The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was det... more The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determined using restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects (NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an NTD-affected child.
Biotransformation is extensively used to create useful metabolites from various natural products ... more Biotransformation is extensively used to create useful metabolites from various natural products and as an alternative to chemical synthesis for the preparation of pharmacologically-active compounds. This study investigated the effects of asiaticoside and its biotransformed product on wound healing activities. Asiaticoside, the bioactive constituent of Centella asiatica has been reported to possess wound healing properties. Microbial transformation of asiaticoside using Aspergillus niger was carried out to produce an asiaticoside biotransformed product and the wound healing activities of asiaticoside and its biotransformed product were investigated. Their effects on transforming growth factor-beta 1 (TGFβ1) and tissue inhibitor of metalloproteinase 1 (TIMP1) gene expression were examined to understand the mode of action and on the cell proliferation and wound healing using human keratinocytes. Results of in vitro study showed that asiaticoside concentrations between 7.5 and 120 μg/m...
The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioact... more The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using microorganism. Asiaticoside was separated using an optimized HPLC method using Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50 V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at least two byproducts of the ancestor compound. In addition to that the preliminary screeing showed that the transformed product showed very high cytotoxicity agains HaCat Cells in comprison to the original compound.
Microbial transformation has been extensively used, to create new and useful metabolites.
Microbi... more Microbial transformation has been extensively used, to create new and useful metabolites. Microbial transformations can primarily be used in engineering and altering the effective constituents of the phytomedicine, increasing the natural products collection, and to draw the routes of drugs’ synthesis. Wound healing process normally starts with three subsequent actions that may be shared in period and area; these complicated stages include the inflammation phase, proliferation or tissue formation phase and remodeling phase. Asiaticoside was isolated from Centella asiatica and exposed to the microbial catalysis and then the metabolites were extracted. The effects of asiaticoside and its transformed products were investigated on human skin cells using human keratinocyte Hacat cell line as the in vitro model. In wound scratch assay on Hacat cells, asiaticoside and its microbial metabolome was shown to increase the migration rates of the cells. By tracking the area occupied by the cells and the number of cells attached, we could conclude that asiaticoside with its biocatalysed components increased the initial Hacat cell migration and adhesion. It was observed that the microbial metabolome demonstrated its effect with minimal concentration than its original compound. In cell proliferation assays, asiaticoside and the metabolome showed their efficacy in inducing the increase of cell behaviors. It can be assumed that the bioactivity on artificial skin, might suggest them to have pharmaceutical values in addition to the healing significance.
Ethyl p-methoxycinnamate (EPMC), a major constituent of the Kaempferia galanga rhizome,
was trans... more Ethyl p-methoxycinnamate (EPMC), a major constituent of the Kaempferia galanga rhizome, was transformed to ethyl p-hydroxycinnamate (EPHC) using Aspergillus niger. The EPHC metabolite was elucidated by NMR spectroscopic technique. Antimicrobial microbial study found that EPHC was active against all strains tested with a good minimum inhibitory concentration (MIC). It was active against both Staphylococcus aureus and Bacillus cereus at MIC 333 µg/ml while against Pseudomonas aeruginosa, Escherichia coli and Candida albicans at MIC 111 µg/ml. It was also shown that EPHC exhibited more growth inhibition potential than EPMC. Besides that, EPHC has shown the minimum bactericidal concentration (MBC) toward B. cereus, P. aeruginosa and E. coli at the concentration of 1000 µg/mL while EPMC did not show killing potential toward the tested microorganisms.
Studies were carried out on the extraction of the rhizome of Kaempferia galanga L. using steam di... more Studies were carried out on the extraction of the rhizome of Kaempferia galanga L. using steam distillation and supercritical fluid extraction (SFE). After purification, the major compound of the K. galanga L. extract, ethyl pmethoxycinnamate (EPMC) was identified by NMR and MS spectrometry. Biotransformation of EPMC using Aspergillus niger afforded ethyl p-hydroxycinnamate in 24% yield
This article revises the current state of microbial transformation use in natural products. It re... more This article revises the current state of microbial transformation use in natural products. It represents the results of the most recent reports. Due to the importance of this transformation, there is still a strong need to intensify the direction of microbial transformation of the natural product compounds, besides exploiting more microorganisms that might be the used biocatalysts in producing the novel materials for pharmaceutical purposes.
In recent years, artemisinin (1) has remained an effective drug to treat malaria. Numerous approa... more In recent years, artemisinin (1) has remained an effective drug to treat malaria. Numerous approaches have been adapted to increase the efficacy of (1) against antibiotic resistant malarial parasite. Microbial biotransformation of (1) has been used recently to produce promising derivatives of (1) on a large scale with low costs. During the last decade several biotransformation studies on (1), by using microorganisms, have been reported. This literature review focuses on the most recent microbial transformation studies on artemisinin and its derivatives.
