Search results for: acyl-coa binding protein (ACBP)

Authors: Fei Jia, Xingjian Bai, Xiaowei Zhang, Wenjie Yan, Ruitong Dai, Xingmin Li, Jozef Kokini

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Pseudomonas aeruginosa is a Gram-negative, aerobic, opportunistic human pathogen that is present in the soil, water, and food. This microbe has been recognized as a representative food-borne spoilage bacterium that can lead to many types of infections. Considering the casualties and property loss caused by P. aeruginosa, the development of a rapid and reliable technique for the detection of P. aeruginosa is crucial. The whole-cell aptasensor, an emerging biosensor using aptamer as a capture probe to bind to the whole cell, for food-borne pathogens detection has attracted much attention due to its convenience and high sensitivity. Here, a low-field magnetic resonance imaging (LF-MRI) aptasensor for the rapid detection of P. aeruginosa was developed. The basic detection principle of the magnetic relaxation switch (MRSw) nanosensor lies on the ‘T₂-shortening’ effect of magnetic nanoparticles in NMR measurements. Briefly speaking, the transverse relaxation time (T₂) of neighboring water protons get shortened when magnetic nanoparticles are clustered due to the cross-linking upon the recognition and binding of biological targets, or simply when the concentration of the magnetic nanoparticles increased. Such shortening is related to both the state change (aggregation or dissociation) and the concentration change of magnetic nanoparticles and can be detected using NMR relaxometry or MRI scanners. In this work, two different sizes of magnetic nanoparticles, which are 10 nm (MN₁₀) and 400 nm (MN₄₀₀) in diameter, were first immobilized with anti- P. aeruginosa aptamer through 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) chemistry separately, to capture and enrich the P. aeruginosa cells. When incubating with the target, a ‘sandwich’ (MN₁₀-bacteria-MN₄₀₀) complex are formed driven by the bonding of MN400 with P. aeruginosa through aptamer recognition, as well as the conjugate aggregation of MN₁₀ on the surface of P. aeruginosa. Due to the different magnetic performance of the MN₁₀ and MN₄₀₀ in the magnetic field caused by their different saturation magnetization, the MN₁₀-bacteria-MN₄₀₀ complex, as well as the unreacted MN₄₀₀ in the solution, can be quickly removed by magnetic separation, and as a result, only unreacted MN₁₀ remain in the solution. The remaining MN₁₀, which are superparamagnetic and stable in low field magnetic field, work as a signal readout for T₂ measurement. Under the optimum condition, the LF-MRI platform provides both image analysis and quantitative detection of P. aeruginosa, with the detection limit as low as 100 cfu/mL. The feasibility and specificity of the aptasensor are demonstrated in detecting real food samples and validated by using plate counting methods. Only two steps and less than 2 hours needed for the detection procedure, this robust aptasensor can detect P. aeruginosa with a wide linear range from 3.1 ×10² cfu/mL to 3.1 ×10⁷ cfu/mL, which is superior to conventional plate counting method and other molecular biology testing assay. Moreover, the aptasensor has a potential to detect other bacteria or toxins by changing suitable aptamers. Considering the excellent accuracy, feasibility, and practicality, the whole-cell aptasensor provides a promising platform for a quick, direct and accurate determination of food-borne pathogens at cell-level.

Keywords: magnetic resonance imaging, meat spoilage, P. aeruginosa, transverse relaxation time

Procedia PDF Downloads 153

Authors: A. Aslihan Gokaltun, Martin L. Yarmush, Ayse Asatekin, O. Berk Usta

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Poly(dimethylsiloxane) (PDMS) is one of the most widely used materials in the fabrication of microfluidic devices. It is easily patterned and can replicate features down to nanometers. Its flexibility, gas permeability that allows oxygenation, and low cost also drive its wide adoption. However, a major drawback of PDMS is its hydrophobicity and fast hydrophobic recovery after surface hydrophilization. This results in significant non-specific adsorption of proteins as well as small hydrophobic molecules such as therapeutic drugs limiting the utility of PDMS in biomedical microfluidic circuitry. While silicon, glass, and thermoplastics have been used, they come with problems of their own such as rigidity, high cost, and special tooling needs, which limit their use to a smaller user base. Many strategies to alleviate these common problems with PDMS are lack of general practical applicability, or have limited shelf lives in terms of the modifications they achieve. This restricts large scale implementation and adoption by industrial and research communities. Accordingly, we aim to tailor biocompatible PDMS surfaces by developing a simple and one step bulk modification approach with novel smart materials to reduce non-specific molecular adsorption and to stabilize long-term cell analysis with PDMS substrates. Smart polymers that blended with PDMS during device manufacture, spontaneously segregate to surfaces when in contact with aqueous solutions and create a < 1 nm layer that reduces non-specific adsorption of organic and biomolecules. Our methods are fully compatible with existing PDMS device manufacture protocols without any additional processing steps. We have demonstrated that our modified PDMS microfluidic system is effective at blocking the adsorption of proteins while retaining the viability of primary rat hepatocytes and preserving the biocompatibility, oxygen permeability, and transparency of the material. We expect this work will enable the development of fouling-resistant biomedical materials from microfluidics to hospital surfaces and tubing.

Keywords: cell culture, microfluidics, non-specific protein adsorption, PDMS, smart polymers

Procedia PDF Downloads 294

Authors: O. Squillace, R. L. Thompson

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Migration of small molecules to, and across the surface of polymer matrices is a little-studied problem with important industrial applications. Tackifiers in adhesives, flavors in foods and binding agents in paints all present situations where the function of a product depends on the ability of small molecules to migrate through a polymer matrix to achieve the desired properties such as softness, dispersion of fillers, and to deliver an effect that is felt (or tasted) on a surface. It’s been shown that the chemical and molecular structure, surface free energies, phase behavior, close environment and compatibility of the system, influence the migrants’ motion. When differences in behavior, such as occurrence of segregation to the surface or not, are observed it is then of crucial importance to identify and get a better understanding of the driving forces involved in the process of molecular migration. In this aim, experience is meant to be allied with theory in order to deliver a validated theoretical and computational toolkit to describe and predict these phenomena. The systems that have been chosen for this study aim to address the effect of polarity mismatch between the migrants and the polymer matrix and that of a second migrant over the first one. As a non-polar resin polymer, polyvinyl acetate is used as the material to which more or less polar migrants (sorbitol, carvone, octanoic acid (OA), triacetin) are to be added. Through contact angle measurement a surface excess is seen for sorbitol (polar) mixed with PVAc as the surface energy is lowered compare to the one of pure PVAc. This effect is increased upon the addition of carvon or triacetin (non-polars). Surface micro-structures are also evidenced by atomic force microscopy (AFM). Ion beam analysis (Nuclear Reaction Analysis), supplemented by neutron reflectometry can accurately characterize the self-organization of surfactants, oligomers, aromatic molecules in polymer films in order to relate the macroscopic behavior to the length scales that are amenable to simulation. The nuclear reaction analysis (NRA) data for deuterated OA 20% shows the evidence of a surface excess which is enhanced after annealing. The addition of 10% triacetin, as a second migrant, results in the formation of an underlying layer enriched in triacetin below the surface excess of OA. The results show that molecules in polarity mismatch with the matrix tend to segregate to the surface, and this is favored by the addition of a second migrant of the same polarity than the matrix. As studies have been restricted to materials that are model supported films under static conditions in a first step, it is also wished to address the more challenging conditions of materials under controlled stress or strain. To achieve this, a simple rig and PDMS cell have been designed to stretch the material to a defined strain and to probe these mechanical effects by ion beam analysis and atomic force microscopy. This will make a significant step towards exploring the influence of extensional strain on surface segregation, flavor release in cross-linked rubbers.

