Search results for: fish sarcoplasmic protein

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: 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: Manisha, Manisha Mathur, Jay K. Desai, Shesh Asopa, Manisha Mehra

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The present study was carried out for the comprehensive analysis of lumpy skin disease (LSD) in cattle and to elucidate the histopathology of vital organs in natural outbreaks. Lumpy skin disease (LSD) is a viral infection that primarily affects cattle. It is caused by a Capri pox virus and is characterized by the formation of skin nodules or lesions. For this study, a postmortem of 20 cows who died of Lumpy skin disease in different regions of Rajasthan was conducted. This study aimed to examine a cow's external and internal organs to confirm if lumpy skin disease was the cause of death. Accurate diagnosis is essential for improving disease surveillance, understanding the disease's progression, and informing control measures. Pathological examinations reveal virus-induced changes across organs, while histopathological analyses provide crucial insights into the disease's pathogenesis, aiding in the development of advanced diagnostics and effective prevention strategies. Histopathological examination of nodular skin lesions revealed edema, hyperemia, acanthosis, severe hydropic degeneration/ballooning degeneration, and hyperkeratosis in the epidermis. In the lungs, congestion, oedema, emphysema, and atelectasis were observed grossly. Microscopically changes were suggestive of interstitial pneumonia, suppurative pneumonia, bronchopneumonia post pneumonic fibrosis, and stage of resolution. Grossely liver showed congestion and necrotic foci microscopically in most of the cases, and the liver showed acute viral hepatitis. Microscopically in kidneys, multifocal interstitial nephritis was observed. There was marked interstitial inflammation and zonal fibrosis with cystically dilated tubules and bowman's capsules. Microscopically, most of the heart tissue section showed normal histology with few sarcocysts in between cardiac muscles. In some cases, loss of cross striation, sarcoplasmic vacuolation, fregmentation, and disintegration of cardiac fibres were observed. The present study revealed the characteristic gross and histopathological changes in different organs in natural cases of lumpy skin disease. Further, the disease was confirmed based on the molecular diagnosis and transmission electron microscopy of capripox infection in the affected cattle in the study area.

Keywords: Capripoxvirus, lumpy skin disease, polymerage chain reaction, transmission electron microscopy

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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: Edwin O. Clarke, Akintade O. Adeboyejo

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Intensive use of pesticides resulted in dispersal of pollutants throughout the globe. This study was carried out to investigate persistent Organochlorine pesticides (POPs) in water, sediment and fin fishes, Schilbes mystus and Hemichromis fasciatus from two different sampling stations along River Ogun between the month of June 2012 and January 2013. The Organochlorine pesticides analyzed include DDT (pp’1,1,1-trichloro-2,2-bis-(4-chlorophenyl) ethane), DDD, DDE (pp1,1-dichloro-2, 2-bis-(4-chlorophenyl) ethylene, HCH (gamma 1,2,3,4,5,6-hexachlorocylohexane, HCB hexachlorobenzene),Dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a octahydro- 1,4,5,8 dimethanonaphthalene). The analysis was done using Gas Chromatograph with Electron Capture Detector. In water sample, the result showed that PPDDT, Endrin aldehyde, Endrin ketone concentrations were high in both stations. The mean value of Organochlorine analyzed in water range from Beta BHC (0.50±0.10µg/l) to PP DDT (162.86±0.21µg/l) in Kara sample station and Beta BHC (0.20±0.07µg/l) to Endrin Aldehyde (76.47±0.02µg/l) in Odo-Ogun sample station. The levels of POPs obtained in sediments ranged from 0.40±0.23µg/g (Beta BHC) to 259.90 ± 1.00µg/kg (Endosulfan sulfate) in Kara sample station and 0.64±0.00µg/g (Beta BHC) to 379.77 ±0.15 µg/g (Endosulfan sulfate) in Odo-Ogun sample station. The levels of POPs obtained in fin fish samples ranged from 0.29±0.00µg/g (Delta BHC) to 197.87 ± 0.31µg/g (PP DDT) in Kara sample station and in Odo-Ogun sample station the mean value for fish samples range from 0.29 ± 0.00 µg/g (Delta BHC) to 197.87 ± 0.32 µg/g (PP DDT). The study showed that the accumulation of POPs affect the environment and reduce water quality. The results showed that the concentrations were found to exceed the maximum acceptable concentration of 0.10µg/l value set by the European Union for the protection of freshwater aquatic life and this can be hazardous if the trend is not checked.