The study was carried out to develop a rapid and accurate HPLC method to isolate the major
bioact... more The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using microorganism. Asiaticoside was separated using an optimized HPLC method using Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50 V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at least two byproducts of the ancestor compound. In addition to that the preliminary screeing showed that the transformed product showed very high cytotoxicity agains HaCat Cells in comprison to the original compound.
Lignin is an important constitute of the wood and thus the organism like fungi
and insects growin... more Lignin is an important constitute of the wood and thus the organism like fungi and insects growing on wood produce some extracellular lignolytic enzymes. In fungi these enzymes include different oxidases and peroxidases. Lignin peroxidases, manganese peroxidases, versatile peroxidases and laccases have been characterised from fungi and their roles in wood decay has been established. Here we report ABTS-oxidases from an unidentified fungus isolated from tree bark sample from Kuantan, Malaysia. After the liquid media cultivation, the crude extracellular protein content was extracted and analysed by polyacrylamide gel electrophoresis. SDS-PAGE, native PAGE and zymography were carried out. Azo dyes, brilliant green, trypan blue, direct red 80 and eriochrome black T were used for decolourisation activity of the isolated fungus on solid medium. The results suggest that the extracellular proteins contain laccase enzyme activity. Out of the four azo dyes that were used for dye decolourisation assay, at least three were decolourised to different extents by the fungus.
The main aim of the current study is to isolate a fungi with laccase activities to be used for tr... more The main aim of the current study is to isolate a fungi with laccase activities to be used for transformation of phytochemicals isolated from the same environment as an approach to produce novel pharmaceutical products. In our study, laccase producing fungi was isolated from samples taken from a tropical trees barks which grown in the International Islamic University Malaysia, Kuantan campus using different chromogenic substrates on the agar plates such as ABTS, Tannic Acid, Azure B and Guaiacol. It was found the isolated fungi has excellent ability to produce laccase. The conclusion of this study confirm here that using ABTS is very useful to isolate laccase producing fungus. The microscopic visualization showed the characteristics of white-rot fungi. Further molecular identification still ongoing to check the complete identity of our fungi.
Centella asiatica or “gotu kola” has been used since long time ago as an ethno-pharmacological pl... more Centella asiatica or “gotu kola” has been used since long time ago as an ethno-pharmacological plant and supposed to be a potent medicinal plant for its various pharmacological effects favorable for human health. Many studies described the noteworthy protective effect of C. asiatica against numerous diseases. Biological activities of C. asiatica have been linked to the most major compounds in it. This state paper is a part of our ongoing research on C. asiatica, its activity, isolation of novel metabolites, and applying the biotechnological methods to improve this plant and its phytochemical activities.
Neural tube defects (NTDs) are the second most common severely disabling human congenital defects... more Neural tube defects (NTDs) are the second most common severely disabling human congenital defects. Worldwide, NTDs incidence is approximately one per 1000 live births and varies between 0.78 and 12 per 1000 births in general populations. NTDs causation involves multiple genes, nutritional and environmental factors. Research in the next stage should include bigger populations and bigger studies that would be suitable to detect significant links and conclusions with relatively minor risk factors; analysis of multiple candidate genes to detect potential gene-gene interactions; detailed analysis of patient samples stratified by phenotype.
BACKGROUND:
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105... more BACKGROUND:
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105.24 kb and is made up of 33 exons. Methionine synthase is a cytoplasmic enzyme that requires methylcobalamin for activity and catalyzes the remethylation of homocysteine to methionine. In this reaction, the methyl group of 5-methyltetrahydrofolate is transferred to the enzyme bond cob(I) alamin to generate methylcobalamin, followed by the transfer of the methyl group to homocysteine to reform methionine.
MATERIALS AND METHODS:
The frequencies of the polymorphisms of MTR 2756A>G and MTR 2758C>G have been determined in this study in a sample of 491 individuals collected from all regions of Jordan and representing the Jordanian population. The different alleles and genotypes at the two polymorphic sites were identified using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.
