Pub Date : 2022-09-19DOI: 10.35118/apjmbb.2022.030.3.12
M. Ibahim, Narimah Abdul Hamid Hasani, Fatimah Sham, E. Omar, Syed Baharom Syed Ahmad Fuad, Muhammad Khalis Abdul Karim, N. Hasan
We presented a flow cytometry protocol to isolate and measure the cellular component of tumour immune microenvironment (TIME) of mice bearing breast cancer models. The immune cells infiltrating the tumour microenvironment (TME) have been getting more attention as the determination of their pro- or/and anti-tumorigenic activity contributes to cancer treatment success. In addition, our group has a strong interest to investigate the infiltration of eosinophils within the TME as their tumorigenic role is still unclear. This study aims to measure the immune cells infiltrating the TME, including eosinophils using flow cytometry with 6-colour detection. The single-cell suspensions derived from tumour sections of mice bearing EMT6 tumour model were harvested (n=4) and treated with CD45, Siglec-F, I-A/I-E, CD 11b, and Ly6G antibodies. A gating protocol was used based on the reported publications. Results showed that immune cells in the tumour section were detected by positive staining to CD45. Neutrophils were isolated based on Ly6G+, while the remaining granulocytes, such as macrophage and dendritic cells (DC), were isolated based on I-A/I-E+SCChi, and eosinophils on the positive expression of Siglec F. Overall, our tumour model presented with the highest percentage of neutrophils (63.05 ± 5.61%), while eosinophils constitute 1.64 ± 0.75% of the total population. In conclusion, our flow cytometric protocol with 6 colour detection are able to isolate and measure several immune cells within the TME, including eosinophils from the tumour section.
{"title":"Development of flow cytometry analysis on measuring tumour immune microenvironment (TIME) in mice bearing EMT6 tumour model","authors":"M. Ibahim, Narimah Abdul Hamid Hasani, Fatimah Sham, E. Omar, Syed Baharom Syed Ahmad Fuad, Muhammad Khalis Abdul Karim, N. Hasan","doi":"10.35118/apjmbb.2022.030.3.12","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.12","url":null,"abstract":"We presented a flow cytometry protocol to isolate and measure the cellular component of tumour immune microenvironment (TIME) of mice bearing breast cancer models. The immune cells infiltrating the tumour microenvironment (TME) have been getting more attention as the determination of their pro- or/and anti-tumorigenic activity contributes to cancer treatment success. In addition, our group has a strong interest to investigate the infiltration of eosinophils within the TME as their tumorigenic role is still unclear. This study aims to measure the immune cells infiltrating the TME, including eosinophils using flow cytometry with 6-colour detection. The single-cell suspensions derived from tumour sections of mice bearing EMT6 tumour model were harvested (n=4) and treated with CD45, Siglec-F, I-A/I-E, CD 11b, and Ly6G antibodies. A gating protocol was used based on the reported publications. Results showed that immune cells in the tumour section were detected by positive staining to CD45. Neutrophils were isolated based on Ly6G+, while the remaining granulocytes, such as macrophage and dendritic cells (DC), were isolated based on I-A/I-E+SCChi, and eosinophils on the positive expression of Siglec F. Overall, our tumour model presented with the highest percentage of neutrophils (63.05 ± 5.61%), while eosinophils constitute 1.64 ± 0.75% of the total population. In conclusion, our flow cytometric protocol with 6 colour detection are able to isolate and measure several immune cells within the TME, including eosinophils from the tumour section.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80743103","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 : 2022-09-08DOI: 10.35118/apjmbb.2022.030.3.11
N. A. Sharudin, Nur Amira Khairil Anwar, Muhamad Najmi Mohd Nazri, Ahmad Hafiz Murtadha, Fatin Hamimi Hamat@Mustafa, M. Sarmiento, A. Acosta, N. Yaacob, N. F. Mokhtar
‘Neonatal’ Nav1.5 (nNav1.5) is a potent tumour metastasis marker found especially in aggressive human breast cancer cells in vitro, in tumour tissues of in vivo metastatic animal models and in patients positive for lymph-node metastasis. Its expression has been recently described in human brain neuroblastoma and astrocytoma. However, a thorough understanding of nNav1.5’s role in cancers has been limited by the lack of specific antibodies against it. Here, a mouse monoclonal antibody, 4H8 mAb-nNav1.5, was obtained and characterised concerning its efficacy in detecting nNav1.5 using indirect ELISA, surface plasmon resonance (SPR), Western blotting and immunofluorescence microscopy. 4H8 mAb-nNav1.5 was selected from a panel of hybridoma clones raised against nNav1.5 specific peptide (15 mers). The antibody exhibited linear association against nNav1.5 specific-linear peptide in indirect ELISA and was supported by SPR. The antibody also demonstrated strong immunoreactivity in immunofluorescence imaging of nNav1.5-abundant cells, human and mouse aggressive breast cancer cells, MDA-MB-231 and 4T1, respectively, which was not observed in nNav1.5-deficient cells, human less aggressive breast cancer cells, MCF-7 and non-cancerous breast epithelial cells, MCF-10A. This study demonstrates the initial description of 4H8 mAb-nNav1.5, which could serve as a beneficial tool to enhance future studies on nNav1.5 expression and function in cancers.