Keywords: hazardous, persistent, pesticides, biomes

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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: Sahana Selladurai, Christine DeWolf

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Lung surfactant comprises a lipid-protein film that coats the alveolar surface and serves to prevent alveolar collapse upon repeated breathing cycles. Exposure of lung surfactant to high concentrations of airborne pollutants, for example tropospheric ozone in smog, can chemically modify the lipid and protein components. These chemical changes can impact the film functionality by decreasing the film’s collapse pressure (minimum surface tension attainable), altering it is mechanical and flow properties and modifying lipid reservoir formation essential for re-spreading of the film during the inhalation process. In this study, we use Langmuir monolayers spread at the air-water interface as model membranes where the compression and expansion of the film mimics the breathing cycle. The impact of ozone exposure on model lung surfactant films is measured using a Langmuir film balance, Brewster angle microscopy and a pendant drop tensiometer as a function of film and sub-phase composition. The oxidized films are analyzed using mass spectrometry where lipid and protein oxidation products are observed. Oxidation is shown to reduce surface activity, alter line tension (and film morphology) and in some cases visibly reduce the viscoelastic properties of the film when compared to controls. These reductions in functionality of the films are highly dependent on film and sub-phase composition, where for example, the effect of oxidation is more pronounced when using a physiologically relevant buffer as opposed to water as the sub-phase. These findings can lead to a better understanding on the impact of continuous exposure to high levels of ozone on the mechanical process of breathing, as well as understanding the roles of certain lung surfactant components in this process.

Keywords: lung surfactant, oxidation, ozone, viscoelasticity

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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

Abstract:

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: Lauren A. Bailey, Amber R. Childs, Nicola C. James, Murray I. Duncan, Alexander Winkler, Warren M. Potts

Abstract:

For the persistence of fished populations in the Anthropocene, it is critical to predict how fished populations will respond to the coupled threats of exploitation and climate change for adaptive management. The resilience of fished populations will depend on their capacity for physiological plasticity and acclimatization in response to environmental shifts. However, there is evidence for the selection of physiological traits by capture fisheries. Hence, fish populations may have a limited scope for the rapid expansion of their tolerance ranges or physiological adaptation under fishing pressures. To determine the physiological vulnerability of fished populations in the Anthropocene, the metabolic performance was compared between a fished and spatially protected Chrysoblephus laticeps population in response to thermal variability. Individual aerobic scope phenotypes were quantified using intermittent flow respirometry by comparing changes in energy expenditure of each individual at ecologically relevant temperatures, mimicking variability experienced as a result of upwelling and downwelling events. The proportion of high and low-performance individuals were compared between the fished and spatially protected population. The fished population had limited aerobic scope phenotype diversity and fewer high-performance phenotypes, resulting in a significantly lower aerobic scope curve across low (10 °C) and high (24 °C) thermal treatments. The performance of fished populations may be compromised with predicted future increases in cold upwelling events. This requires the conservation of the physiologically fittest individuals in spatially protected areas, which can recruit into nearby fished areas, as a climate resilience tool.

Keywords: climate change, fish physiology, metabolic shifts, over-fishing, respirometry

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Authors: Syed Asif Hassan, Tabrej Khan

Abstract:

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

Abstract:

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

Abstract:

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: Roger G. Pagente Jr.

Abstract:

This study seeks to investigate the language learning experiences of the thirty-nine AB-English majors who were selected through fish-bowl technique from the 157 students enrolled in the AB-English program. Findings taken from the diary, questionnaire and unstructured interview revealed that motivation, learners’ belief, self-monitoring, language anxiety, activities and strategies were the prevailing factors that influenced the learning of English of the participants.

