Chemometric approach to validating faecal sterols as source tracer for faecal contamination in water

Faecal sterols detection is a promising method for identifying sources of faecal pollution. In this study, faecal contamination in water samples from point source (sewage treatment plants, chicken farms, quail farms and horse stables) was extracted using the solid phase extraction (SPE) technique. Faecal sterols (coprostanol, cholesterol, stigmasterol, β-sitosterol and stigmastanol) were selected as parameters to differentiate the source of faecal pollution. The results indicated that coprostanol, cholesterol and β-sitosterol were the most significant parameters that can be used as source tracers for faecal contamination. Chemometric techniques, such as cluster analysis, principal component analysis and discriminant analysis were applied to the data set on faecal contamination in water from various pollution sources in order to validate the faecal sterols’ profiles. Cluster analysis generated three clusters: coprostanol was in cluster 1, cholesterol and β-sitosterol formed cluster 2, while cluster 3 contained stigmasterol and stigmastanol. Discriminant analysis suggested that coprostanol, cholesterol and β-sitosterol were the most significant parameters to discriminate between the faecal pollution source. The use of chemometric techniques provides useful and promising indicators in tracing the source of faecal contamination.     

Electrical and mechanical properties of carbon pellets from acid (HNO3)treated self-adhesive carbon grain from oil palm empty fruit bunch

Factors such as its natural properties, low cost and availability in large quantities as a by-product may give oil palm empty fruit bunches (EFB) a great potential as a candidate precursor for solid carbon products. Self-adhesive carbon grains (SACG) were prepared from EFB by a low temperature pre-carbonization process. Green pellets were prepared from SACG and SACG treated with nitric acid having concentration of 1, 3 and 5 Molar (M). Carbon pellets were produced by carbonization of green pellets up to 1000°C in a nitrogen environment using multi-step heating profile. Measurements on carbon pellets show that the electrical conductivity (), hardness (H) and Young's modulus (Y) follow the linear equations /[10^–3 × (m )^–1] = 0.57M + 4.74, H/[Vickers hardness] = 27M + 148 and Y/[GPa] = 2.6M + 5.0, respectively. This indicates that nitric acid systematically affected the properties of the product. The behavior that , H and Y increase linearly with M seems to be associated with the effect of acid treatment on the weight loss, lignocellulosic structure and particle size of the SACG.     

Isolation of the MNN9 gene of Yarrowia lipolytica (YlMNN9) and phenotype analysis of a mutant ylmnn9 Delta strain

In this work we describe the isolation of the Yarrowia lipolytica homologue of Saccharomyces cerevisiae MNN9 gene, which we have named YlMNN9, and the phenotype analysis of a Y. lipolytica strain containing the disrupted YlMNN9 allele. YlMNN9 was cloned using degenerate consensus oligonucleotides to generate specific probes that were in turn used to screen mini-gene libraries. The gene is defined by a 1014 bp ORF predicted to encode a protein 337 amino acids long that shares significant homology with the Mnn9ps of S. cerevisiae, Candida albicans and Hansenula polymorpha, including a putative N-terminal transmembrane domain. Disruption of YlMNN9 leads to phenotypes such as resistance to sodium orthovanadate and sensitivity to hygromycin B, compatible with a glycosylation defect, and hypersensitivity to Calcofluor white, Congo red or zymolyase, characteristic of cell wall defects. Analysis of cell wall proteins present in beta-mercaptoethanol and zymolyase extracts showed significant differences between the parental and the ylmnn9 Delta strain. These results suggest that, as has been the case with the mnn9 strain of S. cerevisiae, the ylmnn9 Delta strain we present in this work, could be used to study the cell wall proteins of Y. lipolytica and how they are organized into the cell wall.     

Cauliflower-like poly(3,4-ethylenedioxythipohene)/nanocrystalline cellulose/manganese oxide ternary nanocomposite for supercapacitor

A cauliflower-like ternary nanocomposite of poly(3,4-ethylenedioxythipohene)/nanocrystalline cellulose/manganese oxide (PEDOT/NCC/MnO₂) was synthesized using one-step electropolymerization technique. The effect of manganese (Mn) concentration on the supercapacitive performance was investigated. The structural and morphology studies were conducted using field emission scanning electron microscope, Fourier transform infrared spectroscopy, Raman spectroscopy, and X-ray diffraction. The morphology of ternary nanocomposite at an optimized concentration of Mn resembles the cauliflower-like structure. The two-electrode electrochemical analysis of a ternary nanocomposite PEDOT/NCC/MnO₂ exhibited a higher specific capacitance of 144.69 F/g at 25 mV/s in 1.0 M potassium chloride compared to PEDOT/NCC(63.57 F/g). PEDOT/NCC/MnO₂ ternary nanocomposite was able to deliver a specific power of 494.9 W/kg and 10.3 Wh/kg of specific energy at 1 A g⁻¹ and retained 83% of initial capacitance after 2,000 cycles. These promising results from the incorporation of Mn displayed great prospective in developing PEDOT/NCC/MnO₂ as an electrode material for supercapacitor.     