Keywords: polymers, surface segregation, thin films, molecular migration

Procedia PDF Downloads 134

Authors: Parminder Nain, Jaspreet kaur, Vipin Saini, Sunil Sharma

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Jasminum humile commonly known as yellow Jasmine or Pili chameli, is a medicinal plant used in Ayurveda for treating various diseases, one of which is diabetes mellitus. The current study aimed to establish the antidiabetic and antioxidant properties of aqueous extract of Jasminum humile leaves (AEJHL) in nicotinamide/streptozotocin induced type 2 diabetic rats. Phytochemical screening, HPLC analysis, and acute toxicity study of AEJHL were carried out. Male albino wistar rats (n=42) were divided into seven equal groups. Rats with moderate diabetes having hyperglycemia (blood glucose 250-400 mg/dl) were taken for the experiment. Various concentrations of aqueous extract of Jasminum humile leaves (50, 100, 200 and 300 mg/kg, p.o.), and glibenclamide (1mg/kg, p.o.) were orally administered to diabetic rats for 45 days. The effect of AEJHL on blood glucose, plasma insulin and biochemical parameters such as hemoglobin, total protein, serum creatinine, serum urea, alkaline phosphate, Glutamic-oxalacetic transaminase (SGOT) and glutamic-pyruvic transaminase (SGPT), as well as total cholesterol, triglycerides, and high-density lipoprotein (HDL) were also studied. The antioxidant effect of AEJHL was determined by analyzing hepatic and renal antioxidant markers, like superoxide dismutase (SOD), catalase (CAT), reduced Glutathione (GSH), Glutathione peroxidase (GPx), and lipid peroxidation (LPO) in diabetic rats. After 45-days oral administration of aqueous extract of Jasminum humile leaves significantly (p<0.05) reduced blood sugar and increase plasma insulin level and also reverse all above biochemical parameters and antioxidant enzyme level at dose dependent manner. These findings provide in vivo evidence that the aqueous extract of Jasminum humile leaves possess significant antidiabetic and antioxidant potential in nicotinamide/streptozotocin-induced type-2 diabetes mellitus in rats.

Keywords: antidiabetic, antioxidant, jasminum humile, nicotinamide/streptozotocin, type-2 diabetic

Procedia PDF Downloads 200

Authors: Mustafa M. Donma, Orkide Donma

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Metabolic syndrome (MetS) is a disease associated with obesity. It is a complicated clinical problem possibly affecting body composition as well as macrominerals. These parameters gain further attention, particularly in the pediatric population. The aim of this study is to investigate the amount of discrete body composition fractions in groups that differ in the severity of obesity. Also, the possible associations with calcium (Ca), phosphorus (P), magnesium (Mg) will be examined. The study population was divided into four groups. Twenty-eight, 29, 34, and 34 children were involved in Group 1 (healthy), 2 (obese), 3 (morbid obese), and 4 (MetS), respectively. Institutional Ethical Committee approved the study protocol. Informed consent forms were obtained from the participants. The classification of obese groups was performed based upon the recommendations of the World Health Organization. Metabolic syndrome components were defined. Serum Ca, P, Mg concentrations were measured. Within the scope of body composition, fat mass, fat-free mass, protein mass, mineral mass were determined by a body composition monitor using bioelectrical impedance analysis technology. Weight, height, waist circumference, hip circumference, head circumference, and neck circumference values were recorded. Body mass index, diagnostic obesity notation model assessment index, fat mass index, and fat-free mass index values were calculated. Data were statistically evaluated and interpreted. There was no statistically significant difference among the groups in terms of Ca and P concentrations. Magnesium concentrations differed between Group 1 and Group 4. Strong negative correlations were detected between P as well as Mg and fat mass index as well as diagnostic obesity notation model assessment index in Group 4, the group, which comprised morbid obese children with MetS. This study emphasized unique associations of P and Mg minerals with diagnostic obesity notation model assessment index and fat mass index during the evaluation of morbid obese children with MetS. It was also concluded that diagnostic obesity notation model assessment index and fat mass index were more proper indices in comparison with body mass index and fat-free mass index for the purpose of defining body composition in children.

Keywords: children, fat mass, fat-free mass, macrominerals, obesity

Procedia PDF Downloads 154

Authors: Aurica P. Chiriac, Vera Balan, Mihai Asandulesa, Elena Butnaru, Nita Tudorachi, Elena Stoleru, Loredana E. Nita, Iordana Neamtu, Alina Diaconu, Liliana Mititelu-Tartau

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The interest for studying ‘smart’ materials is entirely justified and in this context were realized investigations on poly(2-hydroxyethylmethacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane), which is a macromolecular compound with sensibility at pH and temperature, gel formation capacity, binding properties, amphilicity, good oxidative and thermal stability. Physico-chemical characteristics in terms of the molecular weight, temperature-sensitive abilities and thermal stability, as well rheological, dielectric and spectroscopic properties were evaluated in correlation with further coupling capabilities. Differential scanning calorimetry investigation indicated Tg at 36.6 °C and a melting point at Tm=72.8°C, for the studied copolymer, and up to 200oC two exothermic processes (at 99.7°C and 148.8°C) were registered with losing weight of about 4 %, respective 19.27%, which indicate just processes of thermal decomposition (and not phenomena of thermal transition) owing to scission of the functional groups and breakage of the macromolecular chains. At the same time, the rheological studies (rotational tests) confirmed the non-Newtonian shear-thinning fluid behavior of the copolymer solution. The dielectric properties of the copolymer have been evaluated in order to investigate the relaxation processes and two relaxation processes under Tg value were registered and attributed to localized motions of polar groups from side chain macromolecules, or parts of them, without disturbing the main chains. According to literature and confirmed as well by our investigations, β-relaxation is assigned with the rotation of the ester side group and the γ-relaxation corresponds to the rotation of hydroxy- methyl side groups. The fluorescence spectroscopy confirmed the copolymer structure, the spiroacetal moiety getting an axial conformation, more stable, with lower energy, able for specific interactions with molecules from environment, phenomena underlined by different shapes of the emission spectra of the copolymer. Also, the copolymer was used as template for indomethacin incorporation as model drug, and the biocompatible character of the complex was confirmed. The release behavior of the bioactive compound was dependent by the copolymer matrix composition, the increasing of 3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane comonomer amount attenuating the drug release. At the same time, the in vivo studies did not show significant differences of leucocyte formula elements, GOT, GPT and LDH levels, nor immune parameters (OC, PC, and BC) between control mice group and groups treated just with copolymer samples, with or without drug, data attesting the biocompatibility of the polymer samples. The investigation of the physico-chemical characteristics of poly(2-hydrxyethyl methacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane) in terms of temperature-sensitive abilities, rheological and dielectrical properties, are bringing useful information for further specific use of this polymeric compound.