{"title":"Development and characterisation of mouse monoclonal antibody against ‘neonatal’ Nav1.5","authors":"N. A. Sharudin, Nur Amira Khairil Anwar, Muhamad Najmi Mohd Nazri, Ahmad Hafiz Murtadha, Fatin Hamimi Hamat@Mustafa, M. Sarmiento, A. Acosta, N. Yaacob, N. F. Mokhtar","doi":"10.35118/apjmbb.2022.030.3.11","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.11","url":null,"abstract":"‘Neonatal’ Nav1.5 (nNav1.5) is a potent tumour metastasis marker found especially in aggressive human breast cancer cells in vitro, in tumour tissues of in vivo metastatic animal models and in patients positive for lymph-node metastasis. Its expression has been recently described in human brain neuroblastoma and astrocytoma. However, a thorough understanding of nNav1.5’s role in cancers has been limited by the lack of specific antibodies against it. Here, a mouse monoclonal antibody, 4H8 mAb-nNav1.5, was obtained and characterised concerning its efficacy in detecting nNav1.5 using indirect ELISA, surface plasmon resonance (SPR), Western blotting and immunofluorescence microscopy. 4H8 mAb-nNav1.5 was selected from a panel of hybridoma clones raised against nNav1.5 specific peptide (15 mers). The antibody exhibited linear association against nNav1.5 specific-linear peptide in indirect ELISA and was supported by SPR. The antibody also demonstrated strong immunoreactivity in immunofluorescence imaging of nNav1.5-abundant cells, human and mouse aggressive breast cancer cells, MDA-MB-231 and 4T1, respectively, which was not observed in nNav1.5-deficient cells, human less aggressive breast cancer cells, MCF-7 and non-cancerous breast epithelial cells, MCF-10A. This study demonstrates the initial description of 4H8 mAb-nNav1.5, which could serve as a beneficial tool to enhance future studies on nNav1.5 expression and function in cancers.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88475071","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 : 2022-08-23DOI: 10.35118/apjmbb.2022.030.3.09
C. Lim, F. Wong, Joe-Chien Lim, Liza-Pilomina Xavier, W. Goh
Somaclonal variations in banana tissue culture is not uncommon. In our tissue culture laboratory, the tissue-cultured Lakatan variety exhibited abnormalities (i.e., malformed bunches/ fingers, stunted growth or sterility) that affect the fruit yield. Using a methylation-sensitive inter-simple sequence repeats (ISSR) approach, the genomic DNA digested with methylation-sensitive restriction enzyme MspI of 274 tissue-cultured Lakatan plants were screened. The ISSR profile of the abnormal samples were identical but different from the normal samples by the absence of 997 bp band in the abnormal samples. The sensitivity of the marker employed in this study is estimated to be 93.4%. We suggest that this abnormality in the tissue-cultured Lakatan is a result of epigenetic changes which could have been induced in the banana tissue culture process. Our study also showed that this abnormality can be transmitted to the next generation through asexual propagation. The methylation-sensitive ISSR method adopted in this study is a promising tool for early detection of such abnormality in the micropropagated Lakatan banana plants.
