Evaluating inhibitory potential targeting cholesteryl ester transfer protein (CETP) by hydroxycitric acid (HCA) found in garcinia species through kinetic and in-silico technique

Cardiovascular disease has emerged in developing countries and becoming the leading cause of death recorded. Many scientific studies have been conducted in order to understand the specific mechanism on how atherosclerosis develop, searching for the real culprit that responsible in the progression of the disease and suggesting the possible prevention to overcome this problem. This piece of work examined and revealed the mechanism of action on how secondary metabolites that has been isolated from Malaysian local plants which have the properties to impede the action of cholesteryl ester transfer protein (CETP) in order to prevent the atherosclerosis. Preliminary results of the crude plant extracts from the initial screening showed positive results. A similar trend of inhibition can be obtained for twigs and leaves extracts of Garcinia atroviridis and Garcinia parvifolia. Ethanol extracts of fruit parts of Garcinia atroviridis give IC50 of 19.28 ± 0.021 mg/ml which shows the highest inhibitory compared to the other extracts of other plant parts. The remarkable results that are obtain from fruit rinds of Garcinia atroviridis do give some hints that the secondary metabolites that are present might have the ability to inhibit CETP. Based on literature review, it is postulated hydroxycitric acid (HCA) might be responsible for inhibiting CETP activity and HCA has been selected for further studies. Kinetic studies have been employed in this piece of work in order to see the types of inhibition that HCA possess against CETP. The kinetic study has revealed that HCA is a noncompetitive inhibitor because of the Km (-0.12) that is unchanged for every substrate and the Vmax is increased when the concentration of the inhibitor increase. Further in-silico works such as molecular docking and molecular dynamic has been implemented as well in order to see the interaction and mechanism of action between HCA and CETP. The molecular docking work has revealed that HCA binds to the same side as torcetrapib does and the RMSD obtained was 2.703Å. Molecular dynamics has been employed as well in order to see the extensive structural and functional analysis and also to evaluate the strengthness of the complex between HCA and CETP. The complex were found to be stable due to the existence of the hydrogen bonding to SER230 and the overall RMSD reading are between the range of 0.8Å, 2.4Å and 3.2Å. Overall, this work are pioneering and pave the way for further studies in establishing a new chemical template form of natural products for CETP research with an objective to extend the scope of work into in-vivo studies and x-ray crystallography in order to enable us to understand the mechanism of action in protein level. The in-silico studies in this work provides a preliminary understanding on the structural basis of CETP structure and its active sites which could accommodate the exact template of chemical molecule. With this new understanding, an inhibitor drug which are effective with lesser side effect, targeting atherosclerosis could be developed

Evaluating inhibitory potential targeting cholesteryl ester transfer protein (CETP) by hydroxycitric acid (HCA) found in garcinia species through kinetic and in-silico technique

Cardiovascular disease has emerged in developing countries and becoming the leading cause of death recorded. Many scientific studies have been conducted in order to understand the specific mechanism on how atherosclerosis develop, searching for the real culprit that responsible in the progression of the disease and suggesting the possible prevention to overcome this problem. This piece of work examined and revealed the mechanism of action on how secondary metabolites that has been isolated from Malaysian local plants which have the properties to impede the action of cholesteryl ester transfer protein (CETP) in order to prevent the atherosclerosis. Preliminary results of the crude plant extracts from the initial screening showed positive results. A similar trend of inhibition can be obtained for twigs and leaves extracts of Garcinia atroviridis and Garcinia parvifolia. Ethanol extracts of fruit parts of Garcinia atroviridis give IC50 of 19.28 ± 0.021 mg/ml which shows the highest inhibitory compared to the other extracts of other plant parts. The remarkable results that are obtain from fruit rinds of Garcinia atroviridis do give some hints that the secondary metabolites that are present might have the ability to inhibit CETP. Based on literature review, it is postulated hydroxycitric acid (HCA) might be responsible for inhibiting CETP activity and HCA has been selected for further studies. Kinetic studies have been employed in this piece of work in order to see the types of inhibition that HCA possess against CETP. The kinetic study has revealed that HCA is a noncompetitive inhibitor because of the Km (-0.12) that is unchanged for every substrate and the Vmax is increased when the concentration of the inhibitor increase. Further in-silico works such as molecular docking and molecular dynamic has been implemented as well in order to see the interaction and mechanism of action between HCA and CETP. The molecular docking work has revealed that HCA binds to the same side as torcetrapib does and the RMSD obtained was 2.703Å. Molecular dynamics has been employed as well in order to see the extensive structural and functional analysis and also to evaluate the strengthness of the complex between HCA and CETP. The complex were found to be stable due to the existence of the hydrogen bonding to SER230 and the overall RMSD reading are between the range of 0.8Å, 2.4Å and 3.2Å. Overall, this work are pioneering and pave the way for further studies in establishing a new chemical template form of natural products for CETP research with an objective to extend the scope of work into in-vivo studies and x-ray crystallography in order to enable us to understand the mechanism of action in protein level. The in-silico studies in this work provides a preliminary understanding on the structural basis of CETP structure and its active sites which could accommodate the exact template of chemical molecule. With this new understanding, an inhibitor drug which are effective with lesser side effect, targeting atherosclerosis could be developed