Keywords: bioapplications, dielectric and spectroscopic properties, dual sensitivity at pH and temperature, smart materials

Procedia PDF Downloads 282

Authors: H. Haseena Banu, D. Karthick, R. Stalin, E. Nandha Kumar, T. P. Sachidanandam, P. Shanthi

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Background of the study: Type 2 diabetes is emerging as the predominant metabolic disorder in the world among adults characterized mainly by the resistance of the insulin sensitive tissues towards insulin followed by the decrease in the insulin secretion. The treatment for this disease usually involves treatment with oral synthetic drugs which are known to cause several side effects. Therefore, identification of new biomarkers as therapeutic target is the need of the hour. miRNAs are small, non–protein-coding RNAs that negatively regulate gene expression by promoting degradation and/or inhibit the translation of target mRNAs and have emerged as biomarkers in predicting diabetes mellitus. Objective of the study: To elucidate the therapeutic role of gallic acid in modulating the alterations in glucose metabolism induced by miRNAs 194 and 135a in Type 2 diabetic rats. Materials and Methods: T2D was induced in rats by feeding them with a high fat diet for 2 weeks followed by intraperitoneal injection of 35 mg/kg/body weight (b.wt.) of streptozotocin. Microarrays were used to assess the expression of miRNAs in control, diabetic and gallic acid treated rats. Gene expression studies were carried out by RT PCR analysis. Results: Forty one miRNAs were differentially expressed in Type 2 diabetic rats. Among these, the expression of miRNA 194 was significantly decreased whereas miRNA 135a was significantly increased in Type 2 diabetic rats. The glucose metabolism was also altered significantly in skeletal muscle of Type 2 diabetic rats. Conclusion: T2D is associated with alterations in the expression of miRNAs in skeletal muscle. Both these miRNAs 194 and 135a play an important role in glucose metabolism in skeletal muscle of diabetic rats. Gallic acid effectively ameliorated the alterations in glucose metabolism. Hence, both these miRNAs can serve as biomarkers and therapeutic targets in diabetes mellitus. The study also establishes the role of gallic acid as therapeutic agent. Acknowledgment: The financial assistance provided in the form of ICMR women scientist by ICMR DHR INDIA is gratefully acknowledged here.

Keywords: gallic acid, high fat diet, type 2 diabetes mellitus, miRNAs

Procedia PDF Downloads 350

Authors: Sheraz Gul

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The requirements for progressing a compound to clinical trials is well established and relies on the results from in-vitro and in-vivo animal tests to indicate that it is likely to be safe and efficacious when testing in humans. The typical data package required will include demonstrating compound safety, toxicity, bioavailability, pharmacodynamics (potential effects of the compound on body systems) and pharmacokinetics (how the compound is potentially absorbed, distributed, metabolised and eliminated after dosing in humans). If the desired criteria are met and the compound meets the clinical Candidate criteria and is deemed worthy of further development, a submission to regulatory bodies such as the US Food & Drug Administration for an exploratory Investigational New Drug Study can be made. The purpose of this study is to collect data to establish that the compound will not expose humans to unreasonable risks when used in limited, early-stage clinical studies in patients or normal volunteer subjects (Phase I). These studies are also designed to determine the metabolism and pharmacologic actions of the drug in humans, the side effects associated with increasing doses, and, if possible, to gain early evidence on their effectiveness. In order to reach the above goals, we have developed a pre-clinical high throughput Absorption, Distribution, Metabolism and Excretion–Toxicity (ADME–Toxicity) panel of assays to identify compounds that are likely to meet the Lead and Candidate compound acceptance criteria. This panel includes solubility studies in a range of biological fluids, cell viability studies in cancer and primary cell-lines, mitochondrial toxicity, off-target effects (across the kinase, protease, histone deacetylase, phosphodiesterase and GPCR protein families), CYP450 inhibition (5 different CYP450 enzymes), CYP450 induction, cardio-toxicity (hERG) and gene-toxicity. This panel of assays has been applied to multiple compound series developed in a number of projects delivering Lead and clinical Candidates and examples from these will be presented.

Keywords: absorption, distribution, metabolism and excretion–toxicity , drug discovery, food and drug administration , pharmacodynamics

Procedia PDF Downloads 173

Authors: Pomila Sharma

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The rapid urbanization in India was not accompanied by the establishment of waste water treatment facility at similar and same pace. The inland fresh water ecosystem is increasingly subjected to great stress from various human activities. Jalmahal Lake is located in Jaipur city of Rajasthan state; the lake was constructed about 400 years ago and surrounded by hills. The lake was approximately 139 hectare in full spread and has catchment area of 23.5 sq. kilometer. Out of the total catchment area approximate 40% falls inside dense urban area of Jaipur city. During the showers, the treated and untreated waste waters and runoff waters get mixed and enter the lake through the various influx channels, and the lake water quality gets affected by the inflow of waste water. The main objective of this work was to use the Moringa oleifera seeds as a natural adsorbent for the treatment of wastewater in lake. Moringa oleifera is a tropical, multipurpose tree whose seeds contain high-quality edible oil 40% by weight and water soluble, non-toxic protein that act as an effective coagulant for the removal of organic matter in water and waste water treatment. Laboratory Jar test procedure had been used for coagulation studies; an experiment runs using lake water. Water extracts/powder of Moringa seed applied to treat polluted water of lake. In present study various doses of Moringa oleifera seed coagulant viz. 100 mg/L, 200 mg/L, and 400 mg/L were taken and checked for the efficiency dose on treated and untreated polluted water. Turbidity and color removal is one of the important steps in a waste water treatment processes. The results indicate significant reduction in turbidity and color. Standard plate count was significantly reduced fecal coliform levels too. All parameters were reduced with the increased dose of Moringa oleifera. It was clear from the study Moringa oleifera seed was shown to be a potential bio-coagulant, for treatment of sewage laden polluted water in the lake.

