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A contemporary biological pathway of islet amyloid polypeptide for the management of diabetic dementia

Sah SK, Samuel VP, Dahiya S, Singh Y, Gilhotra RM, Gupta G, et al.

Chem Biol Interact, 2019 Jun 01;306:117-122.
PMID: 31004596 DOI: 10.1016/j.cbi.2019.04.022

Major challenges of dealing elder patients with diabetes mellitus (DM) are the individualization of consideration in persons with various comorbid types of conditions. In spite of the fact that microvascular and macrovascular problems associated with DM are well documented, there is only a few numbers of reports viewing different conditions, for example, cognitive dysfunction. Cognitive dysfunction is of specific significance due to its effect on self-care and quality of life. All in all, the etiology of cognitive dysfunction in the maturing populace is probably going to be the grouping of ischemic and degenerative pathology. It is likewise trusted that Hyperglycemia is engaged with the system of DM-related cognitive dysfunction. At present, it isn't certain in the case of enhancing glycemic control or utilizing therapeutic agents can enhance the risk of cognitive decay. Amylin was later characterized as an amyloidogenic peptide, confined from a beta cell tumor and called islet amyloid polypeptide (IAPP), and after that, amylin. Conversely, we investigate the beneficial role and hypothesizing the mechanism of amylin related expanding the level and activation of CGRP receptor to enhance the cognition declination amid diabetic dementia.

Matched MeSH terms: Diabetes Mellitus/metabolism*; Islets of Langerhans/metabolism; Islet Amyloid Polypeptide/metabolism*
Telomerase reverse transcriptase downregulation by RNA interference modulates endoplasmic reticulum stress and mitochondrial energy production

Mohd Zain MZ, Ismail NH, Ahmad N, Sulong S, Karsani SA, Abdul Majid N

Mol Biol Rep, 2020 Oct;47(10):7735-7743.
PMID: 32959195 DOI: 10.1007/s11033-020-05848-y

Telomerase is a cancer promoting ribonucleoprotein complex and is a potential therapeutic target for cancer. In this study, the effects of telomerase downregulation on the whole cell proteome were investigated. Understanding how the effect of downregulation on the whole proteome profile will generate a greater understanding of the possible roles played by telomerase in cancer. Downregulation was achieved by RNA interference (RNAi), targeting the telomerase reverse transcriptase (TERT) subunits of telomerase. Transfection of TERT siRNA downregulates TERT gene expression and induced downregulation of telomerase activity. Investigation of the effect of silencing TERT in telomerase was further validated through proteomic analysis by performing 2-dimension electrophoresis (2DE) coupled with MALDI-TOF/TOF. 12 protein spots in HeLa cells were reported to be significantly differentially expressed with 11 of them were upregulated and 1 downregulated. Through STRING analysis, differentially expressed proteins demonstrated strong associations with endoplasmic reticulum stress marker and mitochondrial energy production marker. In conclusions, the result exhibited novel integrated proteomic response involving endoplasmic reticulum stress and mitochondrial energy production in response to the TERT downregulation in cervical cancer cells.

Matched MeSH terms: Energy Metabolism*; Mitochondria/metabolism*
Protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 for catalytic efficiency improvement on kenaf biomass hydrolysis

Damis SIR, Murad AMA, Diba Abu Bakar F, Rashid SA, Jaafar NR, Illias RM

Enzyme Microb Technol, 2019 Dec;131:109383.
PMID: 31615675 DOI: 10.1016/j.enzmictec.2019.109383

Enzyme hydrolysis faces a bottleneck due to the recalcitrance of the lignocellulose biomass. The protein engineering of GH11 xylanase from Aspergillus fumigatus RT-1 was performed near the active site and at the N-terminal region to improve its catalytic efficiency towards pretreated kenaf (Hibiscus cannabinus) hydrolysis. Five mutants were constructed by combined approaches of error-prone PCR, site-saturation and site-directed mutagenesis. The double mutant c168 t/Q192H showed the most effective hydrolysis reaction with a 13.9-fold increase in catalytic efficiency, followed by mutants Y7L and c168 t/Q192 H/Y7L with a 1.6-fold increase, respectively. The enhanced catalytic efficiency evoked an increase in sugar yield of up to 28% from pretreated kenaf. In addition, mutant c168 t/Q192 H/Y7L improved the thermostability at higher temperature and acid stability. This finding shows that mutations at distances less than 15 Å from the active site and at putative secondary binding sites affect xylanase catalytic efficiency towards insoluble substrates hydrolysis.