RESULTS:
Showed that the percentages of the polymorphic alleles at the MTR 2756 position in the north, middle and south regions were 90.38, 92.65 and 83.69%, respectively, for the MTR 2756A allele, and were 9.61, 7.34 and 16.30%, respectively, for the MTR 2756G allele, with overall percentages in the whole Jordanian population of 90.73 and 9.27% for the MTR 2756A and MTR 2756G alleles, respectively. The percentages of the genotype MTR 2756AA were 82.90% in the northern region, 86.72% in the middle region and 71.73% in the southern region, and an overall percentage of MTR 2756AA in the whole Jordanian population was 83.50%. The frequencies of MTR 2756AG genotype in the northern, middle and southern regions were 14.95, 11.84 and 23.91%, respectively, with an overall percentage of 14.46% in the whole Jordanian population. The percentages of the genotype MTR 2756GG in the northern, middle and southern regions were 2.13, 1.42 and 4.34%, respectively, with an overall percentage of 2.04% in the whole Jordanian population. Only the wild type allele (C) of the MTR 2758C>G polymorphism was detected in this study. In addition, the association of MTR 2756A>G and MTR 2758C>G polymorphisms with the development of neural tube defects (NTDs) was examined using 17 cases of mothers from the northern part of Jordan, who gave birth to NTD affected children during the period of this study. Results showed no association between these two examined polymorphisms and the increase in maternal risk for giving birth to NTD children.
CONCLUSION:
results of this study recommend that examination should be done on larger populations to arrive at better conclusions. Also, more studies on gene–gene interaction should be done to examine the associations with NTDs.
The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determ... more The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determined using restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects (NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an NTD-affected child
The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was det... more The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determined using restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects (NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an NTD-affected child.
Biotransformation is extensively used to create useful metabolites from various natural products ... more Biotransformation is extensively used to create useful metabolites from various natural products and as an alternative to chemical synthesis for the preparation of pharmacologically-active compounds. This study investigated the effects of asiaticoside and its biotransformed product on wound healing activities. Asiaticoside, the bioactive constituent of Centella asiatica has been reported to possess wound healing properties. Microbial transformation of asiaticoside using Aspergillus niger was carried out to produce an asiaticoside biotransformed product and the wound healing activities of asiaticoside and its biotransformed product were investigated. Their effects on transforming growth factor-beta 1 (TGFβ1) and tissue inhibitor of metalloproteinase 1 (TIMP1) gene expression were examined to understand the mode of action and on the cell proliferation and wound healing using human keratinocytes. Results of in vitro study showed that asiaticoside concentrations between 7.5 and 120 μg/m...
The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioact... more The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using microorganism. Asiaticoside was separated using an optimized HPLC method using Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50 V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at least two byproducts of the ancestor compound. In addition to that the preliminary screeing showed that the transformed product showed very high cytotoxicity agains HaCat Cells in comprison to the original compound.
Microbial transformation has been extensively used, to create new and useful metabolites.
Microbi... more Microbial transformation has been extensively used, to create new and useful metabolites. Microbial transformations can primarily be used in engineering and altering the effective constituents of the phytomedicine, increasing the natural products collection, and to draw the routes of drugs’ synthesis. Wound healing process normally starts with three subsequent actions that may be shared in period and area; these complicated stages include the inflammation phase, proliferation or tissue formation phase and remodeling phase. Asiaticoside was isolated from Centella asiatica and exposed to the microbial catalysis and then the metabolites were extracted. The effects of asiaticoside and its transformed products were investigated on human skin cells using human keratinocyte Hacat cell line as the in vitro model. In wound scratch assay on Hacat cells, asiaticoside and its microbial metabolome was shown to increase the migration rates of the cells. By tracking the area occupied by the cells and the number of cells attached, we could conclude that asiaticoside with its biocatalysed components increased the initial Hacat cell migration and adhesion. It was observed that the microbial metabolome demonstrated its effect with minimal concentration than its original compound. In cell proliferation assays, asiaticoside and the metabolome showed their efficacy in inducing the increase of cell behaviors. It can be assumed that the bioactivity on artificial skin, might suggest them to have pharmaceutical values in addition to the healing significance.
Ethyl p-methoxycinnamate (EPMC), a major constituent of the Kaempferia galanga rhizome,
was trans... more Ethyl p-methoxycinnamate (EPMC), a major constituent of the Kaempferia galanga rhizome, was transformed to ethyl p-hydroxycinnamate (EPHC) using Aspergillus niger. The EPHC metabolite was elucidated by NMR spectroscopic technique. Antimicrobial microbial study found that EPHC was active against all strains tested with a good minimum inhibitory concentration (MIC). It was active against both Staphylococcus aureus and Bacillus cereus at MIC 333 µg/ml while against Pseudomonas aeruginosa, Escherichia coli and Candida albicans at MIC 111 µg/ml. It was also shown that EPHC exhibited more growth inhibition potential than EPMC. Besides that, EPHC has shown the minimum bactericidal concentration (MBC) toward B. cereus, P. aeruginosa and E. coli at the concentration of 1000 µg/mL while EPMC did not show killing potential toward the tested microorganisms.
