美国分子生物学期刊(英文)最新文献

{"title":"Specificity of Various Mitochondrial DNA (mtDNA), ND5, D-Loop, and Cty-b DNA Primers in Detecting Pig (Sus scrofa) DNA Fragments","authors":"J. Kusnadi, N. Ashari, E. L. Arumingtyas","doi":"10.4236/ajmb.2020.103010","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103010","url":null,"abstract":"Polymerase Chain Reaction (PCR) is an accurate, simple and fast analytical method. This technique is widely used in the identification of meat adulteration and meat-based processed food products. Three Mitochondrial DNA (mt-DNA) primers NADH Dehydrogenase sub unit 5 (ND5), D-Loop, and Cytochrome b (Cyt-b) were tested for their specificity in detecting of pig (Sus scrofa) DNA fragments. DNA genome from 6 meat samples (pork, beef, goat, lamb, and chicken) was amplified by PCR technique using three pairs of primers (ND5, D-Loop, and Cyt-b) and sequenced. The results of amplification using the three primers produced specific DNA bands with the lengths of 232 bp, 951 bp, and 404 bp, respectively. Comparison results with ND5, D-Loop, and Cyt-b gene sequences resulted in similarity values of 100%, 97%, and 99%, respectively. These showed that the mt-DNA primers of ND5, D-Loop, and Cyt-b genes can be recommended as specific primers in detecting pig (Sus scrofa) DNA fragments.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46475314","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}

{"title":"Lower Concentrations of Glucose or Insulin Decrease the Risk of Various Types of Cancer in the Long-Lived Ames Dwarf Mouse by Increasing the Expression of p27Kip1, a Cell-Cycle Repressor Protein","authors":"I. Eto","doi":"10.4236/ajmb.2020.103011","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103011","url":null,"abstract":"Introduction. The molecular biological mechanism of the increased \u0000incidence of the various types of cancer in obesity or type 2 diabetes in \u0000rodents or humans has largely been resolved in recent years. By contrast, the \u0000molecular biological mechanism of the decreased, not increased, incidence of \u0000the various types of cancer in the homozygous long-lived Ames dwarf mice still \u0000remains unresolved. Objective. The first objective of the present study \u0000was to investigate whether the decrease in the incidence of cancer in the homozygous \u0000long-lived Ames dwarf mice is due to the increase, not decrease, in the \u0000expression of p27Kip1, a cell cycle repressor protein. The second objective was \u0000to investigate whether the decrease in the incidence of cancer in the \u0000homozygous long-lived Ames dwarf mice is due to the decrease, not increase, in \u0000the levels of glucose or insulin. Methods. To achieve these objectives, \u0000we first performed western immunoblot analysis of the hepatic expression of \u0000p27Kip1 protein. We then performed, using a human breast cancer cell line in vitro, \u0000the luciferase reporter plasmid assay to determine whether the translation \u0000initiation activity of the p27Kip1 mRNA is increased when the concentrations of \u0000either glucose or insulin are decreased. Results and Conclusion. The \u0000results of the first objective indicated that the hepatic expression of p27Kip1 \u0000protein was up-regulated in the homozygous long-lived Ames dwarf mice as \u0000expected. We also found that the lower concentrations of glucose or insulin \u0000increased the translation initiation activity of the p27Kip1 mRNA.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70513533","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}

{"title":"PCR Based Detection and Phylogenetic Analysis of Fowl Adenovirus Strains Isolated from 2019 Epidemic from Punjab and Sindh, Pakistan","authors":"N. Sharif, M. Mehmood, S. Naqvi, Huma Anwar ul-Haq, S. S. Ahmed, M. Ghani, M. Shoaib, M. Hussain","doi":"10.4236/ajmb.2020.103016","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103016","url":null,"abstract":"Hydro-Pericardium Hepatitis (HPH) is an emerging infectious disease of commercial poultry, caused by different serotypes of Fowl Adeno Virus. The vertical transmission of the virus into the progeny may results in devastating damage, causing huge economic losses to its farmers. In present study, molecular typing is performed on basis of partially conserved hexon gene sequences, using a unique set of primers having common reverse oligo for simultaneous detection of FAdV1, FAdV-4 and FAdV-11. A total of 14 fowl adeno virus strains were isolated from 100 suspected adeno virus liver samples, collected from different districts in Pakistan, between 2018 and 2019. FASTA’s sequence alignment and phylogenetic analysis revealed that out of the 14, one isolate which belonged to group A showed 27% similarity with FAdV-1, while three isolates showed 99%, 95% & 45% similarity to FAdV-4 (Group C). Whereas, ten isolates showed more than 99% similarity to FAdV-11 (Group D). The serotypes FAdV1, FAdV-4 and FAdV-11 are prevailing in the breeder and broilers. These results hold great importance in rapid, reliable and simultaneous detection of the three FAdV serotypes. Therefore, fowl adeno virus vaccine production for commercial poultry shall be according to the prevalent field serotypes.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49564010","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}