Matched MeSH terms: Xylosidases/metabolism*; Hibiscus/metabolism*; Mutant Proteins/metabolism
Crystal structure of a compact α-amylase from Geobacillus thermoleovorans

Mok SC, Teh AH, Saito JA, Najimudin N, Alam M

Enzyme Microb Technol, 2013 Jun 10;53(1):46-54.
PMID: 23683704 DOI: 10.1016/j.enzmictec.2013.03.009

A truncated form of an α-amylase, GTA, from thermophilic Geobacillus thermoleovorans CCB_US3_UF5 was biochemically and structurally characterized. The recombinant GTA, which lacked both the N- and C-terminal transmembrane regions, functioned optimally at 70°C and pH 6.0. While enzyme activity was not enhanced by the addition of CaCl2, GTA's thermostability was significantly improved in the presence of CaCl2. The structure, in complex with an acarbose-derived pseudo-hexasaccharide, consists of the typical three domains and binds one Ca(2+) ion. This Ca(2+) ion was strongly bound and not chelated by EDTA. A predicted second Ca(2+)-binding site, however, was disordered. With limited subsites, two novel substrate-binding residues, Y147 and Y182, may help increase substrate affinity. No distinct starch-binding domain is present, although two regions rich in aromatic residues have been observed. GTA, with a smaller domain B and several shorter loops compared to other α-amylases, has one of the most compact α-amylase folds that may contribute greatly to its tight Ca(2+) binding and thermostability.

Matched MeSH terms: alpha-Amylases/metabolism*; Calcium/metabolism; Acarbose/metabolism
Dendrimer-like nanoparticles based β-galactosidase assembly for enhancing its selectivity toward transgalactosylation

Misson M, Du X, Jin B, Zhang H

Enzyme Microb Technol, 2016 Mar;84:68-77.
PMID: 26827776 DOI: 10.1016/j.enzmictec.2015.12.008

Functional nanomaterials have been pursued to assemble nanobiocatalysts since they can provide unique hierarchical nanostructures and localized nanoenvironments for enhancing enzyme specificity, stability and selectivity. Functionalized dendrimer-like hierarchically porous silica nanoparticles (HPSNs) was fabricated for assembling β-galactosidase nanobiocatalysts for bioconversion of lactose to galacto-oligosaccharides (GOS). The nanocarrier was functionalized with amino (NH2) and carboxyl (COOH) groups to facilitate enzyme binding, benchmarking with non-functionalized HPSNs. Successful conjugation of the functional groups was confirmed by FTIR, TGA and zeta potential analysis. HPSNs-NH2 showed 1.8-fold and 1.1-fold higher β-galactosidase adsorption than HPSNs-COOH and HPSNs carriers, respectively, with the highest enzyme adsorption capacity of 328mg/g nanocarrier at an initial enzyme concentration of 8mg/ml. The HPSNs-NH2 and β-galactosidase assembly (HPSNs-NH2-Gal) demonstrated to maintain the highest activity at all tested enzyme concentrations and exhibited activity up to 10 continuous cycles. Importantly, HPSNs-NH2-Gal was simply recycled through centrifugation, overcoming the challenging problems of separating the nanocarrier from the reaction medium. HPSNs-NH2-Gal had distinguished catalytic reaction profiles by favoring transgalactosylation, enhancing GOS production of up to 122g/l in comparison with 56g/l by free β-galactosidase. Furthermore, it generated up to 46g/l GOS at a lower initial lactose concentration while the free counterpart had negligible GOS production as hydrolysis was overwhelmingly dominant in the reaction system. Our research findings show the amino-functionalized HPSNs can selectively promote the enzyme activity of β-galactosidase for transgalactosylation, which is beneficial for GOS production.

