Development and Characterization of Novel Biopolymer Derived from Abelmoschus esculentus L. Extract and Its Antidiabetic Potential.

Abelmoschus esculentus (Okra) is an important vegetable crop, widely cultivated around the world due to its high nutritional significance along with several health benefits. Different parts of okra including its mucilage have been currently studied for its role in various therapeutic applications. Therefore, we aimed to develop and characterize the okra mucilage biopolymer (OMB) for its physicochemical properties as well as to evaluate its in vitro antidiabetic activity. The characterization of OMB using Fourier-transform infrared spectroscopy (FT-IR) revealed that okra mucilage containing polysaccharides lies in the bandwidth of 3279 and 1030 cm-1, which constitutes the fingerprint region of the spectrum. In addition, physicochemical parameters such as percentage yield, percentage solubility, and swelling index were found to be 2.66%, 96.9%, and 5, respectively. A mineral analysis of newly developed biopolymers showed a substantial amount of calcium (412 mg/100 g), potassium (418 mg/100 g), phosphorus (60 mg/100 g), iron (47 mg/100 g), zinc (16 mg/100 g), and sodium (9 mg/100 g). The significant antidiabetic potential of OMB was demonstrated using α-amylase and α-glucosidase enzyme inhibitory assay. Further investigations are required to explore the newly developed biopolymer for its toxicity, efficacy, and its possible utilization in food, nutraceutical, as well as pharmaceutical industries.

Preliminary study to characterize plastic polymers using elemental analyser/isotope ratio mass spectrometry (EA/IRMS).

Plastic waste is a growing global environmental problem, particularly in the marine ecosystems, in consideration of its persistence. The monitoring of the plastic waste has become a global issue, as reported by several surveillance guidelines proposed by Regional Sea Conventions (OSPAR, UNEP) and appointed by the EU Marine Strategy Framework Directive. Policy responses to plastic waste vary at many levels, ranging from beach clean-up to bans on the commercialization of plastic bags and to Regional Plans for waste management and recycling. Moreover, in recent years, the production of plant-derived biodegradable plastic polymers has assumed increasing importance. This study reports the first preliminary characterization of carbon stable isotopes (δ13C) of different plastic polymers (petroleum- and plant-derived) in order to increase the dataset of isotopic values as a tool for further investigation in different fields of polymers research as well as in the marine environment surveillance. The δ13C values determined in different packaging for food uses reflect the plant origin of "BIO" materials, whereas the recycled plastic materials displayed a δ13C signatures between plant- and petroleum-derived polymers source. In a preliminary estimation, the different colours of plastic did not affect the variability of δ13C values, whereas the abiotic and biotic degradation processes that occurred in the plastic materials collected on beaches and in seawater, showed less negative δ13C values. A preliminary experimental field test confirmed these results. The advantages offered by isotope ratio mass spectrometry with respect to other analytical methods used to characterize the composition of plastic polymers are: high sensitivity, small amount of material required, rapidity of analysis, low cost and no limitation in black/dark samples compared with spectroscopic analysis.

Species and distribution of inorganic and organic phosphorus in enhanced phosphorus removal aerobic granular sludge.

The species and distribution of phosphorus (P) in an enhanced biological phosphorus removal (EBPR)-aerobic granular sludge (AGS) were fractionated and further analyzed. Results showed that microbial cells, extracellular polymeric substances (EPS) and mineral precipitates contributed about 73.7%, 17.6% and 5.3-6.4% to the total P (TP) of EBPR-AGS, respectively. Inorganic P (IP) species were orthophosphate, pyrophosphate and polyphosphate among which polyphosphate was the major P species in the AGS, cells and EPS. Monoester and diester phosphates were identified as the organic P (OP) species in the AGS and cells. Hydroxyapatite (Ca5(PO4)3OH) and calcium phosphate (Ca2(PO4)3) were the dominant P minerals accumulated in the core of the granules. Cells along with polyphosphate were mainly in the outer layer of AGS while EPS were distributed in the whole granules. Based on the above results, the distribution of IP and OP species in AGS has been conceived.

Effect of algae growth on aerobic granulation and nutrients removal from synthetic wastewater by using sequencing batch reactors.

The effect of algae growth on aerobic granulation and nutrients removal was studied in two identical sequencing batch reactors (SBRs). Sunlight exposure promoted the growth of algae in the SBR (Rs), forming an algal-bacterial symbiosis in aerobic granules. Compared to the control SBR (Rc), Rs had a slower granulation process with granules of loose structure and smaller particle size. Moreover, the specific oxygen uptake rate was significantly decreased for the granules from Rs with secretion of 25.7% and 22.5% less proteins and polysaccharides respectively in the extracellular polymeric substances. Although little impact was observed on chemical oxygen demand (COD) removal, algal-bacterial symbiosis deteriorated N and P removals, about 40.7-45.4% of total N and 44% of total P in Rs in contrast to 52.9-58.3% of TN and 90% of TP in Rc, respectively. In addition, the growth of algae altered the microbial community in Rs, especially unfavorable for Nitrospiraceae and Nitrosomonadaceae.