{"title":"Early detection of abnormality in the micropropagated Lakatan banana plants using methylation-sensitive ISSR","authors":"C. Lim, F. Wong, Joe-Chien Lim, Liza-Pilomina Xavier, W. Goh","doi":"10.35118/apjmbb.2022.030.3.09","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.09","url":null,"abstract":"Somaclonal variations in banana tissue culture is not uncommon. In our tissue culture laboratory, the tissue-cultured Lakatan variety exhibited abnormalities (i.e., malformed bunches/ fingers, stunted growth or sterility) that affect the fruit yield. Using a methylation-sensitive inter-simple sequence repeats (ISSR) approach, the genomic DNA digested with methylation-sensitive restriction enzyme MspI of 274 tissue-cultured Lakatan plants were screened. The ISSR profile of the abnormal samples were identical but different from the normal samples by the absence of 997 bp band in the abnormal samples. The sensitivity of the marker employed in this study is estimated to be 93.4%. We suggest that this abnormality in the tissue-cultured Lakatan is a result of epigenetic changes which could have been induced in the banana tissue culture process. Our study also showed that this abnormality can be transmitted to the next generation through asexual propagation. The methylation-sensitive ISSR method adopted in this study is a promising tool for early detection of such abnormality in the micropropagated Lakatan banana plants.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86106814","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 : 2022-08-15DOI: 10.35118/apjmbb.2022.030.3.08
Ianne Kong, Zi Wen Heng, L. Pui
Edible film is commonly applied on meat products. In this study, kesum leaf extract (KE) and curry leaf extract (CLE) were incorporated into chitosan based film, in varying concentrations of KE and CLE to determine the physicochemical, mechanical and antimicrobial properties of chitosan film formed. Therefore, the purpose of this research was to assess and compare the effect of chitosan film incorporated with CLE and KE on microbiological properties of chicken breast meat. Increase in KE and CLE concentrations leads to the decrease in mechanical strength of the chitosan film, while increased in its water solubility. In addition, the addition of KE and CLE to the chitosan films produced film that is darker, greener and less yellowish in colour compared to plain films. Increased KE and CLE concentrations increase the inhibition zone against Staphylococcus aureus and Salmonella spp. in terms of antimicrobial activity. In storage test, control and sample (chicken breast meat wrapped with chitosan film incorporated with 2.0% (w/v) KE and 2.0% (w/v) CLE) were kept at 4°C for 14 days. The sample were reported to have pH values, mesophilic and psychrotrophic total plate counts that were considerably lower than control during the storage time. The sample coated with chitosan film incorporated with 2.0% (w/v) of KE and 2.0% (w/v) CLE improved the storage life of chicken breast meat by at least 2 days. Chitosan films incorporated with KE and CLE held enormous promise as active packaging materials for meat preservation.
{"title":"Development of chitosan edible film incorporated with curry leaf and kesum for the packaging of chicken breast meat","authors":"Ianne Kong, Zi Wen Heng, L. Pui","doi":"10.35118/apjmbb.2022.030.3.08","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.08","url":null,"abstract":"Edible film is commonly applied on meat products. In this study, kesum leaf extract (KE) and curry leaf extract (CLE) were incorporated into chitosan based film, in varying concentrations of KE and CLE to determine the physicochemical, mechanical and antimicrobial properties of chitosan film formed. Therefore, the purpose of this research was to assess and compare the effect of chitosan film incorporated with CLE and KE on microbiological properties of chicken breast meat. Increase in KE and CLE concentrations leads to the decrease in mechanical strength of the chitosan film, while increased in its water solubility. In addition, the addition of KE and CLE to the chitosan films produced film that is darker, greener and less yellowish in colour compared to plain films. Increased KE and CLE concentrations increase the inhibition zone against Staphylococcus aureus and Salmonella spp. in terms of antimicrobial activity. In storage test, control and sample (chicken breast meat wrapped with chitosan film incorporated with 2.0% (w/v) KE and 2.0% (w/v) CLE) were kept at 4°C for 14 days. The sample were reported to have pH values, mesophilic and psychrotrophic total plate counts that were considerably lower than control during the storage time. The sample coated with chitosan film incorporated with 2.0% (w/v) of KE and 2.0% (w/v) CLE improved the storage life of chicken breast meat by at least 2 days. Chitosan films incorporated with KE and CLE held enormous promise as active packaging materials for meat preservation.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83311325","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 : 2022-08-08DOI: 10.35118/apjmbb.2022.030.3.07
Heena Bisht, Narayan Kumar
As we progress farther into the industrial age of the twenty-first century, we see that many types of pollutants emitted into the air, water, and land are becoming increasingly burdensome to our environment. These pollutants have a major impact on humans, plants, and animals. Heavy metals are the most hazardous elements in our ecosystem since they are extremely harmful to the environment and continue to build up in our food chain. We must learn about the harmful consequences of heavy metals and work to reduce them using the most environmentally friendly methods feasible. This review discusses how bioremediation helps to reduce heavy metal concentrations in our ecosystem using biological agents, such as algae, fungi, bacteria, and plants. The paper also investigates various phytoremediation and microbial remediation mechanisms involved in metal detoxification or transformation into less toxic forms, which lower the adverse effects of heavy metals in animals, plants, and humans.