Matched MeSH terms: beta-Galactosidase/metabolism*; Enzymes, Immobilized/metabolism; Lactose/metabolism
Comparative Transcriptome Analysis Provides Insights into Differentially Expressed Genes and Long Non-Coding RNAs between Ovary and Testis of the Mud Crab (Scylla paramamosain)

Yang X, Ikhwanuddin M, Li X, Lin F, Wu Q, Zhang Y, et al.

Mar Biotechnol (NY), 2018 Feb;20(1):20-34.
PMID: 29152671 DOI: 10.1007/s10126-017-9784-2

The molecular mechanism underlying sex determination and gonadal differentiation of the mud crab (Scylla paramamosain) has received considerable attention, due to the remarkably biological and economic differences between sexes. However, sex-biased genes, especially non-coding genes, which account for these differences, remain elusive in this crustacean species. In this study, the first de novo gonad transcriptome sequencing was performed to identify both differentially expressed genes and long non-coding RNAs (lncRNAs) between male and female S. paramamosain by using Illumina Hiseq2500. A total of 79,282,758 and 79,854,234 reads were generated from ovarian and testicular cDNA libraries, respectively. After filtrating and de novo assembly, 262,688 unigenes were produced from both libraries. Of these unigenes, 41,125 were annotated with known protein sequences in public databases. Homologous genes involved in sex determination and gonadal development pathways (Sxl-Tra/Tra-2-Dsx/Fru, Wnt4, thyroid hormone synthesis pathway, etc.) were identified. Three hundred and sixteen differentially expressed unigenes were further identified between both transcriptomes. Meanwhile, a total of 233,078 putative lncRNAs were predicted. Of these lncRNAs, 147 were differentially expressed between sexes. qRT-PCR results showed that nine lncRNAs negatively regulated the expression of eight genes, suggesting a potential role in sex differentiation. These findings will provide fundamental resources for further investigation on sex differentiation and regulatory mechanism in crustaceans.

Matched MeSH terms: Brachyura/metabolism*; Ovary/metabolism*; Testis/metabolism*
Discovery of new inhibitor for the protein arginine deiminase type 4 (PAD4) by rational design of α-enolase-derived peptides

Ahmad Nadzirin I, Chor ALT, Salleh AB, Rahman MBA, Tejo BA

Comput Biol Chem, 2021 Jun;92:107487.
PMID: 33957477 DOI: 10.1016/j.compbiolchem.2021.107487

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease affecting about 0.24 % of the world population. Protein arginine deiminase type 4 (PAD4) is believed to be responsible for the occurrence of RA by catalyzing citrullination of proteins. The citrullinated proteins act as autoantigens by stimulating an immune response. Citrullinated α-enolase has been identified as one of the autoantigens for RA. Hence, α-enolase serves as a suitable template for design of potential peptide inhibitors against PAD4. The binding affinity of α-enolase-derived peptides and PAD4 was virtually determined using PatchDock and HADDOCK docking programs. Synthesis of the designed peptides was performed using a solid phase peptide synthesis method. The inhibitory potential of each peptide was determined experimentally by PAD4 inhibition assay and IC50 measurement. PAD4 assay data show that the N-P2 peptide is the most favourable substrate among all peptides. Further modification of N-P2 by changing the Arg residue to canavanine [P2 (Cav)] rendered it an inhibitor against PAD4 by reducing the PAD4 activity to 35 % with IC50 1.39 mM. We conclude that P2 (Cav) is a potential inhibitor against PAD4 and can serve as a starting point for the development of even more potent inhibitors.

Matched MeSH terms: Enzyme Inhibitors/metabolism; Peptides/metabolism; Phosphopyruvate Hydratase/metabolism
4-chloro-1,2-phenylenediamine induces apoptosis in Mardin-Darby canine kidney cells via activation of caspases

Onn LC, Ching CS, Lian TY, Foon LV, Chew Hee N, Moi CS

Environ Toxicol, 2014 Jun;29(6):655-64.
PMID: 22778066 DOI: 10.1002/tox.21792

4-Chloro-1,2-phenylenediamine (4-Cl-o-PD) is a halogenated aromatic diamine that was used as a precursor for manufacturing permanent hair dyes. Despite its well-documented mutagenic and carcinogenic effects in a number of in vitro and in vivo models, its cytotoxicity and mode of action have not received similar attention. Here, we investigated the effect of 4-Cl-o-PD on Mardin-Darby canine kidney cells. It induced apoptosis and the evidence suggests its initiation by reactive oxygen species (ROS). The results of various assays used show a dose-dependent (i) decrease in cell viability, (ii) increase in cells at sub-G1 phase and the G0/G1 phase arrested in cell cycle, (iii) increase in intracellular ROS accompanied by depletion of glutathione, and (iv) that apoptotic cell death probably involves activation of both intrinsic and extrinsic pathways.