Studies were carried out on the extraction of the rhizome of Kaempferia galanga L. using steam di... more Studies were carried out on the extraction of the rhizome of Kaempferia galanga L. using steam distillation and supercritical fluid extraction (SFE). After purification, the major compound of the K. galanga L. extract, ethyl pmethoxycinnamate (EPMC) was identified by NMR and MS spectrometry. Biotransformation of EPMC using Aspergillus niger afforded ethyl p-hydroxycinnamate in 24% yield
This article revises the current state of microbial transformation use in natural products. It re... more This article revises the current state of microbial transformation use in natural products. It represents the results of the most recent reports. Due to the importance of this transformation, there is still a strong need to intensify the direction of microbial transformation of the natural product compounds, besides exploiting more microorganisms that might be the used biocatalysts in producing the novel materials for pharmaceutical purposes.
In recent years, artemisinin (1) has remained an effective drug to treat malaria. Numerous approa... more In recent years, artemisinin (1) has remained an effective drug to treat malaria. Numerous approaches have been adapted to increase the efficacy of (1) against antibiotic resistant malarial parasite. Microbial biotransformation of (1) has been used recently to produce promising derivatives of (1) on a large scale with low costs. During the last decade several biotransformation studies on (1), by using microorganisms, have been reported. This literature review focuses on the most recent microbial transformation studies on artemisinin and its derivatives.
The study was carried out to develop a rapid and accurate HPLC method to isolate the major
bioact... more The study was carried out to develop a rapid and accurate HPLC method to isolate the major bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using microorganism. Asiaticoside was separated using an optimized HPLC method using Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50 V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at least two byproducts of the ancestor compound. In addition to that the preliminary screeing showed that the transformed product showed very high cytotoxicity agains HaCat Cells in comprison to the original compound.
Lignin is an important constitute of the wood and thus the organism like fungi
and insects growin... more Lignin is an important constitute of the wood and thus the organism like fungi and insects growing on wood produce some extracellular lignolytic enzymes. In fungi these enzymes include different oxidases and peroxidases. Lignin peroxidases, manganese peroxidases, versatile peroxidases and laccases have been characterised from fungi and their roles in wood decay has been established. Here we report ABTS-oxidases from an unidentified fungus isolated from tree bark sample from Kuantan, Malaysia. After the liquid media cultivation, the crude extracellular protein content was extracted and analysed by polyacrylamide gel electrophoresis. SDS-PAGE, native PAGE and zymography were carried out. Azo dyes, brilliant green, trypan blue, direct red 80 and eriochrome black T were used for decolourisation activity of the isolated fungus on solid medium. The results suggest that the extracellular proteins contain laccase enzyme activity. Out of the four azo dyes that were used for dye decolourisation assay, at least three were decolourised to different extents by the fungus.
The main aim of the current study is to isolate a fungi with laccase activities to be used for tr... more The main aim of the current study is to isolate a fungi with laccase activities to be used for transformation of phytochemicals isolated from the same environment as an approach to produce novel pharmaceutical products. In our study, laccase producing fungi was isolated from samples taken from a tropical trees barks which grown in the International Islamic University Malaysia, Kuantan campus using different chromogenic substrates on the agar plates such as ABTS, Tannic Acid, Azure B and Guaiacol. It was found the isolated fungi has excellent ability to produce laccase. The conclusion of this study confirm here that using ABTS is very useful to isolate laccase producing fungus. The microscopic visualization showed the characteristics of white-rot fungi. Further molecular identification still ongoing to check the complete identity of our fungi.
Centella asiatica or “gotu kola” has been used since long time ago as an ethno-pharmacological pl... more Centella asiatica or “gotu kola” has been used since long time ago as an ethno-pharmacological plant and supposed to be a potent medicinal plant for its various pharmacological effects favorable for human health. Many studies described the noteworthy protective effect of C. asiatica against numerous diseases. Biological activities of C. asiatica have been linked to the most major compounds in it. This state paper is a part of our ongoing research on C. asiatica, its activity, isolation of novel metabolites, and applying the biotechnological methods to improve this plant and its phytochemical activities.
Neural tube defects (NTDs) are the second most common severely disabling human congenital defects... more Neural tube defects (NTDs) are the second most common severely disabling human congenital defects. Worldwide, NTDs incidence is approximately one per 1000 live births and varies between 0.78 and 12 per 1000 births in general populations. NTDs causation involves multiple genes, nutritional and environmental factors. Research in the next stage should include bigger populations and bigger studies that would be suitable to detect significant links and conclusions with relatively minor risk factors; analysis of multiple candidate genes to detect potential gene-gene interactions; detailed analysis of patient samples stratified by phenotype.