A Systematic Approach to Assess the Activity and Classification of PCSK9 Variants

Background: Gain of function (GOF) mutations of PCSK9 cause autosomal dominant familial hypercholesterolemia as they reduce the abundance of LDL receptor (LDLR) more efficiently than wild-type PCSK9. In contrast, PCSK9 loss of function (LOF) variants are associated with a hypocholesterolemic phenotype. Dozens of PCSK9 variants have been reported, but most remain of unknown significance since their characterization has not been conducted. Objective: Our aim was to make the most comprehensive assessment of PCSK9 variants and to determine the simplest approach for the classification of these variants. Methods: The expression, maturation, secretion, and activity of nine well-established PCSK9 variants were assessed in transiently transfected HEK293 cells by Western blot and flow cytometry. Their extracellular activities were determined in HepG2 cells incubated with the purified recombinant PCSK9 variants. Their binding affinities toward the LDLR were determined by solid-phase immunoassay. Results: LDLR expression increased when cells were transfected with LOF variants and reduced when cells were transfected with GOF variants compared with wild-type PCSK9. Extracellular activities measurements yielded exactly similar results. GOF and LOF variants had increased, respectively reduced, affinities for the LDLR compared with wild-type PCSK9 with the exception of one GOF variant (R218S) that showed complete resistance to inactivation by furin. All variants were expressed at similar levels and underwent normal maturation and secretion patterns except for two LOF and two GOF mutants. Conclusions: We propose that transient transfections of HEK293 cells with a plasmid encoding a PCSK9 variant followed by LDLR expression assessment by flow cytometry is sufficient to reliably determine its GOF or LOF status. More refined experiments should only be used to determine the underlying mechanism(s) at hand.     

Polyethyleneimine-mediated flocculation of Shewanella oneidensis MR-1: impacts of cell surface appendage and polymer concentration

In wastewater treatment plants, optimizing bacterial flocculation and bacterial sludge dewatering requires a detailed understanding of the concomitant biological and physico-chemical processes governing the action of flocculating agent on living cells. Here we investigate the interactions between polyethyleneimine (PEI, 60,000 g/mol) and Shewanella oneidensis MR-1 lacking or not the lipopolysaccharide (LPS) O-antigen surface structure. Flocculation tests were performed on bacteria with/without LPS O-antigen after being exposed to 0-100 mg/L PEI concentrations. Measurements of electrophoretic mobility and bacterial aggregates size were complemented by transmission electron micrographs and atomic force microscopy images. While low PEI concentrations (<20 mg/L) lead to flocculation of both bare and LPS O-antigen-decorated bacterial strains, the lysis of bacterial membranes occurred at larger polymer concentrations for the latter, which highlights the protective role of LPS O-antigen against harmful PEI-mediated membrane alterations. Depending on polymer concentration, two types of bacterial aggregates are identified: one that solely integrates bacterial cells, and another that includes both cells and cell residues resulting from lysis (membrane and/or LPS fragments, and inner cell content materials). The latter is expected to significantly contribute to water entrapping in sludge and thus lower dewatering process efficiency.     

Reduced Photoinhibition under Low Irradiance Enhanced Kacip Fatimah (Labisia pumila Benth) Secondary Metabolites, Phenyl Alanine Lyase and Antioxidant Activity

A randomized complete block design experiment was designed to characterize the relationship between production of total flavonoids and phenolics, anthocyanin, photosynthesis, maximum efficiency of photosystem II (Fv/Fm), electron transfer rate (Fm/Fo), phenyl alanine lyase activity (PAL) and antioxidant (DPPH) in Labisia pumila var. alata, under four levels of irradiance (225, 500, 625 and 900 μmol/m(2)/s) for 16 weeks. As irradiance levels increased from 225 to 900 μmol/m(2)/s, the production of plant secondary metabolites (total flavonoids, phenolics and antocyanin) was found to decrease steadily. Production of total flavonoids and phenolics reached their peaks under 225 followed by 500, 625 and 900 μmol/m(2)/s irradiances. Significant positive correlation of production of total phenolics, flavonoids and antocyanin content with Fv/Fm, Fm/Fo and photosynthesis indicated up-regulation of carbon-based secondary metabolites (CBSM) under reduced photoinhibition on the under low light levels condition. At the lowest irradiance levels, Labisia pumila extracts also exhibited a significantly higher antioxidant activity (DPPH) than under high irradiance. The improved antioxidative activity under low light levels might be due to high availability of total flavonoids, phenolics and anthocyanin content in the plant extract. It was also found that an increase in the production of CBSM was due to high PAL activity under low light, probably signifying more availability of phenylalanine (Phe) under this condition.