Search results for: protein oxidation

Authors: Abu Salim Mustafa

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Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: mycobacterium tuberculosis, PPE68, peptides, vaccine

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Authors: Xing Lv, Hui-Qin Xu

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The present study aimed to reveal the mechanism in aggravating STZ induced diabetic nephropathy (DN) by AGEs (advanced glycation end products). At the eighth day, 20 diabetic mice were randomly divided into STZ group and combination (combine AGEs with STZ) group. Simultaneously, AGEs group and normal group were set. Only mice in AGEs group, combination group were fed with high-AGEs diets. Mice diabetic conventional indicators, biochemical analysis were measured. Among the indictors, food consumptions, water intake, urine output, blood glucose, urine protein, urine creatinine, serum urea nitrogen were increased significantly in STZ, combination groups. The AGEs levels in combination group increased significantly when compared with STZ group. Weights and insulin levels in the STZ, combination groups were decreased significantly when compared with normal group, and the difference was significantly between AGEs group and STZ group. As a conclusion, AGEs play an important role in the DN development, inducing kidney damages.

Keywords: AGEs, diabetic nephropathy, serum urea nitrogen, urine protein

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Authors: Hongquan Zhang

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Inspired by endogenous protein motors, researchers have constructed various synthetic DNA motors based on the specificity and predictability of Watson-Crick base pairing. However, the application of DNA motors to signal amplification and biosensing is limited because of low mobility and difficulty in real-time monitoring of the walking process. The objective of our work was to construct a new type of DNA motor termed target-triggered DNA motors that can walk for hundreds of steps in response to a single target binding event. To improve the mobility and processivity of DNA motors, we used gold nanoparticles (AuNPs) as scaffolds to build high-density, three-dimensional tracks. Hundreds of track strands are conjugated to a single AuNP. To enable DNA motors to respond to specific protein and nucleic acid targets, we adapted the binding-induced DNA assembly into the design of the target-triggered DNA motors. In response to the binding of specific target molecules, DNA motors are activated to autonomously walk along AuNP, which is powered by a nicking endonuclease or DNAzyme-catalyzed cleavage of track strands. Each moving step restores the fluorescence of a dye molecule, enabling monitoring of the operation of DNA motors in real time. The motors can translate a single binding event into the generation of hundreds of oligonucleotides from a single nanoparticle. The motors have been applied to amplify the detection of proteins and nucleic acids in test tubes and live cells. The motors were able to detect low pM concentrations of specific protein and nucleic acid targets in homogeneous solutions without the need for separation. Target-triggered DNA motors are significant for broadening applications of DNA motors to molecular sensing, cell imagining, molecular interaction monitoring, and controlled delivery and release of therapeutics.

Keywords: biosensing, DNA motors, gold nanoparticles, signal amplification

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Authors: Valentina Paolucci, Jessica De Santis, Vittorio Ricci, Giacomo Giorgi, Carlo Cantalini

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Despite their potential in gas sensing applications, the major drawback of 2D exfoliated metal dichalcogenides (MDs) is that they suffer from spontaneous oxidation in air, showing poor chemical stability under dry/wet conditions even at room temperature, limiting their practical exploitation. The aim of this work is to validate a synthesis strategy allowing microstructural and electrical stabilization of the oxides that inevitably form on the surface of 2D dichalcogenides. Taking advantage of spontaneous oxidation of MDs in air, we report on liquid phase exfoliated 2D-SnSe2 flakes annealed in static air at a temperature below the crystallization temperature of the native a-SnO2 oxide. This process yields a new class of 2D Layered Amorphous Metal Oxides Sensors (LAMOS), specifically few-layered amorphous a-SnO2, showing excellent gas sensing properties. Sensing tests were carried out at low operating temperature (i.e. 100°C) by exposing a-SnO2 to both oxidizing and reducing gases (i.e. NO2, H2S and H2) and different relative humidities ranging from 40% to 80% RH. The formation of stable nanosheets of amorphous a-SnO2 guarantees excellent reproducibility and stability of the response over one year. These results pave the way to new interesting research perspectives out considering the opportunity to synthesize homogeneous amorphous textures with no grain boundaries, no grains, no crystalline planes with different orientations, etc., following gas sensing mechanisms that likely differ from that of traditional crystalline metal oxide sensors. Moreover, the controlled annealing process could likely be extended to a large variety of Transition Metal Dichalcogenides (TMDs) and Metal Chalcogenides (MCs), where sulfur, selenium, or tellurium atoms can be easily displaced by O2 atoms (ΔG < 0), enabling the synthesis of a new family of amorphous interfaces.

Keywords: layered 2D materials, exfoliation, lamos, amorphous metal oxide sensors

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Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

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G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

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Authors: Rinat Arbel-Goren, Joel Stavans

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Cell-to-cell variations in transcript or protein abundance, called noise, may give rise to phenotypic variability between isogenic cells, enhancing the probability of survival under stress conditions. These variations may be introduced by post-transcriptional regulatory processes such as non-coding, small RNAs stoichiometric degradation of target transcripts in bacteria. We study the iron homeostasis network in Escherichia coli, in which the RyhB small RNA regulates the expression of various targets as a model system. Using fluorescence reporter genes to detect protein levels and single-molecule fluorescence in situ hybridization to monitor transcripts levels in individual cells, allows us to compare noise at both transcript and protein levels. The experimental results and computer simulations show that extrinsic noise buffers through a feed-forward loop configuration the increase in variability introduced at the transcript level by iron deprivation, illuminating the important role that extrinsic noise plays during stress. Surprisingly, extrinsic noise also decouples of fluctuations of two different targets, in spite of RyhB being a common upstream factor degrading both. Thus, phenotypic variability increases under stress conditions by the decoupling of target fluctuations in the same cell rather than by increasing the noise of each. We also present preliminary results on the adaptation of cells to prolonged iron deprivation in order to shed light on the evolutionary role of post-transcriptional downregulation by small RNAs.