Matched MeSH terms: Glutathione/metabolism; Reactive Oxygen Species/metabolism; Caspases/metabolism*
Establishment of epithelial and fibroblast cell cultures and cell lines from primary renal cancer nephrectomies

Yap NY, Ong TA, Morais C, Pailoor J, Gobe GC, Rajandram R

Cell Biol Int, 2019 Jun;43(6):715-725.
PMID: 31062478 DOI: 10.1002/cbin.11150

Renal cell carcinoma (RCC) is one of the most lethal urogenital cancers and effective treatment of metastatic RCC remains an elusive target. Cell lines enable the in vitro investigation of molecular and genetic changes leading to renal carcinogenesis and are important for evaluating cellular drug response or toxicity. This study details a fast and easy protocol of establishing epithelial and fibroblast cell cultures or cell lines concurrently from renal cancer nephrectomy tissue. The protocol involves mechanical disaggregation, collagenase digestion and cell sieving for establishing epithelial cells while fibroblast cells were grown from explants. This protocol has been modified from previous published reports with additional antibiotics and washing steps added to eliminate microbial contamination from the surgical source. Cell characterisation was carried out using immunofluorescence and quantitative polymerase chain reaction. Eleven stable epithelial renal tumour cell lines of various subtypes, including rare subtypes, were established with a spontaneous immortalisation rate of 21.6% using this protocol. Eight fibroblast cell cultures grew successfully but did not achieve spontaneous immortalisation. Cells of epithelial origin expressed higher expressions of epithelial markers such as pan-cytokeratin, cytokeratin 8 and E-cadherin whereas fibroblast cells expressed high α-smooth muscle actin. Further mutational analysis is needed to evaluate the genetic or molecular characteristics of the cell lines.

Matched MeSH terms: Carcinoma, Renal Cell/metabolism; Epithelial Cells/metabolism; Fibroblasts/metabolism
Fulltext Modulation of Crustacean Innate Immune Response by Amino Acids and Their Metabolites: Inferences From Other Species

Huang Z, Aweya JJ, Zhu C, Tran NT, Hong Y, Li S, et al.

Front Immunol, 2020;11:574721.
PMID: 33224140 DOI: 10.3389/fimmu.2020.574721

Aquaculture production of crustaceans (mainly shrimp and crabs) has expanded globally, but disease outbreaks and pathogenic infections have hampered production in the last two decades. As invertebrates, crustaceans lack an adaptive immune system and mainly defend and protect themselves using their innate immune system. The immune system derives energy and metabolites from nutrients, with amino acids constituting one such source. A growing number of studies have shown that amino acids and their metabolites are involved in the activation, synthesis, proliferation, and differentiation of immune cells, as well as in the activation of immune related signaling pathways, reduction of inflammatory response and regulation of oxidative stress. Key enzymes in amino acid metabolism have also been implicated in the regulation of the immune system. Here, we reviewed the role played by amino acids and their metabolites in immune-modulation in crustaceans. Information is inferred from mammals and fish where none exists for crustaceans. Research themes are identified and the relevant research gaps highlighted for further studies.