{"title":"Plant and microbe mediated bioremediation: A long-term remedy for heavy metal pollution","authors":"Heena Bisht, Narayan Kumar","doi":"10.35118/apjmbb.2022.030.3.07","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.07","url":null,"abstract":"As we progress farther into the industrial age of the twenty-first century, we see that many types of pollutants emitted into the air, water, and land are becoming increasingly burdensome to our environment. These pollutants have a major impact on humans, plants, and animals. Heavy metals are the most hazardous elements in our ecosystem since they are extremely harmful to the environment and continue to build up in our food chain. We must learn about the harmful consequences of heavy metals and work to reduce them using the most environmentally friendly methods feasible. This review discusses how bioremediation helps to reduce heavy metal concentrations in our ecosystem using biological agents, such as algae, fungi, bacteria, and plants. The paper also investigates various phytoremediation and microbial remediation mechanisms involved in metal detoxification or transformation into less toxic forms, which lower the adverse effects of heavy metals in animals, plants, and humans.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84736182","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 : 2022-07-25DOI: 10.35118/apjmbb.2022.030.3.04
Farrah Hazwani, Indah Mohd Amin, Mohd Aizat Abdul Rahim
90% - 95% of oral malignancies are reported to be contributed by oral squamous cell carcinoma (OSCC). Patients with oral cancer also have suffered from a stagnant survival rate of 50% for decades. With the advancement in technologies and medicine discovery, the standard solution for oral cancer is via chemotherapy and surgery. An alternative for the treatment is by incorporating a genotoxic drug, cisplatin. Unfortunately, cisplatin has been reported by many patients to give unpleasant effects such as nausea, vomiting, diarrhea and abdominal pain. Hence, a more reliable treatment needs to be unleashed. Allicin (diallylthiosulfinate) is well known for its antioxidant, antifungal, anti-inflammatory and antimicrobial purposes. Numerous studies have also reported the effectiveness of allicin as an anticancer agent against multiple cancer cell strains. With treatment of allicin in a dose-dependent manner, inhibition of carcinoma cells proliferation through a programmed cell death called apoptosis was considered favourable. Allicin induces apoptosis by activating cascades of caspases and caspase independent pathways as well as increasing p53 and Bax/Bcl2 expression. Allicin serves many potentials to fight off oral cancer carcinoma cells but there seems to be a limited amount of study in the oral cancer area. Therefore, this review article would like to highlight the reported studies on allicin in various cancer cell lines including oral cancer cells.