Keywords: cell-to-cell variability, Escherichia coli, noise, single-molecule fluorescence in situ hybridization (smFISH), transcript

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Authors: Sireethorn Nimitvilai, Ussanee Poolvivatchaikarn, Nuchanart Tomeun

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Objective: To determine the simple laboratory parameters to predict disease severity among COVID-19 patients in resource-limited settings. Material and methods: A retrospective cohort study was conducted at Nakhonpathom Hospital, a 722-bed tertiary care hospital, with an average of 50,000 admissions per year, during April 15 and May 15, 2021. Eligible patients were adults aged ≥ 15 years who were hospitalized with COVID-19. Baseline characteristics, comorbid conditions ad laboratory findings at admission were collected. Predictive factors for severe COVID-19 infection were analyzed. Result: There were 207 patients (79 male and 128 female) and the mean age was 46.7 (16.8) years. Of these, 39 cases (18.8%) were severe and 168 (81.2%) cases were non-severe. Factors associated with severe COVID-19 were neutrophil to lymphocyte ratio ≥ 4 (OR 8.1, 95%CI 2.3-20.3, P < 0.001) and C-reactive protein to albumin ratio ≥ 10 (OR 3.49, 95%CI 1.3-9.1, p 0.01). Conclusions: Complete blood counts, C-reactive protein and albumin are simple, inexpensive, widely available tests and can be used to predict severe COVID-19 in resource-limited settings.

Keywords: COVID-19, predictor of severity, resource-limiting settings, simple laboratory parameters

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Authors: Mohammed Sabbah, Prospero Di Pierro, Raffaele Porta

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Protein-based edible films and coatings have attracted an increasing interest in recent years since they might be used to protect pharmaceuticals or improve the shelf life of different food products. Among them, several plant proteins represent an abundant, inexpensive and renewable raw source. These natural biopolymers are used as film forming agents, being able to form intermolecular linkages by various interactions. However, without the addition of a plasticizing agent, many biomaterials are brittle and, consequently, very difficult to be manipulated. Plasticizers are generally small and non-volatile organic additives used to increase film extensibility and reduce its crystallinity, brittleness and water vapor permeability. Plasticizers normally act by decreasing the intermolecular forces along the polymer chains, thus reducing the relative number of polymer-polymer contacts, producing a decrease in cohesion and tensile strength and thereby increasing film flexibility allowing its deformation without rupture. The most commonly studied plasticizers are polyols, like glycerol (GLY) and some mono or oligosaccharides. In particular, GLY not only increases film extensibility but also migrates inside the film network often causing the loss of desirable mechanical properties of the material. Therefore, replacing GLY with a different plasticizer might help to improve film characteristics allowing potential industrial applications. To improve film properties, it seemed of interest to test as plasticizers some cationic small molecules like polyamines (PAs). Putrescine, spermidine (SPD), and spermine are PAs widely distributed in nature and of particular interest for their biological activities that may have some beneficial health effects. Since PAs contains amino instead of hydroxyl functional groups, they are able to trigger ionic interactions with negatively charged proteins. Bitter vetch (Vicia ervilia; BV) is an ancient grain legume crop, originated in the Mediterranean region, which can be found today in many countries around the world. This annual Vicia genus shows several favorable features, being their seeds a cheap and abundant protein source. The main objectives of this study were to investigate the effect of different concentrations of SPD on the mechanical and permeability properties of films prepared with native or heat denatured BV proteins in the presence of different concentrations of SPD and/or GLY. Therefore, a BV seed protein concentrate (BVPC), containing about 77% proteins, was used to prepare film forming solutions (FFSs), whereas GLY and SPD were added as film plasticizers, either singly or in combination, at various concentrations. Since a primary plasticizer is generally defined as a molecule that when added to a material makes it softer, more flexible and easier to be processed, our findings lead to consider SPD as a possible primary plasticizer of protein-based films. In fact, the addition of millimolar concentrations of SPD to BVPC FFS allowed obtaining handleable biomaterials with improved properties. Moreover, SPD can be also considered as a secondary plasticizer, namely an 'extender', because of its ability even to enhance the plasticizing performance of GLY. In conclusion, our studies indicate that innovative edible protein-based films and coatings can be obtained by using PAs as new plasticizers.

Keywords: edible films, glycerol, plasticizers, polyamines, spermidine

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Authors: Shenghu Feng, Jun Cheng

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The molecular mechanism underlying nonalcoholic fatty liver disease (NAFLD) progression to hepatocellular carcinoma (HCC) remains unknown. In this study, immunohistochemistry staining result showed that NS5ABP37 protein expression decreased as with increasing degree of HCC malignancy. In agreement, NS5ABP37 protein overexpression significantly suppressed cell proliferation, caused G1/S cell cycle arrest, and induced apoptosis by increasing caspase-3/7 activity and cleaved caspase-3 levels. In addition, NS5ABP37 overexpression resulted in decreased intracellular TG and TC contents, with level reduction in SREBPs and downstream effectors. Furthermore, NS5ABP37 overexpression decreased SREBP1c and SREBP2 levels by inducing their respective promoters. Finally, ROS levels and ER-stress were both induced by NS5ABP37 overexpression. These findings together demonstrate that NS5ABP37 inhibits cancer cell proliferation and promotes apoptosis, by altering SREBP-dependent lipogenesis and cholesterogenesis in HepG2 cells and inducing oxidative stress and ER stress.

Keywords: NS5ABP37, liver cancer, lipid metabolism, oxidative stress, ER stress

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Authors: Sofia G. Meirinho, Luis G. Dias, António M. Peres, Lígia R. Rodrigues

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The emerging development of electrochemical aptasen sors has enabled the easy and fast detection of protein biomarkers in standard and real samples. Biomarkers are produced by body organs or tumours and provide a measure of antigens on cell surfaces. When detected in high amounts in blood, they can be suggestive of tumour activity. These biomarkers are more often used to evaluate treatment effects or to assess the potential for metastatic disease in patients with established disease. Osteopontin (OPN) is a protein found in all body fluids and constitutes a possible biomarker because its overexpression has been related with breast cancer evolution and metastasis. Currently, biomarkers are commonly used for the development of diagnostic methods, allowing the detection of the disease in its initial stages. A previously described RNA aptamer was used in the current work to develop a simple and sensitive electrochemical aptasensor with high affinity for human OPN. The RNA aptamer was biotinylated and immobilized on a gold electrode by avidin-biotin interaction. The electrochemical signal generated from the aptamer–target molecule interaction was monitored electrochemically using cyclic voltammetry in the presence of [Fe (CN) 6]−3/− as a redox probe. The signal observed showed a current decrease due to the binding of OPN. The preliminary results showed that this aptasensor enables the detection of OPN in standard solutions, showing good selectivity towards the target in the presence of others interfering proteins such as bovine OPN and bovine serum albumin. The results gathered in the current work suggest that the proposed electrochemical aptasensor is a simple and sensitive detection tool for human OPN and so, may have future applications in cancer disease monitoring.