Keywords: coagulant, Moringa oleifera, plate count, turbidity, wastewater

Procedia PDF Downloads 410

Authors: S. M. Ali, S. I. Mohamed

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Reduction of feed cost for broiler production is at most importance in decreasing the cost of production. The objectives of this study were to evaluate the use of synthetic amino acids (L-lysine – DL-methionine) instead of super concentrate and groundnut cake versus meat powder as protein sources. A total of 180 male broiler chicks (Cobb – strain) at 15 day of age (DOA) were selected according to their average body weight (380 g) from a broiler chicks flock at Elbashair Farm. The chicks were randomly divided into six groups of 30 chicks. Each group was further sub divided into three replicates with 10 birds. Six experimental diets were formulated. The first diet contained groundnut cake and super concentrate as the control (GNC + C); in the second diet, meat powder and super concentrate (MP + C) were used. The third diet contained groundnut cake and amino acids (GNC + AA); the forth diet contained meat powder and amino acids (MP + AA). The fifth diet contained groundnut cake, meat powder and super concentrate (GNC + MP + C) and the sixth diet contained groundnut cake, meat powder and amino acids (GNC + MP + AA). The formulated rations were randomly assigned for the different sub groups in a completely randomized design of six treatments and three replicates. Weekly feed intake, body weight and mortality were recorded and body weight gain and feed conversion ratio were calculated. At the end of the experiment (49 DOA), nine birds from each treatment were slaughtered. Live body weight, carcass weight, head, shank, and some internal organs (gizzard, heart, liver, small intestine, and abdominal fat pad) weights were taken. For the overall experimental period the (GNC + C +MP) consumed significantly (P≤0.01) the highest cumulative feed while the (MP + AA) group consumed the lowest amount of feed. The (GNC + C) and the (GNC + AA) groups had the heaviest live body weight while (MP + AA) had the lowest live body weight. The overall FCR was significantly (P≤0.01) the best for (GNC + AA) group while the (MP + AA) reported the worst FCR. However, the (GNC + AA) had significantly (P≤0.01) the lowest AFP. The (GNC + MP + Con) group had the highest dressing % while the (MP + AA) group had the lowest dressing %. It is concluded that amino acids can be used instead of super concentrate in broiler feeding with perfect performance and less cost and that meat powder is not advisable to be used with amino acids.

Keywords: broiler chickens, DL-lysine, methionine, performance

Procedia PDF Downloads 268

Authors: Vinod Kumar

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Background: Tissue Plasminogen Activator (tPA) showed a level 1 benefit in acute stroke (within 3-6 hrs). Intracarotid sodium nitroprusside (ICSNP) has been studied in this context with a wide treatment window, fast recovery and affordability. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a)Retrograde Neurotransmission (acute cases): 1)Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2)Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b)Vasospasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c)Long-Term Potentıatıon (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Des’gn: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat stroke cases. Case-control prospective study. Mater’als And Methods: The experimental population included 82 stroke patients (10 patients were given control treatments without superfusion or with 5% dextrose superfusion, and 72 patients comprised the ICSNP group). The mean time for superfusion was 9.5 days post-stroke. Pre- and post-ICSNP status was monitored by NIHSS, MRI and TCD. Results: After 90 seconds in the ICSNP group, the mean change in the NIHSS score was a decrease of 1.44 points, or 6.55%; after 2 h, there was a decrease of 1.16 points; after 24 h, there was an increase of 0.66 points, 2.25%, compared to the control-group increase of 0.7 points, or 3.53%; at 7 days, there was an 8.61-point decrease, 44.58%, compared to the control-group increase of 2.55 points, or 22.37%; at 2 months in ICSNP, there was a 6.94-points decrease, 62.80%, compared to the control-group decrease of 2.77 points, or 8.78%. TCD was documented and improvements were noted. Conclusions: ICSNP is a swift-acting drug in the treatment of stroke, acting within 90 seconds on day 9.5 post-stroke with a small decrease after 24 hours. The drug recovers from this decrease quickly.

Keywords: brain infarcts, intracarotid sodium nitroprusside, perforators, vasodilatıons, retrograde transmission, the 10, 000-fold effect

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Authors: T. Ul Rehman, S. Agnello, F. M. Gelardi, M. M. Calvino, G. Lazzara, G. Buscarino, M. Cannas

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Metal-Organic Frameworks (MOFs) have emerged as promising candidates for detecting metal ions owing to their large surface area, customizable porosity, and diverse functionalities. In recent years, there has been a surge in research focused on MOFs with luminescent properties. These frameworks are constructed through coordinated bonding between metal ions and multi-dentate ligands, resulting in inherent fluorescent structures. Their luminescent behavior is influenced by factors like structural composition, surface morphology, pore volume, and interactions with target analytes, particularly metal ions. MOFs exhibit various sensing mechanisms, including photo-induced electron transfer (PET) and charge transfer processes such as ligand-to-metal (LMCT) and metal-to-ligand (MLCT) transitions. Among these, MIL-53(Al) stands out due to its flexibility, stability, and specific affinity towards certain metal ions, making it a promising platform for selective metal ion sensing. This study investigates the structural, thermal, and luminescent properties of MIL-53(Al) metal-organic framework (MOF) upon Fe3+ cation exchange. Two separate sets of samples were prepared to activate the MOF powder at different temperatures. The first set of samples, referred to as MIL-53(Al), activated (120°C), was prepared by activating the raw powder in a glass tube at 120°C for 12 hours and then sealing it. The second set of samples, referred to as MIL-53(Al), activated (300°C), was prepared by activating the MIL-53(Al) powder in a glass tube at 300°C for 70 hours. Additionally, 25 mg of MIL-53(Al) powder was dispersed in 5 mL of Fe3+ solution at various concentrations (0.1-100 mM) for the cation exchange experiment. The suspension was centrifuged for five minutes at 10,000 rpm to extract MIL-53(Al) powder. After three rounds of washing with ultrapure water, MIL-53(Al) powder was heated at 120°C for 12 hours. For PXRD and TGA analyses, a sample of the obtained MIL-53(Al) was used. We also activated the cation-exchanged samples for time-resolved photoluminescence (TRPL) measurements at two distinct temperatures (120 and 300°C) for comparative analysis. Powder X-ray diffraction patterns reveal amorphization in samples with higher Fe3+ concentrations, attributed to alterations in coordination environments and ion exchange dynamics. Thermal decomposition analysis shows reduced weight loss in Fe3+-exchanged MOFs, indicating enhanced stability due to stronger metal-ligand bonds and altered decomposition pathways. Raman spectroscopy demonstrates intensity decrease, shape disruption, and frequency shifts, indicative of structural perturbations induced by cation exchange. Photoluminescence spectra exhibit ligand-based emission (π-π* or n-π*) and ligand-to-metal charge transfer (LMCT), influenced by activation temperature and Fe3+ incorporation. Quenching of luminescence intensity and shorter lifetimes upon Fe3+ exchange result from structural distortions and Fe3+ binding to organic linkers. In a nutshell, this research underscores the complex interplay between composition, structure, and properties in MOFs, offering insights into their potential for diverse applications in catalysis, gas storage, and luminescent devices.

Keywords: Fe³⁺ cation exchange, luminescent metal-organic frameworks (LMOFs), MIL-53(Al), solid-state analysis

Procedia PDF Downloads 66

Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy

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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.

Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin

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Authors: Virginia Martin Torrejon, Binjie Wu

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Gelatine is a protein biopolymer obtained from the partial hydrolysis of animal tissues which contain collagen, the primary structural component in connective tissue. Gelatine hydrogels have attracted considerable research in recent years as an alternative to synthetic materials due to their outstanding gelling properties, biocompatibility and compostability. Surfactants, such as sodium dodecyl sulfate (SDS), are often used in hydrogels solutions as surface modifiers or solubility enhancers, and their incorporation can influence the hydrogel’s viscoelastic properties and, in turn, its processing and applications. Literature usually focuses on studying the impact of formulation parameters (e.g., gelatine content, gelatine strength, additives incorporation) on gelatine hydrogels properties, but processing parameters, such as curing temperature, are commonly overlooked. For example, some authors have reported a decrease in gel strength at lower curing temperatures, but there is a lack of research on systematic viscoelastic characterisation of high strength gelatine and gelatine-SDS systems at a wide range of curing temperatures. This knowledge is essential to meet and adjust the technological requirements for different applications (e.g., viscosity, setting time, gel strength or melting/gelling temperature). This work investigated the effect of curing temperature (10, 15, 20, 23 and 25 and 30°C) on the elastic modulus (G’) and melting temperature of high strength gelatine-SDS hydrogels, at 10 wt% and 20 wt% gelatine contents, by small-amplitude oscillatory shear rheology coupled with Fourier Transform Infrared Spectroscopy. It also correlates the gel strength obtained by rheological measurements with the gel strength measured by texture analysis. Gelatine and gelatine-SDS hydrogels’ rheological behaviour strongly depended on the curing temperature, and its gel strength and melting temperature can be slightly modified to adjust it to given processing and applications needs. Lower curing temperatures led to gelatine and gelatine-SDS hydrogels with considerably higher storage modulus. However, their melting temperature was lower than those gels cured at higher temperatures and lower gel strength. This effect was more considerable at longer timescales. This behaviour is attributed to the development of thermal-resistant structures in the lower strength gels cured at higher temperatures.

Keywords: gelatine gelation kinetics, gelatine-SDS interactions, gelatine-surfactant hydrogels, melting and gelling temperature of gelatine gels, rheology of gelatine hydrogels

Procedia PDF Downloads 102

Authors: Hager E. Amer, Hany El Saftawy, Laila Rashed, Ahmed M. Ata, Fatma Metwally, Hesham Mettawei, Hossam E. Sayed, Tamer Adel, Kareem M. El Sawah

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Aim: The purpose of this study is to regenerate the damaged photoreceptor cells as a result of argon laser as a model of Retinitis Pigmentosa in rabbits' retina by using adult stem cells from rabbits' bone marrow. Background: Retinitis pigmentosa (RP) is a group of inherited disorders that primarily affect photoreceptor and pigment epithelium function. RP leads to loss of the rod outer segment and shorten the photoreceptor layer and expose the photoreceptor cell body to high-pressure levels in oxygen (oxidative stress) leads to apoptosis to the rod and cone cells. In particular, there is no specific treatment for retinitis pigmentosa. Materials and Methods: Forty Two Giant (Rex) rabbits were used in this experiment divided into 3 groups: Group 1: Control (6 rabbits), Group 2: Argon laser radiated as a model of retinitis pigmentosa (12 rabbits), Group 3: Laser radiated and treated by 6 million stem cells (12 rabbits). The last two groups are divided each into two subgroups each subgroup contains 6 rabbits, the ophthalmological examination was performed on rabbits, blood samples and retina samples were taken after 25 days and after 36 days from the laser radiation (10 days and 21 days after stem cells insertion in group 3) to perform the biochemical analysis. Results: Compared to control Group, a decrease of ERG wave amplitude and antioxidant substances (Glutathione) in blood and retina in group 2, and an increase of oxidative stress substances (Nitric oxide, Malonaldehyde, and carponyl protein) and apoptotic substances (Advanced glycation end product and M-metalloproteinase) in blood and retina. Compared to group 2, mostly increases of antioxidant substances and ERG wave amplitude in group 3, and mostly decreases in oxidative stress substances and apoptotic substances. Conclusion: Insertion of 6 million stem cells intravitreous gives good results in regeneration of the damaged photoreceptor cells after 21 days.

Keywords: retinitis pigmentosa, stem cells, argon laser, oxidative stress, apoptosis

Procedia PDF Downloads 199

Authors: Sunil Kumar, Uday C. Ghoshal

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Background and aims: Serotonin (5-hydroxtryptamine, 5-HT) is an important factor in gut function, playing key roles in intestinal peristalsis and secretion, and in sensory signaling in the brain-gut axis. Removal from its sites of action is mediated by a specific protein called the serotonin reuptake transporter (SERT). Polymorphisms in the promoter region of the SERT gene have effects on transcriptional activity, resulting in altered 5-HT reuptake efficiency. Functional polymorphisms may underlie disturbance in gut function in individuals suffering with disorders such as irritable bowel syndrome (IBS). The aim of this study was to assess the potential association between SERT polymorphisms and the diarrhea predominant IBS (D-IBS) phenotype Subjects: A total of 36 northern Indian female patients and 55 female northern Indian healthy controls (HC) were subjected to genotyping. Methods: Leucocyte DNA of all subjects was analyzed by polymerase chain reaction based technologies for SERT polymorphisms, specifically the insertion/deletion polymorphism in the promoter (SERT-P). Statistical analysis was performed to assess association of SERT polymorphism allele with the D-IBS phenotype. Results: The frequency of distribution of SERT-P gene was comparable between female patients with IBS and HC (p = 0.086). However, frequency of SERT-P deletion/deletion genotype was significantly higher in female patients with D-IBS compared to C-IBS and A-IBS [17/19 (89.5%) vs. 4/12 (33.3%) vs. 1/5 (20%), p=0.001, respectively]. The mucosal level of serotonin was higher in D-IBS compared to C-IBS and A-IBS [Median, range (159.26, 98.78–212.1) vs. 110.4, 67.87–143.53 vs. 92.34, 78.8–166.3 pmol/mL, p=0.001, respectively]. The mucosal level of serotonin was higher in female patients with IBS with SERT-P deletion/deletion genotype compared deletion/insertion and insertion/insertion [157.65, 67.87–212.1 vs. 110.4, 78.1–143.32 vs. 100.5, 69.1–132.03 pmol/mL, p=0.001, respectively]. Patients with D-IBS with deletion/deletion genotype more often reported symptoms of abdominal pain, discomfort (p=0.025) and bloating (p=0.039). Symptoms development following lactose ingestion was strongly associated with D-IBS and SERT-P deletion/deletion genotype (p=0.004). Conclusions: Significant association was observed between D-IBS and the SERT-P deletion/deletion genotype, suggesting that the serotonin transporter is a potential candidate gene for D-IBS in women.

Keywords: serotonin, SERT, inflammatory bowel disease, genetic polymorphism

Procedia PDF Downloads 333

Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu

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Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.

Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology

Procedia PDF Downloads 439

Authors: V. K. Tewari, M. Hussain, H. K. D. Gupta

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Background: Nitric Oxide Donars (NODs) (intrathecal sodium nitroprusside (ITSNP) and oral tadalafil 20mg post ITSNP) has been studied in this context in cerebral palsy patients for fast recovery. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a) Retrograde Neurotransmission (acute cases): 1) Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2) Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b) Vasopasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c) Long-Term Potentiation (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Design: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat cerebral palsy cases. Case-control prospective study. Materials and Methods: The experimental population included 82 cerebral palsy patients (10 patients were given control treatments without NOD or with 5% dextrose superfusion, and 72 patients comprised the NOD group). The mean time for superfusion was 5 months post-cerebral palsy. Pre- and post-NOD status was monitored by Gross Motor Function Classification System for Cerebral Palsy (GMFCS), MRI, and TCD studies. Results: After 7 days in the NOD group, the mean change in the GMFCS score was an increase of 1.2 points mean; after 3 months, there was an increase of 3.4 points mean, compared to the control-group increase of 0.1 points at 3 months. MRI and TCD documented the improvements. Conclusions: NOD (ITSNP boosts up the recovery and oral tadalafil maintains the recovery to a well-desired level) acts swiftly in the treatment of CP, acting within 7 days on 5 months post-cerebral palsy either of the three mechanisms.