{"title":"Molecular mechanism of allicin-induced apoptosis in human oral squamous cell carcinoma (OSCC)","authors":"Farrah Hazwani, Indah Mohd Amin, Mohd Aizat Abdul Rahim","doi":"10.35118/apjmbb.2022.030.3.04","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.04","url":null,"abstract":"90% - 95% of oral malignancies are reported to be contributed by oral squamous cell carcinoma (OSCC). Patients with oral cancer also have suffered from a stagnant survival rate of 50% for decades. With the advancement in technologies and medicine discovery, the standard solution for oral cancer is via chemotherapy and surgery. An alternative for the treatment is by incorporating a genotoxic drug, cisplatin. Unfortunately, cisplatin has been reported by many patients to give unpleasant effects such as nausea, vomiting, diarrhea and abdominal pain. Hence, a more reliable treatment needs to be unleashed. Allicin (diallylthiosulfinate) is well known for its antioxidant, antifungal, anti-inflammatory and antimicrobial purposes. Numerous studies have also reported the effectiveness of allicin as an anticancer agent against multiple cancer cell strains. With treatment of allicin in a dose-dependent manner, inhibition of carcinoma cells proliferation through a programmed cell death called apoptosis was considered favourable. Allicin induces apoptosis by activating cascades of caspases and caspase independent pathways as well as increasing p53 and Bax/Bcl2 expression. Allicin serves many potentials to fight off oral cancer carcinoma cells but there seems to be a limited amount of study in the oral cancer area. Therefore, this review article would like to highlight the reported studies on allicin in various cancer cell lines including oral cancer cells.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84576988","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 : 2022-07-25DOI: 10.35118/apjmbb.2022.030.3.06
Lailly Nur Uswatul Hasanah, Dewi Masuroh, Ika Wahyuni, R. Oktarianti, S. Wathon, A. Labes, E. Sulistyaningsih, K. Senjarini
Since the malaria outbreak in 2011, the breeding place of Anopheles in Bangsring Village on Banyuwangi District has been monitored by District Public Health Office as part of a vector surveillance program. Morphological identification is still a standard tool to observe Anopheles occurrence and diversity, but the presence of cryptic species made it unreliable. In this study, a molecular approach called DNA barcoding technique was used to assist the morphology-based techniques to identify Anopheles species found in Bangsring. The internal transcribed spacer 2 (ITS2) sequence was used as molecular marker. Based on the morphological features, we were able to identify Anopheles (An.) vagus, An. subpictus, An. sundaicus and An. aconitus. ITS2 sequences from the four identified species were then analyzed simultaneously with eighteen reference sequences from NCBI which had a high similarity of 98-100%. The NJ phylogenetic tree formed three major clades, where the two clades as monophyletic clades were An. vagus and An. aconitus. Another clade was formed as polyphyletic clade containing An. subpictus and An. sundaicus. Although An. subpictus and An. sundaicus were placed in the same clade, seven nucleotide differences were observed in their ITS2 sequence. The intra-specific variation of those two species was 0.08 and 0.49%, respectively, while the interspecific variation was 1.39%. Interspecific variation which was higher than the mean intra-specific variation might indicate that An. sundaicus and An. subpictus were a distantly species. However, the value of interspecific variation lower than 3% might also indicate that those species were classified as a complex species. All ITS2 sequences from morphologically identified species had similar results with molecular-based techniques. This result showed that molecular identification using the ITS2 sequence was reliable in supporting morphological identification among closely related anopheline mosquitoes and gave further information about their evolutionary divergence.
{"title":"Internal transcribed spacer 2 (ITS2) based molecular identification of malaria vectors from Bangsring Banyuwangi-Indonesia","authors":"Lailly Nur Uswatul Hasanah, Dewi Masuroh, Ika Wahyuni, R. Oktarianti, S. Wathon, A. Labes, E. Sulistyaningsih, K. Senjarini","doi":"10.35118/apjmbb.2022.030.3.06","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.06","url":null,"abstract":"Since the malaria outbreak in 2011, the breeding place of Anopheles in Bangsring Village on Banyuwangi District has been monitored by District Public Health Office as part of a vector surveillance program. Morphological identification is still a standard tool to observe Anopheles occurrence and diversity, but the presence of cryptic species made it unreliable. In this study, a molecular approach called DNA barcoding technique was used to assist the morphology-based techniques to identify Anopheles species found in Bangsring. The internal transcribed spacer 2 (ITS2) sequence was used as molecular marker. Based on the morphological features, we were able to identify Anopheles (An.) vagus, An. subpictus, An. sundaicus and An. aconitus. ITS2 sequences from the four identified species were then analyzed simultaneously with eighteen reference sequences from NCBI which had a high similarity of 98-100%. The NJ phylogenetic tree formed three major clades, where the two clades as monophyletic clades were An. vagus and An. aconitus. Another clade was formed