Keywords: osteopontin, aptamer, aptasensor, screen-printed electrode, cyclic voltammetry

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Authors: Kai Pei Huang, Ting Chung Hung, Li Ching Wu

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Amyloidosis refers to a variety of conditions wherein amyloid proteins are abnormally deposited in organ or tissues and cause harm. Among the several forms of amyloidosis, the principal types of that in inpatient medical services are the AL amyloidosis (primary) and AA amyloidois (secondary). AL Amyloidois is due to deposition of protein derived from overproduction of immunoglobulin light chain in plasma cell myeloma. Furthermore, it is a systemic disorder that can present with a variety of symptoms, including heavy proteinemia and edema, heptosplenomegaly, otherwise unexplained heart failure. We reported a 78-year-old female presenting dysuria, oliguria and leg edema for several months. Laboratory data showed proteinuria (UPCR:1679.8), leukocytosis (WBC:16.2 x 10^3/uL), results of serum urea nitrogen (39mg/dL), creatinine (0.76 mg/dL), IgG (748 mg/dL.), IgA (635 mg/dL), IgM (63 mg/dL), kappa light chain(18.8 mg/dL), lambda light chain (110.0 mg/dL) and kappa/lambda ratio (0.17). Renal biopsy found amyloid fibrils in glomerular mesangial area, and Congo red stain highlights amyloid deposition in glomeruli. Additional lab studies included serum protein electrophoresis, which shows a major monoclonal peak in β region and minor small peak in gamma region, and the immunotyping studies for serum showed two IgA/λ type. We treated sample with beta-mercaptoethanol which reducing the polymerized immunoglobulin to clarify two IgA/λ are secreted from the same plasma cell clone in bone marrow. Later examination confirmed it existed plasma cell infiltration in bone marrow, and the immunohistochemical staining showed monotypic for λ light chain and are positive for IgA. All findings mentioned above reveal it is a case of plasma cell myeloma with λ Light Chain Amyloidosis.

Keywords: amyloidosis, immunoglobulin light chain, plasma cell myeloma, serum protein electrophoresis

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Authors: Meskure Pak, Nihal Buyukuslu

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During Ramadan Muslims refrain from eating and drinking from dawn to sunset. Transformation of the eating habits cause profound changes in body composition. This study was performed during Ramadan of June-July 2015 with the regular fasting healthy adults (15 women and 15 men). The participants who were not fasting the whole month, have chronic diseases, pregnant and lactated were excluded. All attendances were informed about study. Written informed consent was taken from the voluntary participants. The work was approved by the Ethics and Research Committee of Istanbul Medipol University, Turkey. A questionnaire was conducted to determine the nutritional status, demographic and anthropometric data at the beginning, in the middle and at the end of Ramadan. Statistical Package for Social Sciences version 18.0 (SPSS, Chicago, IL, USA) was used for analyses. The mean ages of women and men were 34.4±9.45 and 28.9±10.55 years respectively. The BMI values (kg/m2) were slightly increased in men (26.5±3.9 to 26.2±3.7) and decreased in women (22.5±3.5 to 23.3±4.5). However the differences in BMI values between genders were not significant. Waist circumferences (WC) (cm) decreased in both women (80.2±14.6 to 79.4±17.7) and men (98.9±8.4 to 95.2±11.0) through the Ramadan. Fat percentages of women (27.0±71) increased in the middle of Ramadan (28.4±7.8) and decreased at the end of Ramadan (27.8±8.3). The fat percentages of men (21.5±6.3) were not affected in the middle of Ramadan (21.5±6.4) however decreased at the end of Ramadan (20.8±6.2). The total change in fat mass from beginning to end of Ramadan was higher in women than in men. The daily energy intake was higher in men than in women during Ramadan. In the middle of Ramadan energy intake (kcal) was reached to the highest level (2057.8±693.1) and at the end of Ramadan it decreased to the beginning level (1656.7±553.2) for men. However, daily energy intake of women slightly decreased from the beginning (1410.0±359.7) to the end (1409.2±366.7) of Ramadan. The comparison of energy intake between men and women was significant in the middle of Ramadan (p < 0.05). Water consumptions for both groups were increased in Ramadan fasting period. In comparison with the beginning of Ramadan, daily carbohydrate and fat consumptions increased and the consumption of protein decreased for men and for women at the end of Ramadan. The gender comparison resulted in a significant increase for protein and carbohydrate consumption of men in the middle of Ramadan (p < 0.05). In the first two weeks, the daily energy intake, the consumption of carbohydrates and fats seemed to increase for both men and women. However the later days of Ramadan daily fat consumption decreased to the level of beginning consumption levels which may indicate the nutritional adaptation period. In spite of the consumption of protein sources such as meat, poultry and egg increased, the decrease in the total amount of protein consumed in Ramadan may be due to a decrease in the consumption of milk and dairy products. In conclusion, the nutritional habits and preferred foods changed during Ramadan as a result affected the body composition.