In-silico approach towards identification of inhibitor for cholesteryl ester transfer protein (CETP): A novel strategy

Atherosclerosis is a multifactorial disease which caused by a long term process of an accumulation of lipids combines with an inflammatory response which forms plaques or atheroma. The current focus in preventing atherosclerosis is by raising the HDL-C level. One novel approach is by inhibition of Cholesterol Ester Transfer Protein (CETP) function. Aim: The aim of this study is to see whether HCA does bind to the same active site in the CETP tunnel and to see the stability of the ligand-protein binding. Method: Docking studies are carried out by using X-ray crystallography structure (PDB ID: 20BD and 4EWS) by using Glides software from Schrodinger Inc and validated results using GOLD. The molecular dynamic simulation are being carried out by Desmond (Schrodinger inc) by using the ligand-protein complexes from the docking results. Results: Based on the molecular docking studies, HCA does bind to the active sites of torcetrapib is and based on the MD simulation analysis, the binding of the ligand-protein complexes are stable throughout the simulation. Further verification on in vitro experimentation of HCA does shows the potency against CETP activity and the results are in consonance with the in silico studies. Conclusion: This study prepares a solid foundation on how the binding mode of HCA against CETP and the study of mechanism of actions should be carry forward in order to see the real interaction between CETP protein residue and HCA in increasing the HDL-C level

Preliminary evaluation of CETP inhibition from selected Garcinia species

Two types of Garcinia species which are Garcinia parvifolia and Garcinia atroviridis Griff ex T. Anders were selected and being labelled as UNMC 45L, UNMC 78T and UNMC 78T based on the folklore medicine „myths? that claiming Garcinia species has the ability to be anti-cholesterol. All of these three plant parts were evaluated for therapeutic potential as CETP inhibitors by using CETP Inhibitor drug screening kit. Extraction of crude material from plants was performed via gradient maceration in hexane, ethyl acetate and ethanol. All of the extracts show significant inhibition towards CETP activity. Ethanol extracts of UNMC 45L shows greatest inhibition as the IC50 is 15.43 ± 0.4212 mg/ml followed by Hexane extract and Ethyl Acetate extracts of UNMC 78L which are 28.70 ± 1.320 mg/ml and 28.49 ± 1.126 mg/ml respectively. However, all of the extracts of UNMC 78T shows lowest inhibition towards CETP activity and it is assumed that more bioactive compound could be present in the leaves compare to twigs. The positive findings from this study suggest that Garcinia species was effective natural inhibitors towards CET

Mixed anaerobic fermentation by clostridium and pseudomonas species

The selection of bacterial strains for the anaerobic fermentation if crucial since it determine fermentation performance and influences methods for feedstock pre-treatment, hydrolysis, and solvent recovery. The main bacterial strain used in anaerobic fermentation of ABE production is Clostridium species. However, the performance of wild-type Clostridium strains is limited by solvent tolerance and low cell density during the solventogenic phase of Clostridium growth and causes low production of solvent. So mixed cultures are proposed to overcome the mentioned drawbacks

Lignocellulosic Biomass – A Sustainable Feedstock for Acetone-Butanol-Ethanol Fermentation

Biobutanol has been identified as a promising future biofuel. However, generally the extraction and separation of biobutanol from the fermentation mixture is a costly process. Therefore, the idea of using acetone-butanol-ethanol (ABE) mixture directly as biofuel were proposed to eliminate the recovery process. ABE has been identified as a promising future biofuel. The feedstocks play an important role in the feasibility of ABE as a fuel. Lignocellulosic biomass is seen as a promising feedstock for the production of biofuels. Thus, in this review, ABE biofuel is been summarized from three aspects namely (i) selection of feedstocks, (ii) microbial selection and (iii) hydrolysis, fermentation, and purification techniques. Anaerobic fermentation together with commonly employed recovery processes are discussed in the second part of this review. This review concludes with different challenges and future research in ABE fermentation that can pave the way for future commercialization of this promising biofuel

Preliminary study on bioethanol production from starchy foodwastes by immobilized saccharomyces cerevisiae

Usually, fuels like ethanol and butanol were produced from crude oil and fossil fuels. However, due to increasing fuel demand and limited reserves of fossils, biofuels have caught much attention these days. Biofuels such as bioethanol can be produced from a variety of renewable feedstocks. Food wastes are a promising feedstock since utilization of these can reduce environmental pollution. Thus, in this study starchy food wastes of bread, rice and potatoes were used as a potential feedstock for the production of bioethanol. The Saccharomyces cerevisiae used in this research was at first immobilized in calcium alginate beads through entrapment technique. Later the effect of temperature on bioethanol efficiency was studied using these immobilized yeasts. It was found out that the highest fermentation efficiency of 1.24% was obtained at 30oC. However, more studies are required to optimize the whole process of fermentation of bioethanol from these starchy foodwastes

An investigation of cytokines and chemokines interaction using network analysis in covid-19

