 kDa, pI = 9.60) and Pifuc-CP-7 (302 aa, 33.3 kDa, pI = 9.10). The domain architecture of these four isoforms also consists of a single CH domain and multiple CN domains. Pifuc-CP-4 possesses six CN domains, whereas Pifuc-CP-5, Pifuc-CP-6 and Pifuc-CP-7 contain three CN domains. Sequence alignment of P. fucata calponin isoforms showed that Pifuc-CP-1, Pifuc-CP-2, Pifuc-CP-3 and Pifuc-CP-4 have identical CH domain sequences, whereas Pifuc-CP-5, Pifuc-CP-6 and Pifuc-CP-7 have identical CH domain sequences. The CN repeats were not well conserved. These findings suggest that P. fucata calponin isoforms function differently in each tissue.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42807903","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}
Genome editing is considered as the most widely used approach of the present era. It had become a basic need of the current micro and molecular biological experiments. Gene engineering finds its widespread applications in medical, industry and agricultural sector. Unlike previous genetic engineering practices to insert or delete a part of genetic material at random place, genome editing allows the precise manipulation of DNA at a specific location. Zinc Finger Nucleases (ZFNs), Transcription Activator like Effector Nucleases (TALENs), Clustered Regularly Interspresed Short Palindromic repeats (CRISPR/Cas system) and meganucleases (recombinases) are the prime tools for editing an organism’s genome. Genome editing tools have an advantage to selectively delete or to integrate specific genes at specific loci. Use of recombinases for specifying site has further reduced time to integrate genes site specifically. Site specific gene stacking by the use of recombinases coupled with ZFNs, TALENs, or CRISPR/Cas genes have paved new pathways to target genes site specifically and to improve germplasm in lesser time than conventional breeding approaches.
{"title":"Genome Editing Tools: Need of the Current Era","authors":"S. Aslam, S. Khan, Aftab Ahmed, A. Dandekar","doi":"10.4236/AJMB.2019.93008","DOIUrl":"https://doi.org/10.4236/AJMB.2019.93008","url":null,"abstract":"Genome editing is considered as the most widely used approach of the present era. It had become a basic need of the current micro and molecular biological experiments. Gene engineering finds its widespread applications in medical, industry and agricultural sector. Unlike previous genetic engineering practices to insert or delete a part of genetic material at random place, genome editing allows the precise manipulation of DNA at a specific location. Zinc Finger Nucleases (ZFNs), Transcription Activator like Effector Nucleases (TALENs), Clustered Regularly Interspresed Short Palindromic repeats (CRISPR/Cas system) and meganucleases (recombinases) are the prime tools for editing an organism’s genome. Genome editing tools have an advantage to selectively delete or to integrate specific genes at specific loci. Use of recombinases for specifying site has further reduced time to integrate genes site specifically. Site specific gene stacking by the use of recombinases coupled with ZFNs, TALENs, or CRISPR/Cas genes have paved new pathways to target genes site specifically and to improve germplasm in lesser time than conventional breeding approaches.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44303565","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}
Nabil El Halawani, Manal El Sordi, Mona Aiad, B. E. Sabaa, Alia Hashim
Background: Members of the NFκB [p65] family have potential diagnostic and prognostic role in various inflammatory diseases and Lymphomas. Aim: We studied NFκB [p65] in paraffin blocks of hepatitis-C-virus [HCV] positive genotype-4 and HCV negative diffuse large B-cell lymphoma [DLBCL] patients, aiming at identification of its differential expression and prognosis in DLBCL and its subtypes; GCB and ABC. This is to establish its relation to HCV infection and its role in lymphogenesis. Besides assessing the role of new directly acting antiviral drugs [Sofusbuvir/Ledipasvir] concomitantly administered to [CHOP] combination in HCV positive DLBCL. Subjects and Methods: NFκB [p65] expression was assessed using Anti-NFκB [p65] antibody semi-quantitative technique in 30 newly diagnosed DLBCL patients [HCV positive [n = 15], HCV negative [n = 15]. Results: NFκB [p65] expression was higher in the HCV positive DLBCL patients than their HCV negative counterpart, with a positive correlation with the viral load [r = 0.536, p = 0.088]. NFκB [p65] expression was significantly more frequently detected in the ABC subtype than GCB subtype [p = 0.04]. Patients who expressed NFκB [p65] had higher incidence of extranodal involvement, advanced stages, higher LDH levels and IPI score. Besides, the expression of NFκB [P65] revealed an inferior overall response [OR] [p = 0.044]. Higher complete response rates to CHOP concomitantly with antiviral [ledipasvir/sofosbuvir] were encountered in the HCV positive group. In HCV positive group, NFκB [P65] displayed a positive relationship with the viral load and liver enzymes [p = 0.04], besides an inverse relation with serum albumin. This raises the possibility that NFκB [p65] expression is suggestive of the hepatic necro-inflammation in HCV patients. The ABC group presented more in advanced stages than GCB. Higher frequency of the ABC subgroup exhibited intermediate to high viral load, while it was less in the GCB. A statistically significant difference was found in the NFκB [p65] positive patients as regards MUM1 expression among the two groups [p ≤ 0.001]. Double positive [CD10+, MUM1+] and triple negative [CD10-, BCL6-, MUM1-] cases were encountered in the HCV positive group, and were characterized with a high NFκB [p65] expression. Conclusion: NFκB [p65] is expressed in patients with DLBCL, more frequently in ABC than in GCB subtypes. Expression of NFκB [p65] is associated with poor response to therapy in DLBCL. The NFκB [p65] disclosed an increased expression in HCV positive DLBCL compared to HCV negative group. The viral load displayed a positive correlation with the NFκB [p65] expression. Simultaneous administration of DAAs in combination with CHOP disclosed a better response and high tolerability.