Keywords: body composition, fasting, nutritional status, Ramadan

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Authors: P. Muñiz, R. Del Pino-García , M.D. Rivero-Pérez, J. García-Lomillo, M. L. González-SanJosé

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Grape, wine and wine pomace could improve the antioxidant status in the vasculature in terms of plasma antioxidant capacity and oxidation biomarkers, partly due to their high content in polyphenols. The current study aimed to evaluate the protection of a powdered product obtained from wine pomace (WPP) against oxidative damage associated to diabetes. Streptozotocin-induced diabetic (STZ) male Wistar rats and non-diabetic control (C) rats initially weighting 300±10 mg were supplemented with 100 mg of WPP or vehicle for 4 weeks. Blood glucose levels and body weight (BW) were measured weekly. Total antioxidant capacity (TAC) assessed using the ABTS method, and F2α-Isoprostanes (F2-IsoPs) quantified by GC-MS were measured in plasma collected at the end of this experiment. Blood glucose levels tended to increase in the STZ group along the study. Supplementation maintained relatively stable during the whole experiment the blood glucose values in STZ+WPP rats. A weight loss of BW in STZ rats respect to C rats was observed after 4 weeks, whereas the decrease in BW of STZ+WPP group showed a tendency to improve at the end of the study. TAC values significantly decreased around 11% only in plasma of STZ rats. The rest of groups showed plasma TAC values about 8 mM Trolox. Increased levels of F2-IsoPs (around 25%) were also observed in plasma of STZ rats compared to the supplemented rats, revealing a protective effect of WPP against lipid peroxidation. In conclusion, 4-week supplementation with a product derived from winery by-products improved weight loss, plasma TAC, and lipid oxidation biomarkers in Type I diabetic rats.

Keywords: blood glucose, grape polyphenols, F2α-isoprostanes, type I diabetes, oxidative stress

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Authors: Phakamani Xaba, Robert Huberts, Bilainu Oboirien

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The present study was aimed at developing & comparing two neural-network-based predictive models to predict shelf-life/product age of South African margarine using free fatty acid (FFA), water droplet size (D3.3), water droplet distribution (e-sigma), moisture content, peroxide value (PV), anisidine valve (AnV) and total oxidation (totox) value as input variables to the model. Brick margarine products which had varying ages ranging from fresh i.e. week 0 to week 47 were sourced. The brick margarine products which had been stored at 10 & 25 °C and were characterized. JMP and MATLAB models to predict shelf-life/ margarine age were developed and their performances were compared. The key performance indicators to evaluate the model performances were correlation coefficient (CC), root mean square error (RMSE), and mean absolute percentage error (MAPE) relative to the actual data. The MATLAB-developed model showed a better performance in all three performance indicators. The correlation coefficient of the MATLAB model was 99.86% versus 99.74% for the JMP model, the RMSE was 0.720 compared to 1.005 and the MAPE was 7.4% compared to 8.571%. The MATLAB model was selected to be the most accurate, and then, the number of hidden neurons/ nodes was optimized to develop a single predictive model. The optimized MATLAB with 10 neurons showed a better performance compared to the models with 1 & 5 hidden neurons. The developed models can be used by margarine manufacturers, food research institutions, researchers etc, to predict shelf-life/ margarine product age, optimize addition of antioxidants, extend shelf-life of products and proactively troubleshoot for problems related to changes which have an impact on shelf-life of margarine without conducting expensive trials.

Keywords: margarine shelf-life, predictive modelling, neural networks, oil oxidation

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Authors: Vishnu V. Pillai, Sunil P. Lonkar, Akhil M. Abraham, Saeed M. Alhassan

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An effectual technology for wastewater treatment is a great demand now in order to encounter the water pollution caused by organic pollutants. Photocatalytic oxidation technology is widely used in removal of such unsafe contaminants. Among the semi-conducting metal oxides, robust and thermally stable TiO2 has emerged as a fascinating material for photocatalysis. Enhanced catalytic activity was observed for nanostructured TiO2 due to its higher surface, chemical stability and higher oxidation ability. However, higher charge carrier recombination and wide band gap of TiO2 limits its use as a photocatalyst in the UV region. It is desirable to develop a photocatalyst that can efficiently absorb the visible light, which occupies the main part of the solar spectrum. Hence, in order to extend its photocatalytic efficiency under visible light, TiO2 nanoparticles are often doped with metallic or non-metallic elements. Non-metallic doping of TiO2 has attracted much attention due to the low thermal stability and enhanced recombination of charge carriers endowed by metallic doping of TiO2. Amongst, sulfur doped TiO2 is most widely used photocatalyst in environmental purification. However, the most of S-TiO2 synthesis technique uses toxic chemicals and complex procedures. Hence, a facile, scalable and environmentally benign preparation process for S-TiO2 is highly desirable. In present work, we have demonstrated new and facile solid-state reaction method for S-TiO2 synthesis that uses abundant elemental sulfur as S source and moderate temperatures. The resulting nano-sized S-TiO2 has been successfully employed as visible light photocatalyst in methylene blue dye removal from aqueous media.

Keywords: ecofriendly, nanomaterials, methylene blue, photocatalysts

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Authors: V. Karuppaiah, J. C. Padaria, C. Srivastava

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The tobacco caterpillar, Spodoptera litura Fab (Lepidoptera: Noctuidae) is a polyphagous pest various field and horticulture crops in India. Pest had virtually developed resistance to all commonly used insecticides. Enhanced detoxification is the prime mechanism that is dictated by detoxification different enzymes and carboxylesterase is one of the major enzyme responsible development of resistance. In India, insecticide resistance studies on S. litura are mainly deployed on detoxification enzymes activity and investigation at gene level alteration i.e. at nucleotide level is very merger. In the present study, we collected the S. litura larvae from three different cauliflower growing belt viz., IARI, New Delhi (Delhi), Palari, Sonepat (Haryana) and Varanasi (Uttar Pradesh) to study the role of carboxylesterase activity and its gene level variation The CarE activity was measured using UV-VIS spectrophotometer with 3rd instar larvae of S. litura. The elevated activity of CarE was observed in Sonepat strain (28.09 ± 0.09 µmol/min/mg of protein) followed by Delhi (26.72 ± 0.04 µmol/min/mg of protein) and Varanasi strain (10.00 ± 0.44 µmol/min/mg of protein) of S. litura. The genomic DNA was isolated from 3rd instar larvae and CarE gene was amplified using a primer sequence, F:5’tccagagttccttgtcaggcac3’; R:5’ctgcatcaagcatgtctc3. CarE gene, about 500bp was partially amplified, sequenced and submitted to NCBI (Accession No. KF835886, KF835887 and KF835888). The sequence data revealed polymorphism at nucleotide level in all the three strains and gene found to have 88 to 97% similarity with previous available nucleotide sequences of S. litura, S. littoralis and S. exiqua. The polymorphism at the nucleotide level could be a reason for differential activity of carboxylesterase enzymes among the strains. However, investigation at gene expression level would be useful to analyze the overproduction of carboxylesterase enzyme.