BACKGROUND:
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105... more BACKGROUND:
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105.24 kb and is made up of 33 exons. Methionine synthase is a cytoplasmic enzyme that requires methylcobalamin for activity and catalyzes the remethylation of homocysteine to methionine. In this reaction, the methyl group of 5-methyltetrahydrofolate is transferred to the enzyme bond cob(I) alamin to generate methylcobalamin, followed by the transfer of the methyl group to homocysteine to reform methionine.
MATERIALS AND METHODS:
The frequencies of the polymorphisms of MTR 2756A>G and MTR 2758C>G have been determined in this study in a sample of 491 individuals collected from all regions of Jordan and representing the Jordanian population. The different alleles and genotypes at the two polymorphic sites were identified using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.
RESULTS:
Showed that the percentages of the polymorphic alleles at the MTR 2756 position in the north, middle and south regions were 90.38, 92.65 and 83.69%, respectively, for the MTR 2756A allele, and were 9.61, 7.34 and 16.30%, respectively, for the MTR 2756G allele, with overall percentages in the whole Jordanian population of 90.73 and 9.27% for the MTR 2756A and MTR 2756G alleles, respectively. The percentages of the genotype MTR 2756AA were 82.90% in the northern region, 86.72% in the middle region and 71.73% in the southern region, and an overall percentage of MTR 2756AA in the whole Jordanian population was 83.50%. The frequencies of MTR 2756AG genotype in the northern, middle and southern regions were 14.95, 11.84 and 23.91%, respectively, with an overall percentage of 14.46% in the whole Jordanian population. The percentages of the genotype MTR 2756GG in the northern, middle and southern regions were 2.13, 1.42 and 4.34%, respectively, with an overall percentage of 2.04% in the whole Jordanian population. Only the wild type allele (C) of the MTR 2758C>G polymorphism was detected in this study. In addition, the association of MTR 2756A>G and MTR 2758C>G polymorphisms with the development of neural tube defects (NTDs) was examined using 17 cases of mothers from the northern part of Jordan, who gave birth to NTD affected children during the period of this study. Results showed no association between these two examined polymorphisms and the increase in maternal risk for giving birth to NTD children.
CONCLUSION:
results of this study recommend that examination should be done on larger populations to arrive at better conclusions. Also, more studies on gene–gene interaction should be done to examine the associations with NTDs.
The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determ... more The frequency of methylenetetrahydrofolate dehydrogenase (MTHFD1) 1958G>A polymorphism was determined using restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects (NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an NTD-affected child
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Papers by Helmi Y Alfarra
Microbial transformations can primarily be used in engineering and altering the effective
constituents of the phytomedicine, increasing the natural products collection, and to draw the
routes of drugs’ synthesis. Wound healing process normally starts with three subsequent
actions that may be shared in period and area; these complicated stages include the
inflammation phase, proliferation or tissue formation phase and remodeling phase.
Asiaticoside was isolated from Centella asiatica and exposed to the microbial catalysis and
then the metabolites were extracted. The effects of asiaticoside and its transformed products
were investigated on human skin cells using human keratinocyte Hacat cell line as the in vitro
model. In wound scratch assay on Hacat cells, asiaticoside and its microbial metabolome was
shown to increase the migration rates of the cells. By tracking the area occupied by the cells
and the number of cells attached, we could conclude that asiaticoside with its biocatalysed
components increased the initial Hacat cell migration and adhesion. It was observed that the
microbial metabolome demonstrated its effect with minimal concentration than its original
compound. In cell proliferation assays, asiaticoside and the metabolome showed their efficacy
in inducing the increase of cell behaviors. It can be assumed that the bioactivity on artificial
skin, might suggest them to have pharmaceutical values in addition to the healing
significance.
was transformed to ethyl p-hydroxycinnamate (EPHC) using Aspergillus niger. The EPHC metabolite
was elucidated by NMR spectroscopic technique. Antimicrobial microbial study found that EPHC
was active against all strains tested with a good minimum inhibitory concentration (MIC). It was
active against both Staphylococcus aureus and Bacillus cereus at MIC 333 µg/ml while against
Pseudomonas aeruginosa, Escherichia coli and Candida albicans at MIC 111 µg/ml. It was also
shown that EPHC exhibited more growth inhibition potential than EPMC. Besides that, EPHC has
shown the minimum bactericidal concentration (MBC) toward B. cereus, P. aeruginosa and E. coli
at the concentration of 1000 µg/mL while EPMC did not show killing potential toward the tested
microorganisms.