{"title":"A Fluorescent Cell-Based Technique for Monitoring Efflux of MRP4","authors":"Tombari Pius Monsi, A. Ben-Chioma, Donaltus Onukwufor Onwuli","doi":"10.4236/ajmb.2020.103013","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103013","url":null,"abstract":"Background: Overexpression of efflux pumps is the drug resistance and adaptation mechanism employed by some eukaryotes and bacteria to transport endogenous and chemotherapeutic compounds from the intracellular to the extracellular environment. Aim: The study aimed at establishing a fluorescent cell-based assay to monitor the efflux activities of an ABC-transporter, multi-drug resistance protein 4 (MRP4). Methods: DH5α competent E. coli cells were transformed with pcDNA-MRP4 by the heat-shock process. The presence of the MRP4 gene was analyzed by the digestion of plasmid using EcoRI and analyzed on a 1% agarose gel. HEK 293 cells were transfected with purified pcDNA-MRP4 under optimized conditions using a Polyethylenimine (PEI) protocol. The level of MRP4 in the HEK 293 cells was characterized by western blotting analysis using M4I-10 anti-MRP4 and anti-Rat IgG (whole molecule)-Alkaline phosphatase antibodies. The fluorescent uptake study was performed by the incubation of 0.02 mM 8-[fluo-cAMP] with the MRP4-transfected and control HEK 293 cells for 1 h. The level of fluorescence was analyzed using fluorescence microscopy and spectrometer. Results: The agarose gel analysis showed a plasmid of 9.4 kb and restriction product of 5 kb, which correspond with the pcDNA and MRP4 sizes respectively. The western blot results of the transfection showed 4 μg pcDNA-MRP4 and the N/P ratio of 9 was the optimized condition to transfect our HEK 293 cells as it showed the broadest band. In the efflux studies, the fluorescence images of the MRP4-transfected HEK 293 cells were very low compared to the untransfected control. The level of fluorescence accumulation was significantly (P ≤ 0.0001) higher 228.6 ± 13.1 RFU in the untransfected cells than the MRP4-transfected cells 8.6 ± 1.8 RFU. Conclusion: The higher levels of fluorescence detected in the control in both the fluorescent microscopy and spectrophotometer showed that MRP4-transfected cells had effluxed the 8-[fluo-cAMP] substrate out of the cell. This method could be employed in the detection of MRP4 functions in bacteria and cancer cells.","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43000818","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}

{"title":"Evaluation of HBsAg Quantification as Surrogate to HBV DNA Viral Load in Hepatitis B Infected Patients in Anambra State, Nigeria","authors":"Chinwe Obiomah, G. Amilo, Israel Ndulue","doi":"10.4236/ajmb.2020.103009","DOIUrl":"https://doi.org/10.4236/ajmb.2020.103009","url":null,"abstract":"Hepatitis B is an infectious disease of great public health importance. Nigeria \u0000is one of the countries with the highest incidence of Hepatitis B Virus (HBV) \u0000infection worldwide. However, the accessibility and affordability of HBV \u0000DNA quantification (viral load) assay is the key laboratory test for therapy initiation, \u0000and monitoring is a challenge to HBV management. This study \u0000aimed at determining the relationship between HBV DNA quantification \u0000and routine haemato-serological parameters in order to develop a more \u0000cost-effective diagnostic algorithm for Hepatitis B management. Cross \u0000sectional study design was used with a total of 264 subjects comprising of 88 \u0000HBsAg seropositive treatment naïve subjects, 88 HBsAg seropositive subjects \u0000on antiviral therapy as case subjects and 88 age-matched apparently healthy \u0000HBsAg seronegative individuals were recruited as control subjects. Hepatitis \u0000B Virus DNA assay was performed using real time PCR technique while \u0000ELISA technique was used for Hepatitis B surface antigen quantification. \u0000HBsAg quantification showed strong positive correlation with HBV DNA \u0000viral load both in treatment and non-treatment groups (r = 0.673; p","PeriodicalId":65391,"journal":{"name":"美国分子生物学期刊(英文)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42814861","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}

{"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":null,"pages":null},"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}

{"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":null,"pages":null},"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}

{"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":null,"pages":null},"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}

{"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":null,"pages":null},"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}

{"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":null,"pages":null},"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}