Effects of salinity build-up on biomass characteristics and trace organic chemical removal: implications on the development of high retention membrane bioreactors.

This study investigated the impact of salinity build-up on the performance of membrane bioreactor (MBR), specifically in terms of the removal and fate of trace organic chemicals (TrOCs), nutrient removal, and biomass characteristics. Stepwise increase of the influent salinity, simulating salinity build-up in high retention MBRs, adversely affected the metabolic activity in the bioreactor, thereby reducing organic and nutrient removal. The removal of hydrophilic TrOCs by MBR decreased due to salinity build-up. By contrast, with the exception of 17α-ethynylestradiol, the removal of all hydrophobic TrOCs was not affected at high salinity. Moreover, salinity build-up had negligible impact on the residual accumulation of TrOCs in the sludge phase except for a few hydrophilic compounds. Additionally, the response of the biomass to salinity stress also dramatically enhanced the release of both soluble microbial products (SMP) and extracellular polymeric substances (EPS), leading to severe membrane fouling.

Nutrient release, recovery and removal from waste sludge of a biological nutrient removal system.

The uncontrolled release of nutrients from waste sludge results in nitrogen and phosphorus overloading in wastewater treatment plants when supernatant is returned to the inlet. A controlled release, recovery and removal of nutrient from the waste sludge of a Biological Nutrient Removal system (BNR) are investigated. Results showed that the supernatant was of high mineral salt, high electrical conductivity and poor biodegradability, in addition to high nitrogen and phosphorus concentrations after the waste sludge was hydrolysed through sodium dodecyl sulphate addition. Subsequently, over 91.8% of phosphorus and 10.5% of nitrogen in the supernatants were extracted by the crystallization method under the conditions of 9.5 pH and 400 rpm. The precipitate was mainly struvite according to X-ray diffraction and morphological examination. A multistage anoxic-oxic Moving Bed Biofilm Reactor (MBBR) was then adopted to remove the residual carbon, nitrogen and phosphorus in the supernatant. The MBBR exhibited good performance in simultaneously removing carbon, nitrogen and phosphorus under a short aeration time, which accounted for 31.25% of a cycle. Fluorescence in situ hybridization analysis demonstrated that nitrifiers presented mainly in floc, although higher extracellular polymeric substance content, especially DNA, appeared in the biofilm. Thus, a combination of hydrolysis and precipitation, followed by the MBBR, can complete the nutrient release from the waste sludge of a BNR system, recovers nutrients from the hydrolysed liquor and removes nutrients from leftovers effectively.

Involvement of GPI-anchored lipid transfer proteins in the development of seed coats and pollen in Arabidopsis thaliana.

The non-specific lipid transfer proteins (nsLTPs) constitute a large protein family specific for plants. Proteins from the family are found in all land plants but have not been identified in green algae. Their in vivo functions are still disputed although evidence is accumulating for a role of these proteins in cuticle development. In a previous study, we performed a co-expression analysis of glycosylphosphatidylinositol (GPI)-anchored nsLTPs (LTPGs), which suggested that these proteins are also involved in the accumulation of suberin and sporopollenin. Here, we follow up the previous co-expression study by characterizing the phenotypes of Arabidopsis thaliana lines with insertions in LTPG genes. The observed phenotypes include an inability to limit tetrazolium salt uptake in seeds, development of hair-like structures on seeds, altered pollen morphologies and decreased levels of ω-hydroxy fatty acids in seed coats. The observed phenotypes give further support for a role in suberin and sporopollenin biosynthesis or deposition in A. thaliana.

Nanometer-scale visualization and structural analysis of the inorganic/organic hybrid structure of Gallionella ferruginea twisted stalks.

The so-called Fe/Mn-oxidizing bacteria have long been recognized for their potential to form extracellular iron hydroxide or manganese oxide structures in aquatic environments. Bacterial species belonging to the genus Gallionella, one type of such bacteria, oxidize iron and produce uniquely twisted extracellular stalks consisting of iron oxide-encrusted inorganic/organic fibers. This paper describes the ultrastructure of Gallionella cells and stalks and the visualized structural and spatial localization of constitutive elements within the stalks. Electron microscopy with energy-dispersive X-ray microanalysis showed the export site of the stalk fibers from the cell and the uniform distribution of iron, silicon, and phosphorous in the stalks. Electron energy-loss spectroscopy revealed that the stalk fibers had a central carbon core of bacterial exopolymers and that aquatic iron interacted with oxygen at the surface of the carbon core, resulting in deposition of iron oxides at the surface. This new knowledge of the structural and spatial associations of iron with oxygen and carbon provides deeper insights into the unique inorganic/organic hybrid structure of the stalks.