Keywords: carboxylesterase, CarE gene, nucleotide polymorphism, insecticide resistance, spodoptera litura

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Authors: Ana M. T. Mata, Alexiane Ligneul

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Textile effluents are usually heavily loaded with organic carbon and color compounds, the latter being azo dyes in an estimated 70% of the case effluent posing a major challenge in environmental protection. In this study, the ecotoxicity of simulated textile effluent after biological treatment with anaerobic and aerobic phase (aerobic granular sludge, AGS) and after advanced oxidation processes (AOP) namely ozonation and UV irradiation as post-treatment, were tested to evaluate the fitness of this treatments for ecotoxicity abatement. AGS treatment achieved an 80% removal in both COD and color. AOP was applied with the intention to mineralize the metabolites resulting from biodecolorization of the azo dye Acid Red 14, especially the stable aromatic amine (4-amino-1-naphthalenesulfonic acid, 4A1NS). The ecotoxicity evaluation was based on growth inhibition of the algae Pseudokirchneriella subcapitata following OECD TG 201 except regarding the medium, MBL medium was used instead. Five replicate control cultures and samples were performed with an average STD of 2.7% regarding specific algae growth rate determination. It was found that untreated textile effluent holds an inhibition of specific growth rate of 82%. AGS treatment by itself is able to lower ecotoxicity to 53%. This is probably due to the high color removal of the treatment. AOP post-treatment with Ozone and UV irradiation improves the ecotoxicity abatment to 49 and 43% inhibition respectively, less significantly than previously thought. Since over 85% of 4A1NS was removed by either of the AOP (followed by HPLC), an individual ecotoxicity test of 4A1NS was performed showing that 4A1NS does not inhibit algae growth (0% inhibition). It was concluded that AGS treatment is able by itself to achieve a significant ecotoxicity abatement of textile effluent. The cost-benefit of AOP as a post-treatment have to be better accessed since their application resulted in an improvement of only 10% regarding ecotoxicity effluent removal. It was also found that the 4A1NS amine had no apparent effect on ecotoxicity. Further studies will be conducted to study where ecotoxicity is coming from after AGS biological treatment and how to eliminate it.

Keywords: textile wastewate, ecotoxicity, aerobic granular sludge, AOP

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Authors: Gadanya, A.M., Ponfa, S., Jibril, M.M., Abubakar, S. M.

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Background: Edible insects such as grasshopper are important sources of food for humans, and have been consumed as traditional foods by many indigenous communities especially in Africa, Asia, and Latin America. These communities have developed their skills and techniques in harvesting, preparing, consuming, and preserving edible insects, widely contributing to the role played by the use of insects in human nutrition. Aim/ objective: This study was aimed at determining the effect of insect enriched sprouted soyabean /millet based food on some gut health markers in albino rats. Methods. Four different formulations of Complementary foods (i.e Complementary Food B (CFB): sprouted millet (SM), Complementary Food C (CFC): sprouted soyabean (SSB), Complementary Food D (CFD): sprouted soybean and millet (SSBM) in a ratio of (50:50) and Complementary Food E (CFE): insect (grasshopper) enriched sprouted soybean and millet (SSBMI) in a ratio of (50:25:25)) were prepared. Proximate composition and short chain fatty acid contents were determined. Thirty albino rats were divided into5 groups of six rats each. Group 1(CDA) were fed with basal diet and served as a control group, while groups 2,3,4 and 5 were fed with the corresponding complimentary foods CFB, CFC, CFD and CFE respectively daily for four weeks. Concentrations of fecal protein, serum total carotenoids and nitric oxide were determined. DNA extraction for molecular isolation and characterization were carried out followed by PCR, the use of mega 11 software and NCBI blast for construction of the phylogenetic tree and organism identification respectively. Results: Significant increase (P<0.05) in percentage ash, fat, protein and moisture contents, as well as short chain fatty acid (acetate, butyrate and propionate) concentrations were recorded in the insect enriched sprouted composite food (CFE) when compared with the CFA, CFB, CFC and CFD composite food. Faecal protein, carotenoid and nitric oxide concentrations were significantly lower (P>0.05) in group 5 in comparison to groups 1to 4. Ruminococcus bromii and Bacteroidetes were molecularly isolated and characterized by 16s rRNA from the sprouted millet/sprouted soybean and the insect enriched sprouted soybean/sprouted millet based food respectively. The presence of these bacterial strains in the feaces of the treated rats is an indication that the gut of the treated rats is colonized by good gut bacteria, hence, an improved gut health. Conclusion: Insect enriched sprouted soya bean/sprouted millet based complementary diet showed a high composition of ash, fat, protein and fiber. Thus, could increase the availability of short chain fatty acids whose role to the host organism cannot be overemphasized. It was also found to have decrease the level of faecal protein, carotenoid and nitric oxide in the serum which is an indication of an improvement in the immune system function.