{"title":"Diagnostic and Prognostic Value of Nuclear Factor Kappa-B in Diffuse Large B Cell Lymphoma in Egyptian Patients with Hepatitis C Virus Genotype 4","authors":"Nabil El Halawani, Manal El Sordi, Mona Aiad, B. E. Sabaa, Alia Hashim","doi":"10.4236/AJMB.2019.93010","DOIUrl":"https://doi.org/10.4236/AJMB.2019.93010","url":null,"abstract":"Background: Members of the NFκB [p65] family have potential diagnostic and prognostic role in various inflammatory diseases and Lymphomas. Aim: We studied NFκB [p65] in paraffin blocks of hepatitis-C-virus [HCV] positive genotype-4 and HCV negative diffuse large B-cell lymphoma [DLBCL] patients, aiming at identification of its differential expression and prognosis in DLBCL and its subtypes; GCB and ABC. This is to establish its relation to HCV infection and its role in lymphogenesis. Besides assessing the role of new directly acting antiviral drugs [Sofusbuvir/Ledipasvir] concomitantly administered to [CHOP] combination in HCV positive DLBCL. Subjects and Methods: NFκB [p65] expression was assessed using Anti-NFκB [p65] antibody semi-quantitative technique in 30 newly diagnosed DLBCL patients [HCV positive [n = 15], HCV negative [n = 15]. Results: NFκB [p65] expression was higher in the HCV positive DLBCL patients than their HCV negative counterpart, with a positive correlation with the viral load [r = 0.536, p = 0.088]. NFκB [p65] expression was significantly more frequently detected in the ABC subtype than GCB subtype [p = 0.04]. Patients who expressed NFκB [p65] had higher incidence of extranodal involvement, advanced stages, higher LDH levels and IPI score. Besides, the expression of NFκB [P65] revealed an inferior overall response [OR] [p = 0.044]. Higher complete response rates to CHOP concomitantly with antiviral [ledipasvir/sofosbuvir] were encountered in the HCV positive group. In HCV positive group, NFκB [P65] displayed a positive relationship with the viral load and liver enzymes [p = 0.04], besides an inverse relation with serum albumin. This raises the possibility that NFκB [p65] expression is suggestive of the hepatic necro-inflammation in HCV patients. The ABC group presented more in advanced stages than GCB. Higher frequency of the ABC subgroup exhibited intermediate to high viral load, while it was less in the GCB. A statistically significant difference was found in the NFκB [p65] positive patients as regards MUM1 expression among the two groups [p ≤ 0.001]. Double positive [CD10+, MUM1+] and triple negative [CD10-, BCL6-, MUM1-] cases were encountered in the HCV positive group, and were characterized with a high NFκB [p65] expression. Conclusion: NFκB [p65] is expressed in patients with DLBCL, more frequently in ABC than in GCB subtypes. Expression of NFκB [p65] is associated with poor response to therapy in DLBCL. The NFκB [p65] disclosed an increased expression in HCV positive DLBCL compared to HCV negative group. The viral load displayed a positive correlation with the NFκB [p65] expression. Simultaneous administration of DAAs in combination with CHOP disclosed a better response and high tolerability.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43080333","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}
Molluscan smooth muscles, such as the bivalve adductor muscles and the mussel anterior byssus retractor muscles (ABRM), exhibit a unique contraction called “catch”. Catch contraction is regulated through twitchin phosphorylation and dephosphorylation. Twitchin from the ABRM of the Mediterranean mussel, Mytilus galloprovincialis, is phosphorylated by cAMP-dependent protein kinase (PKA), and PKA phosphorylation sites are located in both the N- and C-terminal regions of the twitchin molecule. The D2 site, which is adjacently located to the C-terminus, participates in forming a myosin, actin, and twitchin complex that is thought to contribute towards the maintenance of tension in the catch state. In contrast, although it has been reported to interact with thin-filaments, the molecular function of the region including the D1 site has remained largely unstudied. Three additional PKA consensus sequences were identified near the D1 site; however, it was not known if these sites could be directly phosphorylated by PKA. Here, we performed phosphorylation assays to identify phosphorylation sites near the D1 site using recombinant protein variants (TWD1-SSSS, TWD1-AAAS, TWD1-AASA, TWD1-ASAA, TWD1-SAAA, and TWD1-AAAA). All variants, except TWD1-AAAA (where all phosphorylatable serine residues were replaced by alanines), were phosphorylated by PKA. The four phosphorylation sites were named D1-1, D1-2, D1-3, and D1-4 (the originally identified D1) in order from the N-terminus. Phosphorylation assays using a 1/12.5 weight ratio of PKA to each TWD1 variant revealed that D1-4 was the most rapidly phosphorylated, closely followed by D1-1. However, D1-2 and D1-3 were phosphorylated at a lower level under equivalent conditions and were not phosphorylated when PKA was incubated with each TWD1 variant at a 1/100 weight ratio. Furthermore, we observed that TWD1-SSSS was phosphorylated in a stepwise fashion. These findings contribute towards the elucidation of the function of the twitchin D1 region in the regulatory system of catch contraction.