Keywords: cerebral palsy, intrathecal sodium nitroprusside, oral tadalafil, perforators, vasodilations, retrograde transmission, the 10, 000-fold effect, long-term potantiation

Procedia PDF Downloads 364

Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

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NASH is an increasingly prevalent chronic liver disease that can progress to hepatocellular carcinoma and now is attracting interest worldwide. The STAM™ model is a clinically-correlated murine NASH model which shows the same pathological progression as NASH patients and has been widely used for pharmacological and basic research. The multiple parallel hits hypothesis suggests abnormalities in adipocytokines, intestinal microflora, and endotoxins are intertwined and could contribute to the development of NASH. In fact, NASH patients often exhibit gut dysbiosis and dysfunction in adipose tissue and metabolism. However, the analysis of the STAM™ model has only focused on the liver. To clarify whether the STAM™ model can also mimic multiple pathways of NASH progression, we analyzed the organ crosstalk interactions between the liver and the gut and the phenotype of adipose tissue in the STAM™ model. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin 2 days after birth and feeding with high-fat diet after 4 weeks of age. The mice were sacrificed at NASH stage. Colon samples were snap-frozen in liquid nitrogen and stored at -80˚C for tight junction-related protein analysis. Adipose tissue was prepared into paraffin blocks for HE staining. Blood adiponectin was analyzed to confirm changes in the adipocytokine profile. Tight junction-related proteins in the intestine showed that expression of ZO-1 decreased with the progression of the disease. Increased expression of endotoxin in the blood and decreased expression of Adiponectin were also observed. HE staining revealed hypertrophy of adipocytes. Decreased expression of ZO-1 in the intestine of STAM™ mice suggests the occurrence of leaky gut, and abnormalities in adipocytokine secretion were also observed. Together with the liver, phenotypes in these organs are highly similar to human NASH patients and might be involved in the pathogenesis of NASH.

Keywords: Non-alcoholic steatohepatitis, hepatocellular carcinoma, fibrosis, organ crosstalk, leaky gut

Procedia PDF Downloads 159

Authors: Ayhan Varol, Aysenur Ugurlu

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Due to the depletion of fossil fuels and climate change, there is a rising interest in renewable energy sources. In this concept, a wide range of biomass (energy crops, animal manure, solid wastes, etc.) are used for energy production. There has been a growing interest in biomethane production from biomass. Biomethane production from organic wastes is a promising alternative for waste management by providing organic matter stabilization. Anaerobic digestion of organic material produces biogas, and organic substrate is degraded into a more stable material. Therefore, anaerobic digestion technology helps reduction of carbon emissions and produces renewable energy. The hydraulic retention time (HRT) and organic loading rate (OLR), as well as TS (VS) loadings, influences the anaerobic digestion of organic wastes significantly. The optimum range for HRT varies between 15 days to 30 days, whereas OLR differs between 0.5 to 5 g/L.d depending on the substrate type and its lipid, protein and carbohydrate contents. The organic wastes have biogas production potential through anaerobic digestion. In this study, biomethane production potential of wastes like sugar beet bagasse, agricultural residues, food wastes, olive mill pulp, and dairy manure having different characteristics was investigated in mesophilic CSTR reactor, and their performances were compared. The reactor was mixed in order to provide homogenized content at a rate of 80 rpm. The organic matter content of these wastes was between 85 to 94 % with 61% (olive pulp) to 22 % (food waste) dry matter content. The hydraulic retention time changed between 20-30 days. High biogas productions, 13.45 to 5.70 mL/day, were achieved from the wastes studied when operated at 9 to 10.5% TS loadings where OLR varied between 2.92 and 3.95 gVS/L.day. The results showed that food wastes have higher specific methane production rate and volumetric methane production potential than the other wastes studied, under the similar OLR values. The SBP was 680, 585, 540, 390 and 295 mL/g VS for food waste, agricultural residues, sugar beet bagasse, olive pulp and dairy manure respectively. The methane content of the biogas varied between 72 and 60 %. The volatile solids conversion rate for food waste was 62%.

Keywords: biogas production, organic wastes, biomethane, anaerobic digestion

Procedia PDF Downloads 281

Authors: Mardiana Madjid, Nurpudji Astuti Taslim, Suryani As'ad, Haerani Rasyid, Agussalim Bukhari

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The high level of Urine Urea Nitrogen (UUN) indicates hypercatabolism occurs in hospitalized patients. High levels of Total Lymphocyte Count (TLC) indicates the immune system condition, adequate wound healing, and limit complication. Adequate dietary intake affects to decrease of hypercatabolism status in treated patient’s hospitals. Nitrogen Balance (NB) is simply the difference between nitrogen (N₂) intake and output. If more N₂ intake than output, then positive NB or anabolic will occur. This study aims to evaluate the effect of dietary intake in influencing balance nitrogen and total lymphocyte count. Method: A total of 43 patients admitted to a Wahidin Sudirohusodo Hospital between 2018 and 2019 for 10 days' treats are included. The inclusion criteria were patients who were treated for 10 days and receives food from the hospital orally. Patients did not experience gastrointestinal disorders such as vomiting and diarrhea and experience impair kidney function and liver function and expressed approval to participate in this study. During hospitalization, food intake, UUN, albumin serum, balance nitrogen, and TLC was assessed twice on day 1 and day 10. There is no Physician Clinical Nutritional intervention to correct food intake. UUN is 24 hours of urine collected on the second day after admission and the tenth day. Statistical analysis uses SPSS 24 with observational cohort methods. Result: The Forty-three participants completed the follow-up (27 men and 18 women). The age of fewer than 4 years is 22 people, 45 to 60 years is 16 people, and over 60 years is 4 people. The result of the study on day 1 obtained SGA score A, SGA score B, SGA score C are 8, 32, 3 until day 10 are 8, 31, 4, respectively. According to 24h dietary recalls, the energy intake during observation was from 522.5 ± 400.4 to 1011.9 ± 545.1 kcal/day P < 0.05, protein intake from 20.07 ± 17.2 to 40.3 ± 27.3 g/day P < 0.05, carbohydrates from 92.5 ± 71.6 to 184.8 ± 87.4 g/day, and fat from 5.5 ± 3.86 to 13.9 ± 13.9 g/day. The UUN during the observation was from 6.6 ± 7.3 to 5.5 ± 3.9 g/day, TLC decreased from 1622.9 ± 897.2 to 1319.9 ± 636.3/mm³ value target 1800/mm³, albumin serum from 3.07 ± 0.76 to 2.9 ± 0.57 g/day, and BN from -7.5 ± 7.2 to -3.1 ± 4.86. Conclusion: The high level of UUN needs to correct adequate dietary intake to improve NB and TLC status on hospitalized patients.