Keywords: gut-health, insect, millet, soybean, sprouted

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Authors: K. Nazan Turhan, Tuğçe Ersan

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The population of the world is approximately 8 billion, and it increases uncontrollably and irrepressibly, leading to an increase in consumption. This situation causes crucial problems, and food waste is one of these. The Food and Agriculture Organization of the United Nations (FAO) defines food waste as the discarding or alternative utilization of food that is safe and nutritious for the consumption of humans along the entire food supply chain, from primary production to end household consumer level. In addition, according to the estimation of FAO, one-third of all food produced for human consumption is lost or wasted worldwide every year. Wasting food endangers natural resources and causes hunger. For instance, excessive amounts of food waste cause greenhouse gas emissions, contributing to global warming. Therefore, waste management has been gaining significance in the last few decades at both local and global levels due to the expected scarcity of resources for the increasing population of the world. There are several ways to recover food waste. According to the United States Environmental Protection Agency’s Food Recovery Hierarchy, food waste recovery ways are source reduction, feeding hungry people, feeding animals, industrial uses, composting, and landfill/incineration from the most preferred to the least preferred, respectively. Bioethanol, biodiesel, biogas, agricultural fertilizer and animal feed can be obtained from food waste that is generated by different food industries. In this project, feeding animals was selected as a food waste recovery method and food waste of a plant was used to provide ingredient uniformity. Grasshoppers were used as a protein source. In other words, the project was performed to develop a dog food product by recovery of the plant’s food waste after following some steps. The collected food waste and purchased grasshoppers were sterilized, dried and pulverized. Then, they were all mixed with 60 g agar-agar solution (4%w/v). 3 different aromas were added, separately to the samples to enhance flavour quality. Since there are differences in the required amounts of different species of dogs, fulfilling all nutritional needs is one of the problems. In other words, there is a wide range of nutritional needs in terms of carbohydrates, protein, fat, sodium, calcium, and so on. Furthermore, the requirements differ depending on age, gender, weight, height, and species. Therefore, the product that was developed contains average amounts of each substance so as not to cause any deficiency or surplus. On the other hand, it contains more protein than similar products in the market. The product was evaluated in terms of contamination and nutritional content. For contamination risk, detection of E. coli and Salmonella experiments were performed, and the results were negative. For the nutritional value test, protein content analysis was done. The protein contents of different samples vary between 33.68% and 26.07%. In addition, water activity analysis was performed, and the water activity (aw) values of different samples ranged between 0.2456 and 0.4145.

Keywords: food waste, dog food, animal nutrition, food waste recovery

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Authors: Loredana E. Nita, Aurica P. Chiriac, Elena Stoleru, Alina Diaconu, Tudorachi Nita

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Self-assembly processes are an attractive method to form new and complex structures between macromolecular compounds to be used for specific applications. In this context, intramolecular and intermolecular bonds play a key role during self-assembling processes in preparation of carrier systems of bioactive substances. Polyelectrolyte complexes (PECs) are formed through electrostatic interactions, and though they are significantly below of the covalent linkages in their strength, these complexes are sufficiently stable owing to the association processes. The relative ease way of PECs formation makes from them a versatile tool for preparation of various materials, with properties that can be tuned by adjusting several parameters, such as the chemical composition and structure of polyelectrolytes, pH and ionic strength of solutions, temperature and post-treatment procedures. For example, protein-polyelectrolyte complexes (PPCs) are playing an important role in various chemical and biological processes, such as protein separation, enzyme stabilization and polymer drug delivery systems. The present investigation is focused on evaluation of the PPC formation between a synthetic polypeptide (poly(aspartic acid) – PAS) and a natural protein (bovine serum albumin - BSA). The PPC obtained from PAS and BSA in different ratio was investigated by corroboration of various techniques of characterization as: spectroscopy, microscopy, thermo-gravimetric analysis, DLS and zeta potential determination, measurements which were performed in static and/or dynamic conditions. The static contact angle of the sample films was also determined in order to evaluate the changes brought upon surface free energy of the prepared PPCs in interdependence with the complexes composition. The evolution of hydrodynamic diameter and zeta potential of the PPC, recorded in situ, confirm changes of both co-partners conformation, a 1/1 ratio between protein and polyelectrolyte being benefit for the preparation of a stable PPC. Also, the study evidenced the dependence of PPC formation on the temperature of preparation. Thus, at low temperatures the PPC is formed with compact structure, small dimension and hydrodynamic diameter, close to those of BSA. The behavior at thermal treatment of the prepared PPCs is in agreement with the composition of the complexes. From the contact angle determination results the increase of the PPC films cohesion, which is higher than that of BSA films. Also, a higher hydrophobicity corresponds to the new PPC films denoting a good adhesion of the red blood cells onto the surface of PSA/BSA interpenetrated systems. The SEM investigation evidenced as well the specific internal structure of PPC concretized in phases with different size and shape in interdependence with the interpolymer mixture composition.

Keywords: polyelectrolyte – protein complex, bovine serum albumin, poly(aspartic acid), self-assembly

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Authors: Karen G. Bastidas Gomez, Hugo R. Zea Ramirez, Manuel F. Ribeiro Pereira, Cesar A. Sierra Avila, Juan A. Clavijo Morales

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The contamination of water sources with heavy metals such as mercury has been an environmental problem; it has generated a high impact on the environment and human health. In countries such as Colombia, mercury contamination due to mining has reached levels much higher than the world average. This work proposes the use of fique fibers as adsorbent in mercury removal. The evaluation of the material was carried out under five different conditions (raw, pretreated by organosolv, functionalized by TEMPO oxidation, fiber functionalized plus MOF-199 and fiber functionalized plus MOF-199-SH). All the materials were characterized using FTIR, SEM, EDX, XRD, and TGA. Regarding the mercury removal, it was done under room pressure and temperature, also pH = 7 for all materials presentations, followed by Atomic Absorption Spectroscopy. The high cellulose content in fique is the main particularity of this lignocellulosic biomass since the degree of oxidation depends on the number of hydroxyl groups on the surface capable of oxidizing into carboxylic acids, a functional group capable of increasing ion exchange with mercury in solution. It was also expected that the impregnation of the MOF would increase the mercury removal; however, it was found that the functionalized fique achieved a greater percentage of removal, resulting in 81.33% of removal, 44% for the fique with the MOF-199 and 72% for the MOF-199-SH with. The pretreated fiber and raw also showed 74% and 56%, respectively, which indicates that fique does not require considerable modifications in its structure to achieve good performances. Even so, the functionalized fiber increases the percentage of removal considerably compared to the pretreated fique, which suggests that the functionalization process is a feasible procedure to apply with the purpose of improving the removal percentage. In addition, this is a procedure that follows a green approach since the reagents involved have low environmental impact, and the contribution to the remediation of natural resources is high.