supercritical fluid extraction (SFE). After purification, the major compound of the K. galanga L. extract, ethyl pmethoxycinnamate
(EPMC) was identified by NMR and MS spectrometry. Biotransformation of EPMC using
Aspergillus niger afforded ethyl p-hydroxycinnamate in 24% yield
most recent reports. Due to the importance of this transformation, there is still a strong need to intensify the direction of
microbial transformation of the natural product compounds, besides exploiting more microorganisms that might be the
used biocatalysts in producing the novel materials for pharmaceutical purposes.
increase the efficacy of (1) against antibiotic resistant malarial parasite. Microbial biotransformation of (1) has been used
recently to produce promising derivatives of (1) on a large scale with low costs. During the last decade several
biotransformation studies on (1), by using microorganisms, have been reported. This literature review focuses on the most
recent microbial transformation studies on artemisinin and its derivatives.
bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using
microorganism. Asiaticoside was separated using an optimized HPLC method using
Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50
V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate
with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the
prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at
least two byproducts of the ancestor compound. In addition to that the preliminary screeing
showed that the transformed product showed very high cytotoxicity agains HaCat Cells in
comprison to the original compound.
and insects growing on wood produce some extracellular lignolytic enzymes. In fungi
these enzymes include different oxidases and peroxidases. Lignin peroxidases, manganese
peroxidases, versatile peroxidases and laccases have been characterised from fungi and
their roles in wood decay has been established. Here we report ABTS-oxidases from an
unidentified fungus isolated from tree bark sample from Kuantan, Malaysia. After the
liquid media cultivation, the crude extracellular protein content was extracted and
analysed by polyacrylamide gel electrophoresis. SDS-PAGE, native PAGE and zymography
were carried out. Azo dyes, brilliant green, trypan blue, direct red 80 and eriochrome
black T were used for decolourisation activity of the isolated fungus on solid medium.
The results suggest that the extracellular proteins contain laccase enzyme activity. Out of
the four azo dyes that were used for dye decolourisation assay, at least three were
decolourised to different extents by the fungus.
isolated from the same environment as an approach to produce novel pharmaceutical products. In our study, laccase
producing fungi was isolated from samples taken from a tropical trees barks which grown in the International Islamic
University Malaysia, Kuantan campus using different chromogenic substrates on the agar plates such as ABTS, Tannic Acid,
Azure B and Guaiacol. It was found the isolated fungi has excellent ability to produce laccase. The conclusion of this study
confirm here that using ABTS is very useful to isolate laccase producing fungus. The microscopic visualization showed the
characteristics of white-rot fungi. Further molecular identification still ongoing to check the complete identity of our fungi.
supposed to be a potent medicinal plant for its various pharmacological effects favorable for human health. Many
studies described the noteworthy protective effect of C. asiatica against numerous diseases. Biological activities
of C. asiatica have been linked to the most major compounds in it. This state paper is a part of our ongoing
research on C. asiatica, its activity, isolation of novel metabolites, and applying the biotechnological methods to
improve this plant and its phytochemical activities.
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105.24 kb and is made up of 33 exons. Methionine synthase is a cytoplasmic enzyme that requires methylcobalamin for activity and catalyzes the remethylation of homocysteine to methionine. In this reaction, the methyl group of 5-methyltetrahydrofolate is transferred to the enzyme bond cob(I) alamin to generate methylcobalamin, followed by the transfer of the methyl group to homocysteine to reform methionine.
MATERIALS AND METHODS:
The frequencies of the polymorphisms of MTR 2756A>G and MTR 2758C>G have been determined in this study in a sample of 491 individuals collected from all regions of Jordan and representing the Jordanian population. The different alleles and genotypes at the two polymorphic sites were identified using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.
RESULTS:
Showed that the percentages of the polymorphic alleles at the MTR 2756 position in the north, middle and south regions were 90.38, 92.65 and 83.69%, respectively, for the MTR 2756A allele, and were 9.61, 7.34 and 16.30%, respectively, for the MTR 2756G allele, with overall percentages in the whole Jordanian population of 90.73 and 9.27% for the MTR 2756A and MTR 2756G alleles, respectively. The percentages of the genotype MTR 2756AA were 82.90% in the northern region, 86.72% in the middle region and 71.73% in the southern region, and an overall percentage of MTR 2756AA in the whole Jordanian population was 83.50%. The frequencies of MTR 2756AG genotype in the northern, middle and southern regions were 14.95, 11.84 and 23.91%, respectively, with an overall percentage of 14.46% in the whole Jordanian population. The percentages of the genotype MTR 2756GG in the northern, middle and southern regions were 2.13, 1.42 and 4.34%, respectively, with an overall percentage of 2.04% in the whole Jordanian population. Only the wild type allele (C) of the MTR 2758C>G polymorphism was detected in this study. In addition, the association of MTR 2756A>G and MTR 2758C>G polymorphisms with the development of neural tube defects (NTDs) was examined using 17 cases of mothers from the northern part of Jordan, who gave birth to NTD affected children during the period of this study. Results showed no association between these two examined polymorphisms and the increase in maternal risk for giving birth to NTD children.