Matched MeSH terms: Amino Acids/metabolism; Crustacea/metabolism; Immune System/metabolism
Preliminary examination of microRNA expression profiling in bipolar disorder I patients during antipsychotic treatment

Lim CH, Zainal NZ, Kanagasundram S, Zain SM, Mohamed Z

Am J Med Genet B Neuropsychiatr Genet, 2016 09;171(6):867-74.
PMID: 27177356 DOI: 10.1002/ajmg.b.32457

Although major progress has been achieved in research and development of antipsychotic medications for bipolar disorder (BPD), knowledge of the molecular mechanisms underlying this disorder and the action of atypical antipsychotics remains incomplete. The levels of microRNAs (miRNAs)-small non-coding RNA molecules that regulate gene expression, including genes involved in neuronal function and plasticity-are frequently altered in psychiatric disorders. This study aimed to examine changes in miRNA expression in bipolar mania patients after treatment with asenapine and risperidone. Using a miRNA microarray, we analyzed miRNA expression in the blood of 10 bipolar mania patients following 12 weeks of treatment with asenapine or risperidone. Selected miRNAs were validated by using real-time PCR. A total of 16 miRNAs were differentially expressed after treatment in the asenapine group, 14 of which were significantly upregulated and the other two significantly downregulated. However, all three differentially expressed miRNAs in the risperidone group were downregulated. MiRNA target gene prediction and gene ontology analysis revealed significant enrichment for pathways associated with immune system response and regulation of programmed cell death and transcription. Our results suggest that candidate miRNAs may be involved in the mechanism of action of both antipsychotics in bipolar mania. © 2016 Wiley Periodicals, Inc.

Matched MeSH terms: Bipolar Disorder/metabolism; Antipsychotic Agents/metabolism; MicroRNAs/metabolism*
Fulltext Surface display of glycosylated Tyrosinase related protein-2 (TRP-2) tumour antigen on Lactococcus lactis

Kalyanasundram J, Chia SL, Song AA, Raha AR, Young HA, Yusoff K

BMC Biotechnol, 2015;15:113.
PMID: 26715153 DOI: 10.1186/s12896-015-0231-z

The exploitation of the surface display system of food and commensal lactic acid bacteria (LAB) for bacterial, viral, or protozoan antigen delivery has received strong interest recently. The Generally Regarded as Safe (GRAS) status of the Lactococcus lactis coupled with a non-recombinant strategy of in-trans surface display, provide a safe platform for therapeutic drug and vaccine development. However, production of therapeutic proteins fused with cell-wall anchoring motifs is predominantly limited to prokaryotic expression systems. This presents a major disadvantage in the surface display system particularly when glycosylation has been recently identified to significantly enhance epitope presentation. In this study, the glycosylated murine Tyrosinase related protein-2 (TRP-2) with the ability to anchor onto the L. lactis cell wall was produced in suspension adapted Chinese Hamster Ovary (CHO-S) cells by expressing TRP-2 fused with cell wall anchoring LysM motif (cA) at the C-terminus.

Matched MeSH terms: Cell Wall/metabolism*; Lactococcus lactis/metabolism*; Intramolecular Oxidoreductases/metabolism*
Fulltext Evaluation of insulin expression and secretion in genetically engineered gut K and L-cells

Ahmad Z, Rasouli M, Azman AZ, Omar AR

BMC Biotechnol, 2012 Sep 19;12:64.
PMID: 22989329 DOI: 10.1186/1472-6750-12-64

BACKGROUND: Gene therapy could provide an effective treatment of diabetes. Previous studies have investigated the potential for several cell and tissue types to produce mature and active insulin. Gut K and L-cells could be potential candidate hosts for gene therapy because of their special features.
RESULTS: In this study, we isolated gut K and L-cells to compare the potential of both cell types to produce insulin when exposed to similar conditions. The isolated pure K and L-cells were transfected with recombinant plasmids encoding insulin and with specific promoters for K or L-cells. Insulin expression was studied in response to glucose or meat hydrolysate. We found that glucose and meat hydrolysate efficiently induced insulin secretion from K and L-cells. However, the effects of meat hydrolysate on insulin secretion were more potent in both cells compared with glucose. Results of enzyme-linked immunosorbent assays showed that L-cells secreted more insulin compared with K-cells regardless of the stimulator, although this difference was not statistically significant.
CONCLUSION: The responses of K and L-cells to stimulation with glucose or meat hydrolysate were generally comparable. Therefore, both K and L-cells show similar potential to be used as surrogate cells for insulin gene expression in vitro. The potential use of these cells for diabetic gene therapy warrants further investigation.