Keywords: diary, English language learning experiences, self-monitoring, language anxiety

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

Abstract:

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

Abstract:

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: Masataka Inada, Masanao Kinoshita, Nobuaki Matsumori

Abstract:

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

Procedia PDF Downloads 253

Authors: M. B. Mshelia, J. K Balogun, J. Auta, N. O. Bankole

Abstract:

The current status and abundance of the genus Citharinus was carried out in Jebba Lake, Niger State, Nigeria from January to December, 2011. The aim was to determine the extent of exploitation of the genus Citharinus in Jebba Lake so as to advice the government of Nigeria on how to overcome difficulties in terms of the sustainability of the said fish in the Lake. Descriptive statistics were used to analyze the data obtained. A total of 2,389 of the genus Citharinus were caught during the sampling period. Only two species of the genus Citharinus were caught with 1,220 in number and 430.68kg total weight of Citharinus citharus and 1,169 in number and 418.56kg total weight of Citharinus latus). The current total yield estimated for the genus Citharinus in Jebba Lake in the six (6) sampling sites was calculated and pooled together to be 849.24kg. A day’s catch was calculated to be 35.38kg. The monthly and annual yields of the genus Citharinus were calculated to 1061.55 equivalents to 1 ton and 12 metric tonnes respectively. For the fecundity, June, July and August were discovered as the spawning period for the genus Citharinus and out of total experimental gillnet catch of 2, 389, only 244 (10.21%)of Citharinus citharus and 231 (9.67%) of Citharinus latus were in sexually mature stage. Out of these numbers, 113 (46.31%) were males and 121 (53.69%) were females of Citharinus citharus and 112 (48.48) were males and 119 (51.52) were females of Citharinus latus. The youngest mature males in either of the two had a standard length of 31.5 with a weight of 800.5gWhilethe youngest spawning females were having the standard length of 29.5 cm with a weight of 1,3oo.5g.It was also discovered that females started maturing earlier than the males at the standard length for females and males to be 18.0cm and 19.5cm respectively. Their fecundity ranged from 15,000 to 16, 500 eggs. The sex ratio of 1172 that were males and 1217 that were females was 1 male to 1.0383 females which was equivalent to 1:1 sex ratio of male to female. It was concluded that Jebba Lake had suffered seriously over exploitation of the genus Citharinus and proper management have to be enforced on the lake otherwise the threat of fish being extent may arise.

Keywords: Jebba Lake, Niger State, Nigeria, Citharinus citharus, Citharinus latus, fecundity, sex ratio

Procedia PDF Downloads 261

Authors: Hossein Khara, Mahmoud Sayyadborani, Mohammad Sayyadborani

Abstract:

In the present study, we examined the effects of different dietary levels of ascorbic acid (vitamin C) and α-tocopherol (vitamin E) and their combinations on growth, survival and some haematological and immunological parameters of Caspian brown trout, Salmo trutta caspius juveniles. 15 experimental treatments and one control group with three replicates were considered for experiment. The experimental treatments were fish fed by experimental diets containing different levels of Vit C and E as follow: T1: Vit E (20 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T2: Vit E (30 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T3: Vit E (40 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T4: Vit E (20 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T5: Vit E (30 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T6: Vit E (40 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T7: Vit E (20 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T8: Vit E (30 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T9: Vit E (40 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T10: Vit C (100 mg.kg diet -1), T11: Vit C (200 mg.kg diet -1), T12: Vit C (300 mg.kg diet -1), T13: Vit E (20 mg.kg diet -1), T14: Vit E (30 mg.kg diet -1) T15: Vit E (40 mg.kg diet -1). Also a non-vitamin supplemented was considered as control group. Growth parameters were measured monthly and serum parameters assayed at the end of the experiment. According to our results, Vit C and E improved survival and growth parameters including specific growth rate (SGR), weight gain percent (WG%) and biomass. The highest values of these parameters obtained in T8, T9 and T8 respectively. The lowest FCR obtained in T8. The haematological parameters including red blood cells (RBCs), white blood cells (WBCs), haematocrit (Hct) and haemoglobin (Hb) were higher in vitamin treated groups than control group with highest values in T8. In T13, WBC values were higher compared to other experimental groups. The immunological parameters including lysozyme activity, Immunoglobulin (IgM) and total immunoglobulin (TIg) were significantly higher in vitamin supplemented groups than in control group. In this regard the highest values of these parameters were found in T12. The lowest values of TIg and lysozyme activity were observed in control group and fish fed by only vitamin E i.e. T13, T14 and T15. In conclusion, our results show that Vit C and E in combination or only can improve growth, survival, haematological and immunological indices of Caspian brown trout.