CONCLUSION:
results of this study recommend that examination should be done on larger populations to arrive at better conclusions. Also, more studies on gene–gene interaction should be done to examine the associations with NTDs.
restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different
regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the
G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes
respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the
different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects
(NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the
period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an
NTD-affected child
Microbial transformations can primarily be used in engineering and altering the effective
constituents of the phytomedicine, increasing the natural products collection, and to draw the
routes of drugs’ synthesis. Wound healing process normally starts with three subsequent
actions that may be shared in period and area; these complicated stages include the
inflammation phase, proliferation or tissue formation phase and remodeling phase.
Asiaticoside was isolated from Centella asiatica and exposed to the microbial catalysis and
then the metabolites were extracted. The effects of asiaticoside and its transformed products
were investigated on human skin cells using human keratinocyte Hacat cell line as the in vitro
model. In wound scratch assay on Hacat cells, asiaticoside and its microbial metabolome was
shown to increase the migration rates of the cells. By tracking the area occupied by the cells
and the number of cells attached, we could conclude that asiaticoside with its biocatalysed
components increased the initial Hacat cell migration and adhesion. It was observed that the
microbial metabolome demonstrated its effect with minimal concentration than its original
compound. In cell proliferation assays, asiaticoside and the metabolome showed their efficacy
in inducing the increase of cell behaviors. It can be assumed that the bioactivity on artificial
skin, might suggest them to have pharmaceutical values in addition to the healing
significance.
was transformed to ethyl p-hydroxycinnamate (EPHC) using Aspergillus niger. The EPHC metabolite
was elucidated by NMR spectroscopic technique. Antimicrobial microbial study found that EPHC
was active against all strains tested with a good minimum inhibitory concentration (MIC). It was
active against both Staphylococcus aureus and Bacillus cereus at MIC 333 µg/ml while against
Pseudomonas aeruginosa, Escherichia coli and Candida albicans at MIC 111 µg/ml. It was also
shown that EPHC exhibited more growth inhibition potential than EPMC. Besides that, EPHC has
shown the minimum bactericidal concentration (MBC) toward B. cereus, P. aeruginosa and E. coli
at the concentration of 1000 µg/mL while EPMC did not show killing potential toward the tested
microorganisms.
supercritical fluid extraction (SFE). After purification, the major compound of the K. galanga L. extract, ethyl pmethoxycinnamate
(EPMC) was identified by NMR and MS spectrometry. Biotransformation of EPMC using
Aspergillus niger afforded ethyl p-hydroxycinnamate in 24% yield
most recent reports. Due to the importance of this transformation, there is still a strong need to intensify the direction of
microbial transformation of the natural product compounds, besides exploiting more microorganisms that might be the
used biocatalysts in producing the novel materials for pharmaceutical purposes.
increase the efficacy of (1) against antibiotic resistant malarial parasite. Microbial biotransformation of (1) has been used
recently to produce promising derivatives of (1) on a large scale with low costs. During the last decade several
biotransformation studies on (1), by using microorganisms, have been reported. This literature review focuses on the most
recent microbial transformation studies on artemisinin and its derivatives.
bioactive phytochemical asiaticoside in Centella asiatica and to transform this metabolite using
microorganism. Asiaticoside was separated using an optimized HPLC method using
Phenomenex, Luna, 5 μm (C18) (150 mm x 4.6 mm) eluted with Water PH3: Acetonitrile (50:50
V/V). The reported method is easy, fast and accurate, and produced highly pure, resolved isolate
with high concentration. Asiaticoside was biologically transformed by Aspergillus niger, and the
prescreening results on TLC and 1D and 2D IR showed that the transformed produced has at
least two byproducts of the ancestor compound. In addition to that the preliminary screeing
showed that the transformed product showed very high cytotoxicity agains HaCat Cells in
comprison to the original compound.
and insects growing on wood produce some extracellular lignolytic enzymes. In fungi
these enzymes include different oxidases and peroxidases. Lignin peroxidases, manganese
peroxidases, versatile peroxidases and laccases have been characterised from fungi and
their roles in wood decay has been established. Here we report ABTS-oxidases from an
unidentified fungus isolated from tree bark sample from Kuantan, Malaysia. After the
liquid media cultivation, the crude extracellular protein content was extracted and
analysed by polyacrylamide gel electrophoresis. SDS-PAGE, native PAGE and zymography
were carried out. Azo dyes, brilliant green, trypan blue, direct red 80 and eriochrome
black T were used for decolourisation activity of the isolated fungus on solid medium.