{"title":"Phosphorylation Properties of the N-Terminal Region of Twitchin from Molluscan Catch Muscle","authors":"D. Funabara, Yuuki Nishimura, S. Kanoh","doi":"10.4236/AJMB.2019.93009","DOIUrl":"https://doi.org/10.4236/AJMB.2019.93009","url":null,"abstract":"Molluscan smooth muscles, such as the bivalve adductor muscles and the mussel anterior byssus retractor muscles (ABRM), exhibit a unique contraction called “catch”. Catch contraction is regulated through twitchin phosphorylation and dephosphorylation. Twitchin from the ABRM of the Mediterranean mussel, Mytilus galloprovincialis, is phosphorylated by cAMP-dependent protein kinase (PKA), and PKA phosphorylation sites are located in both the N- and C-terminal regions of the twitchin molecule. The D2 site, which is adjacently located to the C-terminus, participates in forming a myosin, actin, and twitchin complex that is thought to contribute towards the maintenance of tension in the catch state. In contrast, although it has been reported to interact with thin-filaments, the molecular function of the region including the D1 site has remained largely unstudied. Three additional PKA consensus sequences were identified near the D1 site; however, it was not known if these sites could be directly phosphorylated by PKA. Here, we performed phosphorylation assays to identify phosphorylation sites near the D1 site using recombinant protein variants (TWD1-SSSS, TWD1-AAAS, TWD1-AASA, TWD1-ASAA, TWD1-SAAA, and TWD1-AAAA). All variants, except TWD1-AAAA (where all phosphorylatable serine residues were replaced by alanines), were phosphorylated by PKA. The four phosphorylation sites were named D1-1, D1-2, D1-3, and D1-4 (the originally identified D1) in order from the N-terminus. Phosphorylation assays using a 1/12.5 weight ratio of PKA to each TWD1 variant revealed that D1-4 was the most rapidly phosphorylated, closely followed by D1-1. However, D1-2 and D1-3 were phosphorylated at a lower level under equivalent conditions and were not phosphorylated when PKA was incubated with each TWD1 variant at a 1/100 weight ratio. Furthermore, we observed that TWD1-SSSS was phosphorylated in a stepwise fashion. These findings contribute towards the elucidation of the function of the twitchin D1 region in the regulatory system of catch contraction.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48667700","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}
The genetic relationships among twenty-five accessions of Garcinia kola using six Random Amplified Polymorphic (RAPD) primers were evaluated in this study. The highest volume of total genomic DNA (2218/μl) was recorded in ON4 from Ikare, while the highest DNA concentration of 1.93 gl was found in OS3 from Ilesa. The highest Polymorphic Information Content (PIC) and gene diversity of 93.77% and 0.94 respectively were revealed by primer OPO2 compared to other primers. The dendogram generated from Unweighted Pair Group with Mean Average (UPGMA) clustering delineated two groups, A and B, consisting of 21 and 4 accessions respectively. This study clearly showed the level of molecular diversity in the accessions and the information provided could be utilized for genetic improvement and conservation of Garcinia kola.
{"title":"Molecular Evaluation of Garcinia kola Heckel Accessions Using RAPD Markers","authors":"O. Olawuyi, A. A. Azeez","doi":"10.4236/AJMB.2019.92004","DOIUrl":"https://doi.org/10.4236/AJMB.2019.92004","url":null,"abstract":"The genetic relationships among twenty-five accessions of Garcinia kola using six Random Amplified Polymorphic (RAPD) primers were evaluated in this study. The highest volume of total genomic DNA (2218/μl) was recorded in ON4 from Ikare, while the highest DNA concentration of 1.93 gl was found in OS3 from Ilesa. The highest Polymorphic Information Content (PIC) and gene diversity of 93.77% and 0.94 respectively were revealed by primer OPO2 compared to other primers. The dendogram generated from Unweighted Pair Group with Mean Average (UPGMA) clustering delineated two groups, A and B, consisting of 21 and 4 accessions respectively. This study clearly showed the level of molecular diversity in the accessions and the information provided could be utilized for genetic improvement and conservation of Garcinia kola.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42704445","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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