Matched MeSH terms: Glucose/metabolism; Insulin/metabolism*; Intestinal Mucosa/metabolism
Biosolids accumulation and biodegradation of domestic wastewater treatment plant sludge by developed liquid state bioconversion process using a batch fermenter

Alam MZ, Fakhru'l-Razi A, Molla AH

Water Res, 2003 Sep;37(15):3569-78.
PMID: 12867323

The biosolids accumulation and biodegradation of domestic wastewater treatment plant (DWTP) sludge by filamentous fungi have been investigated in a batch fermenter. The filamentous fungi Aspergillus niger and Penicillium corylophilum isolated from wastewater and DWTP sludge was used to evaluate the treatment performance. The optimized mixed inoculum (A. niger and P. corylophilum) and developed process conditions (co-substrate and its concentration, temperature, initial pH, inoculum size, and aeration and agitation rate) were incorporated to accelerate the DWTP sludge treatment process. The results showed that microbial treatment of higher strength of DWTP sludge (4% w/w of TSS) was highly influenced by the liquid state bioconversion (LSB) process. In developed bioconversion processes, 93.8 g/kg of biosolids was enriched with fungal biomass protein of 30 g/kg. Enrichment of nutrients such as nitrogen (N), phosphorous (P), potassium (K) in biosolids was recorded in 6.2% (w/w), 3.1% (w/w) and 0.15% (w/w) from its initial values of 4.8% (w/w), 2.0% (w/w) and 0.08% (w/w) respectively after 10 days of fungal treatment. The biodegradation results revealed that 98.8% of TSS, 98.2% of TDS, 97.3% of turbidity, 80.2% of soluble protein, 98.8% of reducing sugar and 92.7% of COD in treated DWTP sludge supernatant were removed after 8 days of microbial treatment. The specific resistance to filtration (SRF) in treated sludge (1.4x10(12) m/kg) was decreased tremendously by the microbial treatment of DWTP sludge after 6 days of fermentation compared to untreated sample (85x10(12) m/kg).

Matched MeSH terms: Nitrogen/metabolism; Phosphorus/metabolism; Proteins/metabolism
The use of response surface methodology for enhanced production of a thermostable bacterial lipase in a novel yeast system

Abu ML, Mohammad R, Oslan SN, Salleh AB

Prep Biochem Biotechnol, 2021;51(4):350-360.
PMID: 32940138 DOI: 10.1080/10826068.2020.1818256

A thermostable bacterial lipase from Geobacillus zalihae was expressed in a novel yeast Pichia sp. strain SO. The preliminary expression was too low and discourages industrial production. This study sought to investigate the optimum conditions for T1 lipase production in Pichia sp. strain SO. Seven medium conditions were investigated and optimized using Response Surface Methodology (RSM). Five responding conditions namely; temperature, inoculum size, incubation time, culture volume and agitation speed observed through Plackett-Burman Design (PBD) method had a significant effect on T1 lipase production. The medium conditions were optimized using Box-Behnken Design (BBD). Investigations reveal that the optimum conditions for T1 lipase production and Biomass concentration (OD600) were; Temperature 31.76 °C, incubation time 39.33 h, culture volume 132.19 mL, inoculum size 3.64%, and agitation speed of 288.2 rpm with a 95% PI low as; 12.41 U/mL and 95% PI high of 13.65 U/mL with an OD600 of; 95% PI low as; 19.62 and 95% PI high as; 22.62 as generated by the software was also validated. These predicted parameters were investigated experimentally and the experimental result for lipase activity observed was 13.72 U/mL with an OD600 of 24.5. At these optimum conditions, there was a 3-fold increase on T1 lipase activity. This study is the first to develop a statistical model for T1 lipase production and biomass concentration in Pichia sp. Strain SO. The optimized production of T1 lipase presents a choice for its industrial application.