Keywords: biomass, nanotechnology, science materials, wastewater treatment

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Authors: Syed Asif Hassan, Tabrej Khan

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Multiple sclerosis (MS) is a chronic demyelinating autoimmune disorder, of the central nervous system (CNS). In the present scenario, the current therapies either do not halt the progression of the disease or have side effects which limit the usage of current Disease Modifying Therapies (DMTs) for a longer period of time. Therefore, keeping the current treatment failure schema, we are focusing on screening novel analogues of the available DMTs that specifically bind and inhibit the Sphingosine1-phosphate receptor1 (S1PR1) thereby hindering the lymphocyte propagation toward CNS. The novel drug-like analogs molecule will decrease the frequency of relapses (recurrence of the symptoms associated with MS) with higher efficacy and lower toxicity to human system. In this study, an integrated approach involving ligand-based virtual screening protocol (Ultrafast Shape Recognition with CREDO Atom Types (USRCAT)) to identify the non-toxic drug like analogs of the approved DMTs were employed. The potency of the drug-like analog molecules to cross the Blood Brain Barrier (BBB) was estimated. Besides, molecular docking and simulation using Auto Dock Vina 1.1.2 and GOLD 3.01 were performed using the X-ray crystal structure of Mtb LprG protein to calculate the affinity and specificity of the analogs with the given LprG protein. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has a higher hypothetical affinity, also has greater negative value. Further, the non-specific ligands were screened out using the structural filter proposed by Baell and Holloway. Based on the USRCAT, Lipinski’s values, toxicity and BBB analysis, the drug-like analogs of fingolimod and BG-12 showed that RTL and CHEMBL1771640, respectively are non-toxic and permeable to BBB. The successful docking and DSX analysis showed that RTL and CHEMBL1771640 could bind to the binding pocket of S1PR1 receptor protein of human with greater affinity than as compared to their parent compound (Fingolimod). In this study, we also found that all the drug-like analogs of the standard MS drugs passed the Bell and Holloway filter.

Keywords: antagonist, binding affinity, chemotherapeutics, drug-like, multiple sclerosis, S1PR1 receptor protein

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Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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This study was conducted to determine that carrot powder obtain by different drying methods (oven and vacuum-freeze dryer) of carrot unfit for human consumption that whether feed additives in animal nutrition or not. Carrots randomly divided 2 groups. First group was dried by using oven, second group was by using vacuum freeze dryer methods. Dried carrot prepared from fresh carrot was analysed nutrient matter (energy, crude protein, crude oil, crude ash, beta carotene, mineral concentration and colour). The differences between groups in terms of energy, crude protein, ash, Ca and Mg was not significant (P> 0,05). Crude oil, P, beta carotene content and colour values (L, a, b) with vacuum-freeze dryer group was greater than oven group (P<0,05). Consequently, carrot powder obtained by drying the vacuum-freeze dryer method can be used as a source of carotene.

Keywords: carrot, vacuum freeze dryer, oven, beta carotene

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Authors: Fatima Arrutia, Francisco Amador Riera

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The meat industry generates large volumes of blood as a result of meat processing. Several industrial procedures have been implemented in order to treat this by-product, but are focused on the production of low-value products, and in many cases, blood is simply discarded as waste. Besides, in addition to economic interests, there is an environmental concern due to bloodborne pathogens and other chemical contaminants found in blood. Consequently, there is a dire need to find extensive uses for blood that can be both applicable to industrial scale and able to yield high value-added products. Blood has been recognized as an important source of protein. The main blood serum protein in mammals is serum albumin. One of the top trends in food market is functional foods. Among them, bioactive peptides can be obtained from protein sources by microbiological fermentation or enzymatic and chemical hydrolysis. Bioactive peptides are short amino acid sequences that can have a positive impact on health when administered. The main drawback for bioactive peptide production is the high cost of the isolation, purification and characterization techniques (such as chromatography and mass spectrometry) that make unaffordable the scale-up. On the other hand, membrane technologies are very suitable to apply to the industry because they offer a very easy scale-up and are low-cost technologies, compared to other traditional separation methods. In this work, the possibility of obtaining bioactive peptide fractions from serum albumin by means of a simple procedure of only 2 steps (hydrolysis and membrane filtration) was evaluated, as an exploratory study for possible industrial application. The methodology used in this work was, firstly, a tryptic hydrolysis of serum albumin in order to release the peptides from the protein. The protein was previously subjected to a thermal treatment in order to enhance the enzyme cleavage and thus the peptide yield. Then, the obtained hydrolysate was filtered through a nanofiltration/ultrafiltration flat rig at three different pH values with two different membrane materials, so as to compare membrane performance. The corresponding permeates were analyzed by liquid chromatography-tandem mass spectrometry technology in order to obtain the peptide sequences present in each permeate. Finally, different concentrations of every permeate were evaluated for their in vitro antihypertensive and antioxidant activities though ACE-inhibition and DPPH radical scavenging tests. The hydrolysis process with the previous thermal treatment allowed achieving a degree of hydrolysis of the 49.66% of the maximum possible. It was found that peptides were best transmitted to the permeate stream at pH values that corresponded to their isoelectric points. Best selectivity between peptide groups was achieved at basic pH values. Differences in peptide content were found between membranes and also between pH values for the same membrane. The antioxidant activity of all permeates was high compared with the control only for the highest dose. However, antihypertensive activity was best for intermediate concentrations, rather than higher or lower doses. Therefore, although differences between them, all permeates were promising regarding antihypertensive and antioxidant properties.

Keywords: bioactive peptides, bovine serum albumin, hydrolysis, membrane filtration

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Authors: Arámbula-Villa Gerónimo, García-Lara Kenia Y., Figueroa-Cárdenas J. D., Pérez-Robles J. F., Jiménez-Sandoval S., Salazar-López R., Herrera-Corredor J. A.