Magnetic modulation biosensing for rapid and homogeneous detection of biological targets at low concentrations.

This paper reviews the development of a magnetic modulation biosensing (MMB) system for rapid, simple and sensitive detection of biological targets in homogeneous solution at low concentrations. It relies on condensation and modulation of the fluorescent-labeled probes attached to magnetic beads using an alternating magnetic field gradient. Condensation of the beads from the entire volume increases the signal while modulation separates the signal from the background noise of the non-magnetized solution. We first discuss the motivation and challenges in specific DNA sequences detection as well as current approaches to overcome some of these challenges. We then present the MMB system, DNA detection schemes and magnetic beads manipulation in solution. Rapid detection at sub-picomolar concentrations of fluorescent-labeled probes as well as of coding sequences of the non-structural Ibaraki virus protein 3 (NS3) complementary DNA (cDNA) without any washing or separation step is also reviewed. Finally, we show preliminary results of protein detection using a 'sandwich'-based assay.

Off-line comprehensive 2-dimensional hydrophilic interaction x reversed phase liquid chromatography analysis of procyanidins.

The development of an off-line comprehensive 2-dimensional liquid chromatography (2-D-LC) method for the analysis of procyanidins is reported. In the first dimension, oligomeric procyanidins were separated according to molecular weight by hydrophilic interaction chromatography (HILIC), while reversed phase LC was employed in the second dimension to separate oligomers based on hydrophobicity. Fluorescence, UV and electrospray ionisation mass spectrometry (ESI-MS) were employed for identification purposes. The combination of these orthogonal separation methods is shown to represent a significant improvement compared to 1-dimensional methods for the analysis of complex high molecular weight procyanidin fractions, by simultaneously providing isomeric and molecular weight information. The low correlation (r(2)<0.2100) between the two LC modes afforded a practical peak capacity in excess of 2300 for the optimal off-line method. The applicability of the method is demonstrated for the analysis of phenolic extracts of apple and cocoa.

Two-dimensional DOSY experiment with Excitation Sculpting water suppression for the analysis of natural and biological media.

The Bipolar Pulse Pair Stimulated Echo NMR pulse sequence was modified to blend the original Excitation Sculpting water signal suppression. The sequence is a powerful tool to generate rapidly, with a good spectrum quality, bidimensional DOSY experiments without solvent signal, thus allowing the analysis of complex mixtures such as plant extracts or biofluids. The sequence has also been successfully implemented for a protein at very-low concentration in interaction with a small ligand, namely the salivary IB5 protein binding the polyphenol epigallocatechine gallate. The artifacts created by this sequence can be observed, checked and removed thanks to NPK and NMRnotebook softwares to give a perfect bidimensional DOSY spectrum.

Ecological effects of aquaculture on living and non-living suspended fractions of the water column: a meta-analysis.

The effects of aquaculture on the ecology of the water column have been extensively studied in the last two decades. However, to date, it has not been possible to extrapolate homogeneous information from the peer-reviewed literature. In the present study, 68 peer-reviewed articles were analysed and about 1087 study cases were used to test whether worldwide cultivations of aquatic organisms (shrimps, fish, bivalves and polyculture) have a differential effect on living and non-living fractions of the water column (suspended matter, chlorophyll-a, particulate organic carbon, nitrogen and phosphorus, bacteria and plankton). Meta-analysis feasibility depends on obtaining an estimate of the effect size from every study, and the most common measure of effect size (Hedges'd) is the difference between means of controls and impacts standardised by dividing by the pooled standard deviation. Shrimp, fish and bivalve cultivation differentially affected water column dynamics, with a general major impact on bacteria and phytoplankton. In addition, results showed that the water column dynamics are probably affected by organic aquaculture loading but, due to the substantial heterogeneity across studies, the information available on the effects can be considered partially flawed and therefore not sufficient to either support or exclude the notion that different forms of aquaculture affect ecological processes of the water column.

Hypoglycemic effects of Ganoderma applanatum and Collybia confluens exo-polymers in streptozotocin-induced diabetic rats.