Keywords: vitamins E, vitamins C, growth, survival, haematological parameters, immunological parameters

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Authors: Sali Chinsathit, Haruthai Kaenla

Abstract:

Organic agriculture has become a major policy theme for agricultural development in Thailand since October 2005. Organic farming is enlisted as an important national agenda, to promote safe food and national export, and many government authorities have initiated projects and activities centered on organic farming promotion. Currently, Thailand has the market share of about 32 million US$ a year by exporting organic products of rice, vegetables, tea, fruits and a few medicinal herbs. There is high potential in organic crop production as there is the tropical environment promoting crop growth and leader farmer in organic farming. However, organic sector is relatively small (0.2%) comparing with conventional agricultural area, since there are many factors affecting farmers’ adoption and success in organic farming. The objective of this project was to get the organic production technology for at least 3 organic crops. The treatment and method were complied with Thai Organic Standard, and were mainly concerned on increase plant biodiversity and soil improvement by using organic fertilizer and bio-extract from fish, egg, plant and fruits. The bio-logical control, plant-extracts, and cultural practices were used to control insect pests and diseases of 3 crops including mangosteen (Garcinia mangostana L.), longkong (Aglaia dookoo Griff.) and banana (Musa (AA group)). The experiments were carried out at research centers of Department of Agriculture and farmers’ farms in Rayong and Chanthaburi provinces from 2009 to 2013. We found that both locations, plant biodiversity by intercropping mangosteen or longkong with banana and soil improvement with composts and bio-extract from fish could increased yield and farmers’ income by 6,835 US$/ha/year. Farmers got knowledge from these technologies to produce organic crops. The organic products were sold both in domestic and international countries. The organic production technologies were also environmental friendly and could be used as an alternative way for farmers in Thailand.

Keywords: banana, longkong, mangosteen, organic farming

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

Abstract:

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

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Authors: Akshita Bhardwaj

Abstract:

Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.

Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density

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Authors: Aqeel Ahmed Shad, Tanveer Ahmad, Muhammad Farooq Iqbal, Muhammad Javaid Asad

Abstract:

The supplemental effects of novel protease produced from Bacillus subtilis K-5 and beta-mannanase were evaluated on growth performance, carcass characteristics, nutrients digestibility, blood profile and intestinal morphometry of broilers fed guar meal (Cyamopsis tetragonoloba) based diets with reduced Crude Protein (CP), Essential Amino Acids (EAAs), and Metabolizable energy (ME) contents. One-day old Ross 308 broiler chicks (n=360) were randomly allotted to thirty six experimental units in a way that each of the nine dietary treatments received four replicates with ten birds per replicate. A control diet without guar meal (0GM) was formulated with standard nutrient specifications of Ross 308 for the starter and finisher phases. Two negative control diets, one with 5% (5GM) and second with 10% (10GM) guar meal, were formulated with reduction of 5% CP, 5% EAAs and 80 Kcal/kg ME. These three basal diets (no enzyme) were supplemented with novel protease enzyme (PROT) and commercial beta-mannanase (Beta-M) enzyme. The birds were reared up to 35d of age. The data on weekly body weight gain (BWG) and feed intake were recorded to compute feed:gain for the starter (0-21d) and finisher (22-35d) phases. At the end of 35d of experimental period, four birds per experimental unit were randomly selected for blood samples collection and later slaughtered for ileal digesta, intestinal tract and carcass trait sampling. The data on overall performance (1-35d) indicated improved (P<0.05) BWG and feed:gain in birds supplemented with PROT (1.41% and 1.67) and Beta-M (2.79% and 1.64) than non-supplemented groups. Improved (P<0.05) carcass yield, breast meat yield and thigh meat yield were noted with the supplementation of Beta-M. However, non-significant (P>0.05) effect on carcass traits was noted in broiler fed guar meal based PROT supplemented diets. Crude protein digestibility, nitrogen retention (Nret) and apparent digestibility coefficient for nitrogen (ADCN) were improved (P<0.05) only with PROT. The improvement in apparent metabolizable energy (AME) and apparent metabolizable energy corrected for nitrogen (AMEn) was noted (P<0.05) with both supplemented enzymes. However, no effect (P>0.05) of enzyme addition was noted on blood glucose, total protein and cholesterol. Improved villus height of duodenum, jejunum and ileum was noted (P<0.05) with the addition of both enzymes. The EAAs digestibility was improved (P<0.05) only with PROT. In conclusion, beta-mannanase and protease supplementation better improved the overall bird performance in low nutrient profile guar meal based diets than non-supplemented diets.

Keywords: novel protease, guar meal, broilers, low protein diets, low metabolizable energy diets, nutrients digestibility

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