as polyphyletic clade containing An. subpictus and An. sundaicus. Although An. subpictus and An. sundaicus were placed in the same clade, seven nucleotide differences were observed in their ITS2 sequence. The intra-specific variation of those two species was 0.08 and 0.49%, respectively, while the interspecific variation was 1.39%. Interspecific variation which was higher than the mean intra-specific variation might indicate that An. sundaicus and An. subpictus were a distantly species. However, the value of interspecific variation lower than 3% might also indicate that those species were classified as a complex species. All ITS2 sequences from morphologically identified species had similar results with molecular-based techniques. This result showed that molecular identification using the ITS2 sequence was reliable in supporting morphological identification among closely related anopheline mosquitoes and gave further information about their evolutionary divergence.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80046337","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 : 2022-07-25DOI: 10.35118/apjmbb.2022.030.3.03
Yuh Leng Teo, Shu Ting Chang, W. K. Toh, C. Ho, P. Loh, H. Wong
Tumor-inducing (Ti) plasmid is the requisite for Agrobacterium-mediated plant transformation. Over decades, continuous efforts have been made to improve the efficiency of Agrobacterium-mediated plant transformation and most of them focused on the binary vector system. A binary vector system comprises of a binary vector of which transferred DNA (T-DNA) resided on and a Ti plasmid to carry those essential virulence genes. In this study, we constructed a miniaturized helper Ti plasmid, designated as pYL102, with the aim to enhance the overall Agrobacterium-mediated transformation rate. The size of pYL102 was reduced to ~60% of the original plasmid pCAMBIA5105. Subsequently, pYL102 was coupled with the broad host range (BHR) bacterial expression vector, pYL101C, of which the key regulatory virulence gene, virG-N54D, was cloned in and expressed under the control of a strong constitutive PINTc promoter. To test the functionality of the constructed vector system, A. tumefaciens C58C1 carrying pYL102, pYL101C::virG-N54D and the transformation vector pGWB2::e35S-sfGFP was used to transform Nicotiana benthamiana leaves by agroinfiltration. Green fluorescence was observed in spots infiltrated with Agrobacterium carrying the test plasmids. The fluorescence intensity from the test agroinfiltrated leaves was significantly higher than those of the mock-infiltrated leaves (p<0.01), indicating the vector system can be used for plant transformation.
{"title":"Development of a miniaturized Ti-plasmid and helper plasmid system for Agrobacterium-mediated plant transformation","authors":"Yuh Leng Teo, Shu Ting Chang, W. K. Toh, C. Ho, P. Loh, H. Wong","doi":"10.35118/apjmbb.2022.030.3.03","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.03","url":null,"abstract":"Tumor-inducing (Ti) plasmid is the requisite for Agrobacterium-mediated plant transformation. Over decades, continuous efforts have been made to improve the efficiency of Agrobacterium-mediated plant transformation and most of them focused on the binary vector system. A binary vector system comprises of a binary vector of which transferred DNA (T-DNA) resided on and a Ti plasmid to carry those essential virulence genes. In this study, we constructed a miniaturized helper Ti plasmid, designated as pYL102, with the aim to enhance the overall Agrobacterium-mediated transformation rate. The size of pYL102 was reduced to ~60% of the original plasmid pCAMBIA5105. Subsequently, pYL102 was coupled with the broad host range (BHR) bacterial expression vector, pYL101C, of which the key regulatory virulence gene, virG-N54D, was cloned in and expressed under the control of a strong constitutive PINTc promoter. To test the functionality of the constructed vector system, A. tumefaciens C58C1 carrying pYL102, pYL101C::virG-N54D and the transformation vector pGWB2::e35S-sfGFP was used to transform Nicotiana benthamiana leaves by agroinfiltration. Green fluorescence was observed in spots infiltrated with Agrobacterium carrying the test plasmids. The fluorescence intensity from the test agroinfiltrated leaves was significantly higher than those of the mock-infiltrated leaves (p<0.01), indicating the vector system can be used for plant transformation.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78619159","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 : 2022-07-25DOI: 10.35118/apjmbb.2022.030.3.05
Shikta Rani Kar, Swapnila Choudhury, A. Chakraborty
The soybean is a valuable legume crop cultivated for its oil and protein which is used widely as food for humans and feed for livestock as well as in biofuel production. The genetic improvement of the soybean needs to be accelerated to boost its productivity and enhance its resilience to changing environments. In recent years, CRISPR/Cas9 has become a powerful and robust genome editing system for manipulating traits of various crop plants including soybean. This cutting-edge biotechnological tool has been extensively used as a means for improving crop quality and yields, disease-resistance, tolerance to adverse environmental conditions, and production of plant-based materials. This review presents a brief mechanism of the CRISPR/Cas9 system followed by its application in soybean improvement. It also highlights some prospects of using the CRISPR/Cas9 system in soybean research.