The hypoglycemic effects of Ganoderma applanatum exo-polymer (GAE) and Collybia confluens exo-polymer (CCE) produced by submerged mycelial cultures in streptozotocin (STZ)-induced diabetic rats were investigated. Hypoglycemic effects were achieved in both the GAE- and CCE-treated groups by administration at a level of 100 mg/kg body weight (BW) daily for 3 weeks. The administration of GAE and CCE substantially reduced the plasma glucose levels by as much as 22.0% and 25.9%, respectively, when compared with the control group. The GAE and CCE also lowered the plasma total cholesterol and triglyceride levels by 20.3% and 22.5%, and by 22.7% and 25.5%, respectively. Furthermore, the activity of alanine transaminase (ALT) and aspartate transaminase (AST) was decreased by 23.2% and 20.7% in the GAE-treated group, and it was also reduced by 28.7% and 23.6% in the CCE-treated group. The results strongly demonstrate the potential of GAE and CCE in combating diabetes in experimental animals.

Role of extracellular exopolymers in biological phosphorus removal.

Three sequencing batch reactors (SBRs) supplied with different carbon sources were investigated. The system supplied with glucose gained the best enhanced biological phosphorus removal (EBPR), although all of them were seeded from the same sludge. With the measurement of poly-beta-hydroxyalkanoate (PHA) concentration, phosphorus content in sludge and extracellular exopolymers (EPs) with scanning electron microscopy (SEM) combined with energy dispersive spectrometry (EDS), it was found that the biosorption effect of EPs played an important role in phosphorus removal and that the amount of PHA at the end of anaerobic phase was not the only key factor to determine the following phosphorus removal efficiency.

Cytochemistry and immunolocalisation of polysaccharides and proteoglycans in the endosperm of green Arabica coffee beans.

The major noncellulosic polysaccharides and proteoglycans in the coffee bean (Coffea arabica) cell wall are (galacto)mannans and arabinogalactan proteins. Immunological and chemical probes demonstrated that the mannans and arabinogalactan proteins were located continuously across the width of the cell wall, but that the concentration of different structural epitopes within these polysaccharide types showed considerable spatial variation. For the mannans this was implied by the striated pattern demonstrated by fluctuation of the affinity between the mannan monoclonal antibody BGM C6 and (galacto)mannan. The arabinogalactan proteins labelled by the Yariv reagent and the arabinogalactan protein-specific antibody LM2 appeared to be located in all regions of the wall except the middle lamella, but showed some differences in intensity of labelling. However, the LM6 antibody, specific for (1-->5)-alpha-arabinan epitopes, was located only as a compact region adjacent to the cell lumen in the body of the endosperm; though, it did label throughout the wall of epidermal cells. This implied that either some of the more highly arabinosylated arabinogalactan proteins contained contiguous 5-arabinosyl residues or that a rhamnogalacturonan which contained 5-arabinosyl residues as side chains existed in the cell wall. In either case the polymers were very restricted in their distribution. A second category of pectin, a homogalacturonan detected by JIM7, was located only in the middle lamella region. The architecture of the wall, as revealed by resin etching, appeared to reflect the chemical heterogeneity, with three distinct physical zones identifiable in a cross section across a single wall.

Polyubiquitin in crustacean striated muscle: increased expression and conjugation during molt-induced claw muscle atrophy.

The claw muscles of decapod crustaceans undergo a molt-induced atrophy to facilitate withdrawal of the claws at ecdysis. Polyubiquitin expression, as well as the levels of ubiquitin conjugates, a ubiquitin-conjugating enzyme involved in the ATP/ubiquitin-dependent proteolytic pathway (crustacean E2(16 kDa) homolog of Drosophila UbcD1), and proteasome, were examined to determine the role of ATP/ubiquitin-dependent proteolysis in the enhanced degradation of myofibrillar proteins during muscle atrophy. A partial-length clone (1.7 kb) of polyubiquitin was isolated from a lobster muscle cDNA library; the 5' end lacked the 5' untranslated region (UTR) and the beginning of the first ubiquitin monomer, while the 3' end contained the terminal ubiquitin monomer and 3' UTR. The deduced amino acid sequence was 100% identical with that from Manduca, Drosophila, and human. In land crab claw muscle, the polyubiquitin mRNA (2.7 kb) increased about 5-fold and ubiquitin-protein conjugates (> 200 kDa) increased about 8-fold during atrophy. In contrast, the level of a ubiquitin-conjugating enzyme (E2(16 kDa)) remained unchanged. The proteasome, which constitutes the catalytic core of the ATP/ubiquitin-dependent proteinase complex, increased about 2-fold during proecdysis, reaching its highest level immediately before ecdysis. These results suggest that the ATP/ubiquitin-dependent proteolytic pathway contributes to the changes in protein metabolism that occur during molt-induced muscle atrophy.