The emergence of a pandemic coronavirus disease 2019 (COVID-19) caused by infection with SARS-CoV-2 have become threats to humanity. In terms of their physio pathological pathways, a wide variety of biomolecules have been activated, based on immunological responses. It is therefore relevant to compare the human respiratory cell lines to infections with the SARS-CoV-2 and other respiratory viruses. In this study, we examined gene expression profiles of GSE147507 from the Gene Expression Omnibus (GEO) were used to explore the transcriptional response of SARS-CoV-2 with other respiratory viruses, including human parainfluenza virus 3 (HPIV3), respiratory syncytial virus (RSV), and influenza A virus (IAV), in human respiratory cell lines. Network Analyst 3.0 software was used to perform this gene expression data via intuitive web interface. Through its well-established statistical procedures with state- of-the-art data visualization techniques, it allows us to navigate large complex gene expression data sets to determine important features, patterns, functions and connections that would lead us to a new biological hypothesis. The raw RNA-sequencing data undergoes data processing, including filtering, quality check and normalization before it corresponds to data analysis and interpretation. The edger package was used to identify differentially expressed genes (DEGs) on respiratory viruses infected in human lung epithelium- derived cell lines, such as lung alveolar cells (A549), A549 cells expressing ACE2 (A549-ACE2) and cultured human airway epithelial cells (NHBE). P |2| were set as thresholds for identifying DEGs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were applied for the functional annotation and pathway analysis. Interpretation of gene expression data obtained from the visualization of volcano plot and analysis of protein-protein interaction (PPI) to reveal the functional associations between proteins on a genome-wide scale using STRING interactome

The prevalence of insulin resistance in Malaysia and Indonesia: an updated systematic review and meta-analysis

Noncommunicable diseases such as diabetes are strongly associated with the insulin resistance (IR) status of an individual. However, the prevalence of insulin resistance in Southeast Asia is poorly reported. Hence, this study investigated the prevalence of IR in Southeast Asia from the year 2016 to 2021. This study was carried out according to PRISMA guidelines. The literature search was conducted utilizing the PubMed and SCOPUS databases from the year 2016 to 2021 using the keywords ‘(insulin AND resistance) OR (insulin AND sensitivity) OR (prevalence OR incidence) AND (Malaysia OR Thailand OR Singapore OR Brunei OR Cambodia OR Indonesia OR Laos OR Myanmar OR Philippines OR Timor leste OR Vietnam)’. Funnel plot and publication bias were assessed using Egger’s tests. Data were expressed as the prevalence rate. Results: A total of 12 studies with 2198 subjects were considered in the meta-analysis. Significant heterogeneity (I2 > 94% and p-value < 0.001) was observed in the meta-analysis. The overall prevalence of IR in Southeast Asia was 44.3%, with Malaysia having the highest prevalence rate at 50.4%, followed by Indonesia at 44.2%. Bias was detected in the meta-analysis. It may be that reports published before the year 2016 met the study selection criteria, but were excluded from the meta-analysis. The results from the meta-analysis indicate that the prevalence of IR in Southeast Asia is very high. This provided insights for healthcare policy makers and public health officials in designing IR screening programs

Biofilm formation of serotype 19 Streptococcus pneumoniae clinical isolates in relation to clinical isolate source, pH and Fe(III) supplementation

Streptococcus pneumoniae is one the world’s foremost bacterial pathogens that cause massive global mortality and morbidity in young children and immunocompromised adults especially in developing countries. Biofilms have been increasingly recognized as an important prerequisite to disease. Individual S. pneumoniae strains differ markedly in their virulence phenotypes, but genetic heterogeneity has complicated attempts to identify any association between a given clonal lineage and propensity to cause a particular disease type. This study investigated serotype 19 S. pneumoniae from blood and ear isolates for biofilm formation capacity in relation to isolate source, pH and ferric oxide [Fe(III)] supplementation. Viable count and density biofilm assays, microscopy and multi locus sequence typing (MLST) were applied to investigate biofilm formation capacity and genetic diversity of serotype 19 S. pneumoniae from blood and ear isolates. Generally, blood isolates were observed to produce more biofilms at both pH 7.4 and 6.8 compared to the ear isolates. The supplementation of Fe(III) was also found to support biofilm growth. Upon MLST typing of the isolates, marked differences in biofilm formation within the same sequence types (ST) of ST199 and ST177 was observed. This strongly indicated that strains within the same sequence type show differences in biofilm formation capacity. This study showed that despite belonging to the same serotype, serotype 19, S. pneumoniae blood and ear isolates showed high diversity in biofilm formation ability in relation to pH and Fe(III) supplementation. Additionally, pneumococcal isolates from sequence types ST199 and ST177 also gave rise to differences in biofilm formation ability within the same sequence type (ST). The diversity of biofilm formation within serotype 19 seen in this study is significant to further inform of vaccination strategies against pneumococcal infections, in that due to variations in biofilm formation capacity within the same ST. It is possible that within serotype 19 may show variable vaccination or drug treatment responses. This also indicates that the current treatment strategy which employs specific serotype selection as for PCV14 and PCV7 pneumococcal vaccines may not produce the desired therapeutic results