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The nixtamalization process (thermal-alkaline method) improves the nutritional part of the corn grain. In this process, the using of Ca(OH)₂ is basic, although the chemical mechanisms between this alkali and the carbohydrates (starch), proteins, lipids, and fiber have not been fully identified. In this study, the native corn starch was taken as a model, and it was subjected to cooking with different concentrations of lime (nixtamalization process) and specific studies of FTIR and XRD were carried out to identify the formation of chemical compounds, and the physical, physicochemical, rheological (paste) and structural properties of material obtained were determined. The FTIR spectra showed the formation of calcium-starch complexes. The treatments with Ca(OH)₂ showed a band shift towards 1675 cm⁻¹ and a band in 1436 cm⁻¹ (COO⁻), indicating the oxidation of starch. Three bands were identified (1575, 1550, and 1540 cm⁻¹) characteristics of carboxylic acid salts for three types of coordinated structures: monodentate, pseudo-bridged, and bidentate. The XRD spectra of starch treated with Ca(OH)₂ showed a peak corresponding to CaCO₃ (29.40°). The oxidation of starch was favored with low concentrations of Ca(OH)₂, producing carboxyl and carbonyl groups and increasing the residual CaCO₃. The increased concentration of Ca(OH)₂ showed the formation of calcium carboxylates, with a decrease in relative crystallinity and residual CaCO₃. Samples with low concentrations of Ca(OH)₂ slowed the onset of gelatinization and increased the swelling of the granules and the peak viscosity. The higher concentrations of Ca(OH)₂ difficulted the water absorption and decreased the viscosity rate and peak viscosity. These results can be used to improve the quality characteristics of the dough and tortillas and to get better acceptance by consumers.

Keywords: maize starch, nixtamalization, gelatinization, calcium carboxylates

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Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

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Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study was performed to examine the effects of different parts of deer antler extract (DH) on osteogenic gene expressions in MG-63 cells and longitudinal bone growth in adolescent male rats. The expressions of osteogenic genes, collagen, alkaline phosphatase, osteocalcin, and osteopontin, were measured by quantitative real-time PCR. Longitudinal bone growth was measured in 3-week-old male Sprague-Dawley rats using fluorescence microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate and bone morphogenetic protein-2 (BMP-2) were measured. Collagen and osteocalcin mRNA expressions were increased by all three parts of the DH treatment while osteopontin gene expression was not affected by any of the DH treatment. Alkaline phosphatase gene expression was increased by upper and mid part of DH while base part of DH fails to affect alkaline phosphatase gene expression. The upper and mid parts of the DH treatment enhanced longitudinal bone growth and total height of growth plate. The induction of BMP-2 protein expression in growth plate assessed by immunostaining was also promoted by upper and mid parts of the DH treatment. These results suggest that DH, especially upper and mid parts, stimulate osteogenic gene expressions and have the effect on bone growth in adolescent rats and might be used for the growth delayed adolescent and inherent growth failure patient.

Keywords: bone morphogenetic protein-2, deer antler, longitudinal bone growth, osteogenic genes

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Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

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Authors: Gelu Varghese, Khizer Saeed

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Biodiesel is as a mixture of mono-alkyl esters of long chain fatty acids derived from vegetable oils or animal fats. Recent studies in the literature suggest that end property of biodiesel such as its oxidative stability (OS) is highly influenced by the structure and composition of its alkyl esters than by environmental conditions. The structure and composition of these long chain fatty acid components have been also associated with trends in Cetane number, heat of combustion, cold flow properties viscosity, and lubricity. In the present work, detailed investigation has been carried out to decouple and correlate the fatty acid structure indices of biodiesel such as degree of unsaturation, chain length, double bond orientation, and composition with its oxidative stability. Measurements were taken using the EN14214 established Rancimat oxidative stability test method (EN141120). Firstly, effects of the degree of unsaturation, chain length and bond orientation were tested for the pure fatty acids to establish their oxidative stability. Results for pure Fatty acid show that Saturated FAs are more stable than unsaturated ones to oxidation; superior oxidative stability can be achieved by blending biodiesel fuels with relatively high in saturated fatty acid contents. A lower oxidative stability is noticed when a greater quantity of double bonds is present in the methyl ester. A strong inverse relationship with the number of double bonds and the Rancimat IP values can be identified. Trans isomer Methyl elaidate shows superior stability to oxidation than its cis isomer methyl oleate (7.2 vs. 2.3). Secondly, the effects of the variation in the composition of the biodiesel were investigated and established. Finally, biodiesels with varying structure and composition were investigated and correlated.

Keywords: biodiesel, fame, oxidative stability, fatty acid structure, acid composition

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Authors: Masataka Inada, Masanao Kinoshita, Nobuaki Matsumori

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It has been demonstrated that lipid molecules in biological membranes are responsible for the functionalization and structuration of membrane proteins. However, it is still unclear how the interaction of lipid molecules with membrane proteins is correlated with the function of the membrane proteins. Here we first developed an evaluation method for the interaction between membrane proteins and lipid molecules via surface plasmon resonance (SPR) analysis. Bacteriorhodopsin (bR), which was obtained by the culture of halobacteria, was used as a membrane protein. We prepared SPR sensor chips covered with self-assembled monolayer containing mercaptocarboxylic acids, and immobilized bR onto them. Then, we evaluated the interactions with various lipids that have different structures. As a result, the halobacterium-specific glycolipid S-TGA-1 was found to have much higher affinity with bRs than other lipids. This is probably due to not only hydrophobic and electrostatic interactions but also hydrogen bonds with sugar moieties in the glycolipid. Next, we analyzed the roles of the lipid in the structuration and functionalization of bR. CD analysis showed that S-TGA-1 could promote trimerization of bR monomers more efficiently than any other lipids. Flash photolysis further indicated that bR trimers formed by S-TGA-1 reproduced the photocyclic activity of bR in purple membrane, halobacterium-membrane. These results suggest that S-TGA-1 promotes trimerization of bR through strong interactions and consequently fulfills the bR’s function efficiently.

Keywords: membrane protein, lipid, interaction, bacteriorhodopsin, glycolipid

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Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang

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Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).

Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells

Procedia PDF Downloads 269