Keywords: adequate dietary intake, balance nitrogen, total lymphocyte count, urine urea nitrogen

Procedia PDF Downloads 127

Authors: Hamza Javar Magnier, Leslie V. Woodcock

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We report extensive molecular dynamics (MD) computational investigations into the thermodynamic description of supercritical properties for a model fluid that is the simplest realistic representation of atoms or molecules. The pair potential is a hard-sphere repulsion of diameter σ with a very short attraction of length λσ. When λ = 1.005 the range is so short that the model atoms are referred to as “adhesive spheres”. Molecular dimers, trimers …etc. up to large clusters, or droplets, of many adhesive-sphere atoms are unambiguously defined. This then defines percolation transitions at the molecular level that bound the existence of gas and liquid phases at supercritical temperatures, and which define the existence of a supercritical mesophase. Both liquid and gas phases are seen to terminate at the loci of percolation transitions, and below a second characteristic temperature (Tc2) are separated by the supercritical mesophase. An analysis of the distribution of clusters in gas, meso- and liquid phases confirms the colloidal nature of this mesophase. The general phase behaviour is compared with both experimental properties of the water-steam supercritical region and also with formally exact cluster theory of Mayer and Mayer. Both are found to be consistent with the present findings that in this system the supercritical mesophase narrows in density with increasing T > Tc and terminates at a higher Tc2 at a confluence of the primary percolation loci. The expended plot of the MD data points in the mesophase of 7 critical and supercritical isotherms in highlight this narrowing in density of the linear-slope region of the mesophase as temperature is increased above the critical. This linearity in the mesophase implies the existence of a linear combination rule between gas and liquid which is an extension of the Lever rule in the subcritical region, and can be used to obtain critical parameters without resorting to experimental data in the two-phase region. Using this combination rule, the calculated critical parameters Tc = 0.2007 and Pc = 0.0278 are found be agree with the values found by of Largo and coworkers. The properties of this supercritical mesophase are shown to be consistent with an alternative description of the phenomenon of critical opalescence seen in the supercritical region of both molecular and colloidal-protein supercritical fluids.

Keywords: critical opalescence, supercritical, square-well, percolation transition, critical parameters.

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Authors: May Phyo Wai, Tanyawan Suantawee

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The positive correlation between unhealthy diets (high in fats, sugars, carbohydrates, and low fibers) and the risk of non-communicable diseases (NCDs) like obesity, hypertension, diabetes, and heart diseases has led to a growing interest in healthier lifestyles and diets. Consequently, people are opting for foods rich in fiber and phytochemicals. Wolffia globosa, also known as duckweed or watermeal, is the smallest plant with high nutritional value, including protein, fiber, phytochemicals, and antioxidant properties. It offers numerous health benefits, such as improving gut health and lowering blood glucose levels, and it is widely available in Thailand. The purpose of this study was to develop nutritionally enhanced breadsticks utilizing vacuum heat-dried Wolffia globosa power (WP). Various concentrations of WP (0% as control, 5%, 10%, and 15 % w/w/) were added, and then the breadsticks’ physical properties (hardness, fracturability, and color), phytochemicals (total phenolic compounds: TPC and total flavonoid contents: TFC), and antioxidant properties (DPPH radical scavenging activity (DPPH) and ferric reducing antioxidant power (FRAP) assay) were investigated. Experiments were done by triplicates and data was analyzed by one-way ANOVA. The results showed that the hardness, measured by a texture analyzer, increased significantly (p<0.05) with higher WP concentrations, reaching 2,897.01 ± 77.31 g at 15% WP from 1,314.41 ± 32.52 g of the control. In contrast, the lightness (L*), redness (a*), and yellowness (b*) of the breadsticks significantly decreased (p < 0.05) in a dose-dependent manner with added WP. Incorporating WP, rich in phytochemicals and antioxidants, into the flour significantly enhanced the TPC and TFC of the breadsticks (p<0.05), with TPC and TFC increasing dose-dependently rising to 1.8-fold and 3.5-fold at 15% WP, respectively. The antioxidant power, assessed by DPPH and FRAP assays, also showed a similar trend, with significantly higher values at 10% and 15% WP (p<0.05). These results indicate that adding WP significantly boosted the TPC, TFC, DPPH, and FRAP values of the developed breadsticks. Therefore, incorporating WP into breadsticks might be a promising strategy for creating food products enriched with phytochemicals and antioxidants, offering consumers healthier options in the market.

Keywords: antioxidant properties, breadsticks, phytochemicals, Wolffia globosa

Procedia PDF Downloads 37

Authors: Rossy G. Olan Villegas, Gerardo Acosta Garcia, Aurea Bernardino Nicanor, Leopoldo Gonzalez Cruz, Humberto Ramirez Medina

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Capsicum annuum is a crop of agricultural and economic importance in Mexico and other countries. The fruit (pepper) contains bioactive components such as carotenoids, phenolic compounds and capsaicinoids that improve health. However, pepper cultivation is affected by biotic and abiotic factors that decrease yield. Some phytohormones like gibberellins and auxins induce the formation and development of fruit in several plants. In this study, we evaluated the effect of the exogenous application of phytohormones like gibberellic acid and indolbutyric acid on fruit development of jalapeno pepper plants, the protein profile of plant tissues, the accumulation of bioactive compounds and antioxidant activity in the pericarp and seeds. For that, plants were sprinkled with these phytohormones. The fruit collection for the control, indolbutyric acid and gibberellic acid treatments was 7 peppers per plant; however, for the treatment that combines indolbutyric acid and gibberellic acid, a fruit with the shortest length (1.52 ± 1.00 cm) and weight (0.41 ± 1.0 g) was collected compared to fruits of plants grown under other treatments. The length (4,179 ± 0,130 cm) and weight of the fruit (8,949 ± 0.583 g) increased in plants treated with indolbutyric acid, but these characteristics decreased with the application of GA3 (length of 3,349 ± 0.127 cm and a weight 4,429 ± 0.144 g). The content of carotenes and phenolic compounds increased in plants treated with GA3 (1,733 ± 0.092 and 1,449 ± 0.009 mg / g, respectively) or indolbutyric acid (1,164 ± 0.042 and 0.970 ± 0.003 mg / g). However, this effect was not observed in plants treated with both phytohormones (0.238 ± 0.021 and 0.218 ± 0.004 mg / g). Capsaicin content was higher in all treatments; but it was more noticeable in plants treated with both phytohormones, the value being 0.913 ± 0.001 mg / g (three times greater in amount). The antioxidant activity was measured by 3 different assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH), antioxidant power of ferric reduction (FRAP) and 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid ( ABTS) to find the minimum inhibitory concentration of the reducing radical (IC50 and EC50). Significant differences were observed from the application of the phytohormone, being the fruits treated with gibberellins, which had a greater accumulation of bioactive compounds. Our results suggest that the application of phytohormones modifies the development of fruit and its content of bioactive compounds.