The results suggest that the extracellular proteins contain laccase enzyme activity. Out of
the four azo dyes that were used for dye decolourisation assay, at least three were
decolourised to different extents by the fungus.
isolated from the same environment as an approach to produce novel pharmaceutical products. In our study, laccase
producing fungi was isolated from samples taken from a tropical trees barks which grown in the International Islamic
University Malaysia, Kuantan campus using different chromogenic substrates on the agar plates such as ABTS, Tannic Acid,
Azure B and Guaiacol. It was found the isolated fungi has excellent ability to produce laccase. The conclusion of this study
confirm here that using ABTS is very useful to isolate laccase producing fungus. The microscopic visualization showed the
characteristics of white-rot fungi. Further molecular identification still ongoing to check the complete identity of our fungi.
supposed to be a potent medicinal plant for its various pharmacological effects favorable for human health. Many
studies described the noteworthy protective effect of C. asiatica against numerous diseases. Biological activities
of C. asiatica have been linked to the most major compounds in it. This state paper is a part of our ongoing
research on C. asiatica, its activity, isolation of novel metabolites, and applying the biotechnological methods to
improve this plant and its phytochemical activities.
The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105.24 kb and is made up of 33 exons. Methionine synthase is a cytoplasmic enzyme that requires methylcobalamin for activity and catalyzes the remethylation of homocysteine to methionine. In this reaction, the methyl group of 5-methyltetrahydrofolate is transferred to the enzyme bond cob(I) alamin to generate methylcobalamin, followed by the transfer of the methyl group to homocysteine to reform methionine.
MATERIALS AND METHODS:
The frequencies of the polymorphisms of MTR 2756A>G and MTR 2758C>G have been determined in this study in a sample of 491 individuals collected from all regions of Jordan and representing the Jordanian population. The different alleles and genotypes at the two polymorphic sites were identified using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.
RESULTS:
Showed that the percentages of the polymorphic alleles at the MTR 2756 position in the north, middle and south regions were 90.38, 92.65 and 83.69%, respectively, for the MTR 2756A allele, and were 9.61, 7.34 and 16.30%, respectively, for the MTR 2756G allele, with overall percentages in the whole Jordanian population of 90.73 and 9.27% for the MTR 2756A and MTR 2756G alleles, respectively. The percentages of the genotype MTR 2756AA were 82.90% in the northern region, 86.72% in the middle region and 71.73% in the southern region, and an overall percentage of MTR 2756AA in the whole Jordanian population was 83.50%. The frequencies of MTR 2756AG genotype in the northern, middle and southern regions were 14.95, 11.84 and 23.91%, respectively, with an overall percentage of 14.46% in the whole Jordanian population. The percentages of the genotype MTR 2756GG in the northern, middle and southern regions were 2.13, 1.42 and 4.34%, respectively, with an overall percentage of 2.04% in the whole Jordanian population. Only the wild type allele (C) of the MTR 2758C>G polymorphism was detected in this study. In addition, the association of MTR 2756A>G and MTR 2758C>G polymorphisms with the development of neural tube defects (NTDs) was examined using 17 cases of mothers from the northern part of Jordan, who gave birth to NTD affected children during the period of this study. Results showed no association between these two examined polymorphisms and the increase in maternal risk for giving birth to NTD children.
CONCLUSION:
results of this study recommend that examination should be done on larger populations to arrive at better conclusions. Also, more studies on gene–gene interaction should be done to examine the associations with NTDs.
restriction fragment length polymorphism polymerase chain reaction analysis in a sample of 491 individuals from different
regions of Jordan. The distribution of polymorphic alleles of MTHFD1 1958G>A in the Jordanian population were 65.2% for the
G allele and 34.9% for the A allele. The genotype distributions were 48.7%, 32.8% and 18.5% for the GG, GA and AA genotypes
respectively. Statistical analysis showed no significant correlation between the rates of alleles and genotypes across the
different regions of Jordan. The association of MTHFD1 1958G>A polymorphism with the development of neural tube defects
(NTDs) was examined for 17 mothers from the northern part of Jordan who gave birth to an NTD-affected child during the
period of this study. The results showed no association between these 3 examined polymorphisms and maternal risk for an
NTD-affected child