Matched MeSH terms: Methanol/metabolism; Culture Media/metabolism; Pichia/metabolism*
Effects of feeding fermented fish on egg cholesterol content in hens

Loh TC, Law FL, Goh YM, Foo HL, Zulkifli I

Anim Sci J, 2009 Feb;80(1):27-33.
PMID: 20163464 DOI: 10.1111/j.1740-0929.2008.00591.x

This study was conducted to investigate the effects of feeding fermented fish (FF) to layers on laying performance, and polyunsaturated fatty acid and cholesterol levels in eggs and plasma. A total of 96, 13-week-old Babcock B380 pullets were used in this study. They were randomly assigned to four numerically equal groups with eight replicates per treatment, three birds per replicate. All the birds were housed in individual cages. The dietary treatments were: Control diet, without FF; FF3 diet containing 3% (w/w) FF, FF6 diet containing 6% (w/w) FF and FF9 diet containing 9% (w/w) FF. The study was carried out for 16 weeks inclusive of two weeks of adjustment. Weekly feed intake and egg production were recorded. Blood plasma cholesterol and fatty acid profiles were assayed at the end of the experiment. FF did not enhance (P > 0.05) egg mass but (P < 0.05) decreased egg weight slightly. However, egg yolk cholesterol and plasma cholesterol concentrations were reduced (P < 0.05) by FF. The n-6:n-3 fatty acids ratio in the egg yolk (Control = 7.9, FF9 = 6.2) and plasma (Control = 10.6, FF9 = 6.2) were decreased by feeding FF. Moreover, FF was able to increase (P < 0.05) the docosahexaenoic acid (DHA) concentrations in egg yolk and plasma. In conclusion, this study demonstrated that FF increased DHA and reduced egg yolk cholesterol in poultry eggs.

Matched MeSH terms: Cholesterol/metabolism; Docosahexaenoic Acids/metabolism; Egg Yolk/metabolism
Cloning and functional analysis of the genes coding for 4-aminobenzenesulfonate 3,4-dioxygenase from Hydrogenophaga sp. PBC

Gan HM, Shahir S, Yahya A

Microbiology (Reading), 2012 Aug;158(Pt 8):1933-1941.
PMID: 22609751 DOI: 10.1099/mic.0.059550-0

The gene coding for the oxygenase component, sadA, of 4-aminobenzenesulfonate (4-ABS) 3,4-dioxygenase in Hydrogenophaga sp. PBC was previously identified via transposon mutagenesis. Expression of wild-type sadA in trans restored the ability of the sadA mutant to grow on 4-ABS. The inclusion of sadB and sadD, coding for a putative glutamine-synthetase-like protein and a plant-type ferredoxin, respectively, further improved the efficiency of 4-ABS degradation. Transcription analysis using the gfp promoter probe plasmid showed that sadABD was expressed during growth on 4-ABS and 4-sulfocatechol. Heterologous expression of sadABD in Escherichia coli led to the biotransformation of 4-ABS to a metabolite which shared a similar retention time and UV/vis profile with 4-sulfocatechol. The putative reductase gene sadC was isolated via degenerate PCR and expression of sadC and sadABD in E. coli led to maximal 4-ABS biotransformation. In E. coli, the deletion of sadB completely eliminated dioxygenase activity while the deletion of sadC or sadD led to a decrease in dioxygenase activity. Phylogenetic analysis of SadB showed that it is closely related to the glutamine-synthetase-like proteins involved in the aniline degradation pathway. This is the first discovery, to our knowledge, of the functional genetic components for 4-ABS aromatic ring hydroxylation in the bacterial domain.

Matched MeSH terms: Bacterial Proteins/metabolism; Sulfanilic Acids/metabolism*; Dioxygenases/metabolism
Fulltext The PCA and LDA analysis on the differential expression of proteins in breast cancer

Liang S, Singh M, Dharmaraj S, Gam LH

Dis Markers, 2010;29(5):231-42.
PMID: 21206008 DOI: 10.3233/DMA-2010-0753

Breast cancer is a leading cause of mortality in women. In Malaysia, it is the most common cancer to affect women. The most common form of breast cancer is infiltrating ductal carcinoma (IDC). A proteomic approach was undertaken to identify protein profile changes between cancerous and normal breast tissues from 18 patients. Two protein extracts; aqueous soluble and membrane associated protein extracts were studied. Thirty four differentially expressed proteins were identified. The intensities of the proteins were used as variables in PCA and reduced data of six principal components (PC) were subjected to LDA in order to evaluate the potential of these proteins as collective biomarkers for breast cancer. The protein intensities of SEC13-like 1 (isoform b) and calreticulin contributed the most to the first PC while the protein intensities of fibrinogen beta chain precursor and ATP synthase D chain contributed the most to the second PC. Transthyretin precursor and apolipoprotein A-1 precursor contributed the most to the third PC. The results of LDA indicated good classification of samples into normal and cancerous types when the first 6 PCs were used as the variables. The percentage of correct classification was 91.7% for the originally grouped tissue samples and 88.9% for cross-validated samples.