{"title":"CRISPR/Cas9 for soybean improvement: A review","authors":"Shikta Rani Kar, Swapnila Choudhury, A. Chakraborty","doi":"10.35118/apjmbb.2022.030.3.05","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.05","url":null,"abstract":"The soybean is a valuable legume crop cultivated for its oil and protein which is used widely as food for humans and feed for livestock as well as in biofuel production. The genetic improvement of the soybean needs to be accelerated to boost its productivity and enhance its resilience to changing environments. In recent years, CRISPR/Cas9 has become a powerful and robust genome editing system for manipulating traits of various crop plants including soybean. This cutting-edge biotechnological tool has been extensively used as a means for improving crop quality and yields, disease-resistance, tolerance to adverse environmental conditions, and production of plant-based materials. This review presents a brief mechanism of the CRISPR/Cas9 system followed by its application in soybean improvement. It also highlights some prospects of using the CRISPR/Cas9 system in soybean research.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89282905","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}
Each year, thousands of new cancer cases are reported all over the world. Among them, breast, colorectum, lung, nasopharynx and liver cancers are the top 5 cancer cases reported in 2018 in Malaysia (World Health Organization, 2020). In the same year, lung, colorectum, breast, liver and leukaemia cancer were the major death cancer cases in Malaysia. Radiotherapy, chemotherapy, and surgery are commonly used to treat cancer. However, they may cause some serious side effects apart from the common effects such as nausea and vomiting. Thus, more studies were done to search for alternative cancer treatments to reduce or eliminate these side effects. Vitamin E (tocopherol and tocotrienol) is one of the natural ingredients that were studied recently, and the compound was reported to show an anticancer properties. Tocopherol and tocotrienol were further divided into α-, β-, γ- and δ-tocopherol and tocotrienol but not all isomers were extensively studied. Most studies focusing on the examined isomers were in vitro investigations, with a few extending to in vivo studies. Only a few studies have progressed to clinical studies. This review aims to provide comprehensive information of previous studies on isoforms of vitamin E towards cancer studies.
{"title":"Vitamin E isomers and cancer research: A review","authors":"Atiqa Syazwani Ridzuan, Indah Mohd Amin, Khor Goot Heah, Rahayu Zulkapli","doi":"10.35118/apjmbb.2022.030.3.01","DOIUrl":"https://doi.org/10.35118/apjmbb.2022.030.3.01","url":null,"abstract":"Each year, thousands of new cancer cases are reported all over the world. Among them, breast, colorectum, lung, nasopharynx and liver cancers are the top 5 cancer cases reported in 2018 in Malaysia (World Health Organization, 2020). In the same year, lung, colorectum, breast, liver and leukaemia cancer were the major death cancer cases in Malaysia. Radiotherapy, chemotherapy, and surgery are commonly used to treat cancer. However, they may cause some serious side effects apart from the common effects such as nausea and vomiting. Thus, more studies were done to search for alternative cancer treatments to reduce or eliminate these side effects. Vitamin E (tocopherol and tocotrienol) is one of the natural ingredients that were studied recently, and the compound was reported to show an anticancer properties. Tocopherol and tocotrienol were further divided into α-, β-, γ- and δ-tocopherol and tocotrienol but not all isomers were extensively studied. Most studies focusing on the examined isomers were in vitro investigations, with a few extending to in vivo studies. Only a few studies have progressed to clinical studies. This review aims to provide comprehensive information of previous studies on isoforms of vitamin E towards cancer studies.","PeriodicalId":8566,"journal":{"name":"Asia-pacific Journal of Molecular Biology and Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81147895","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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