Matched MeSH terms: Breast Neoplasms/metabolism*; Carcinoma, Ductal, Breast/metabolism*; Proteome/metabolism
Fulltext The differential expression of aqueous soluble proteins in breast normal and cancerous tissues in relation to ethnicity of the patients; Chinese, Malay and Indian

Liang S, Singh M, Gam LH

Dis Markers, 2010;28(3):149-65.
PMID: 20534901 DOI: 10.3233/DMA-2010-0694

Female breast cancer is one of the leading causes of female mortality worldwide. In Malaysia, breast cancer is the most commonly diagnosed cancer in women. Of the women in Malaysia, the Chinese have the highest number of breast cancer cases, followed by the Indian and the Malay. The most common type of breast cancer is infiltrating ductal carcinoma (IDC). A proteomic approach was applied in this study to identify changes in the protein profile of cancerous tissues compared with normal tissues from 18 patients; 8 Chinese, 6 Malay and 4 Indian were analysed. Twenty-four differentially expressed hydrophilic proteins were identified. We evaluated the potential of these proteins as biomarkers for infiltrating ductal carcinoma based on their ethnic-specific expressions. Three of the upregulated proteins, calreticulin, 14-3-3 protein zeta and 14-3-3 protein eta, were found to be expressed at a significantly higher level in the cancerous breast tissues when compared with the normal tissues in cases of infiltrating ductal carcinoma. The upregulation in expression was particularly dominant in the Malay cohort.

Matched MeSH terms: Breast/metabolism*; Neoplasm Proteins/metabolism*; Proteins/metabolism*
Genetics and evidence for an esterase-associated mechanism of resistance to indoxacarb in a field population of diamondback moth (Lepidoptera: Plutellidae)

Sayyed AH, Wright DJ

Pest Manag Sci, 2006 Nov;62(11):1045-51.
PMID: 16886171

Bioassays (at generation G2) with a newly collected field population (designated CH3) of Plutella xylostella L. from farmers' fields in the Cameron Highlands, Malaysia, indicated resistance ratios of 813-, 79-, 171-, 498- and 1285-fold for indoxacarb, fipronil, spinosad, deltamethrin and Bacillus thuringiensis toxin Cry1Ac respectively compared with a laboratory susceptible population (Lab-UK). At G2 the field-derived population was divided into two subpopulations: one was selected (G2 to G7) with indoxacarb (indoxa-SEL), while the second was left unselected (UNSEL). A significant reduction in the resistance ratio for each compound was observed in UNSEL at G8. For indoxa-SEL, bioassays at G8 found that selection with indoxacarb gave a resistance ratio of 2594 compared with Lab-UK and of 90 compared with UNSEL. The toxicity of fipronil, spinosad and deltamethrin was not significantly different in indoxa-SEL at G8 compared with G2 but was significantly greater than UNSEL at G8. The toxicity of Cry1Ac was significantly reduced in indoxa-SEL at G8 compared with G2 but was also significantly greater than UNSEL at G8. This suggests that indoxacarb selection maintained resistance to these compounds in the indoxa-SEL population. Synergist studies indicated that resistance to indoxacarb in indoxa-SEL was esterase associated. Logit regression analysis of F1 reciprocal crosses between indoxa-SEL and Lab-UK indicated that resistance to indoxacarb was inherited as an autosomal, incompletely recessive (D(LC) = 0.35) trait. Tests of monogenic inheritance suggested that resistance to indoxacarb was controlled by a single locus.

Matched MeSH terms: Esterases/metabolism*; Insecticides/metabolism*; Oxazines/metabolism*

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