Keywords: auxins, capsaicinoids, carotenoids, gibberellins

Procedia PDF Downloads 116

Authors: Alireza Barari, Saeed Shirali, Ahmad Abdi

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Abstract: Introduction: Physical activity may be related to male reproductive function by affecting on thehypothalamic-pituitary-gonadal(HPG) axis. Our aim was to determine the effects of 6 weeks resistance exercise on reproductive hormones, HPG axis. The hypothalamic-pituitary-gonadal (HPG) axis refers tothe effects of endocrine glands in three-level including (i) the hypothalamic releasing hormone GnRH, which is synthesized in in a small heterogenous neuronal population and released in a pulsatile fashion, (ii) the anterior pituitary hormones, follicle-stimulating hormone(FSH) and luteinizing hormone (LH) and (iii) the gonadal hormones, which include both steroid such as testosterone (T), estradiol and progesterone and peptide hormones (such as inhibin). Hormonal changes that create a more anabolic environment have been suggested to contribute to the adaptation to strength exercise. Physical activity has an extensive impact on male reproductive function depending upon the intensity and duration of the exercise and the fitness level of the individual. However, strenuous exercise represents a physical stress and inflammation changed that challenges homeostasis. Materials and methods: Sixteen male volunteered were included in a 6-week control period followed by 6 weeks of resistance training (leg press, lat pull, chest press, squat, seatedrow, abdominal crunch, shoulder press, biceps curl and triceps press down) four times per week. intensity of training loading was 60%-75% of one maximum repetition. Participants performed 3 sets of 10 repetitions. Rest periods were two min between exercises and sets. Start with warm up exercises include: The muscles relax and stretch the body, which was for 10 minutes. Body composition, VO2max and the circulating level of free testosterone (fT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), sex hormone binding globulin (SHBG) and inhibin B measured prior and post 6-week intervention. The hormonal levels of each serum sample were measured using commercially available ELISA kits. Analysis of anthropometrical data and hormonal level were compared using the independent samples t- test in both groups and using SPSS (version 19). P ≤ 0.05 was considered statistically significant. Results: For muscle strength, both lower- and upper-body strength were increased significantly. Aerobic fitness level improved in trained participant from 39.4 ± 5.6 to 41.9 ± 5.3 (P = 0.002). fT concentration rise progressively in the trained group and was significantly greater than those in the control group (P = 0.000). By the end of the 6-week resistance training, serum SHBG significantly increased in the trained group compared with the control group (P = 0.013). In response to resistance training, LH, FSH and inhibin B were not significantly changed. Discussion: According to our finfings, 6 weeks of resistance training induce fat loss without any changes in body weight and BMI. A decline of 25.3% in percentage of body fat with statiscally same weight was due to increase in muscle mass that happened during resistance exercise periods . Six weeks of resistance training resulted in significant improvement in BF%, VO2max and increasing strength and the level of fT and SHBG.

Keywords: resistance, hypothalamic, pituitary, gonadal axis

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Authors: Min Liu, Jirui Gong, Qinpu Luo, Lili Yang, Bo Yang, Zihe Zhang, Yan Pan, Zhanwei Zhai

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Grazing disturbance is one of the important land-use types that affect plant growth and ecosystem processes. In order to study the responses of dominant species to grazing in the semiarid temperate grassland of Inner Mongolia, we set five grazing intensity plots: a control and four levels of grazing (light (LG), moderate (MG), heavy (HG) and extreme heavy grazing (EHG)) to test the morphological and physiological responses of Stipa grandis, Leymus chinensis at the individual levels. With the increase of grazing intensity, Stipa grandis and Leymus chinensis both exhibited reduced plant height, leaf area, stem length and aboveground biomass, showing a significant dwarf phenomenon especially in HG and EHG plots. The photosynthetic capacity decreased along the grazing gradient. Especially in the MG plot, the two dominant species have lowest net photosynthetic rate (Pn) and water use efficiency (WUE). However, in the HG and EHG plots, the two species had high light saturation point (LSP) and low light compensation point (LCP), indicating they have high light-use efficiency. They showed a stimulation of compensatory photosynthesis to the remnant leaves as compared with grasses in MG plot. For Leymus chinensis, the lipid peroxidation level did not increase with the low malondialdehyde (MDA) content even in the EHG plot. It may be due to the high enzymes activity of superoxide dismutase (SOD) and peroxidase (POD) to reduce the damage of reactive oxygen species. Meanwhile, more carbohydrate was stored in the leaf of Leymus chinensis to provide energy to the plant regrowth. On the contrary, Stipa grandis showed the high level of lipid peroxidation especially in the HG and EHG plots with decreased antioxidant enzymes activity. The soluble protein content did not change significantly in the different plots. Therefore, with the increase of grazing intensity, plants changed morphological and physiological traits to defend themselves effectively to herbivores. Leymus chinensis is more resistant to grazing than Stipa grandis in terms of tolerance traits, particularly under heavy grazing pressure.

Keywords: antioxidant enzymes activity, grazing density, morphological responses, photosynthesis

Procedia PDF Downloads 367

Authors: O. A. Adebiyi, A. O. Oni, A. O. K. Adeshehinwa, I. O. Adejumo

Abstract:

In forty nine (49) days, twenty four (24) growing pigs (Landrace x Large white) with an average weight of 17 ±2.1kg were allocated to four experimental treatments T1 (dry mash without probiotics), T2 (wet feed without probiotics), T3 (dry mash + Saccaromyces cereviseae probiotics) and T4 (wet feed + Saccaromyces cereviseae probiotics) which were replicated three times with two pigs per replicate in a completely randomised design. The basal feed (dry feed) was formulated to meet the nutritional requirement of the animal with crude protein of 18.00% and metabolisable energy of 2784.00kcal/kgME. Growth pattern, faecal microbial load and behavioural activities (eating, drinking, physical pen interaction and frequency of visiting the drinking troughs) were accessed. Pigs fed dry mash without probiotics (T1) had the highest daily feed intake among the experimental animals (1.10kg) while pigs on supplemented diets (T3 and T4) had an average daily feed intake of 0.95kg. However, the feed conversion ratio was significantly (p < 0.05) affected with pigs on T3 having least value of 6.26 compared those on T4 (wet feed + Saccaromyces cereviseae) with means of 7.41. Total organism counts varied significantly (p < 0.05) with pigs on T1, T2, T3 and T4 with mean values of 179.50 x106cfu; 132.00 x 106cfu; 32.00 x 106cfu and 64.50 x 106cfu respectively. Coliform count was also significantly (p < 0.05) different among the treatments with corresponding values of 117.50 x 106cfu; 49.00 x 106cfu, 8.00 x 106cfu for pigs in T1, T2 and T4 respectively. The faecal Saccaromyces cereviseae was significantly lower in pigs fed supplemented diets compared to their counterparts on unsupplemented diets. This could be due to the inability of yeast organisms to be voided easily through feaces. The pigs in T1 spent the most time eating (7.88%) while their counterparts on T3 spent the least time eating. The corresponding physical pen interaction times expressed in percentage of a day for pigs in T1, T2, T3 and T4 are 6.22%, 5.92%, 4.04% and 4.80% respectively. These behavioural responses exhibited by these pigs (T3) showed that little amount of dry feed supplemented with probiotics is needed for better performance. The water intake increases as a result of the dryness of the feed with consequent decrease in pen interaction and more time was spent resting than engaging in other possible vice-habit like fighting or tail biting. Pigs fed dry feed (T3) which was supplemented with Saccaromyces cereviseae probiotics had a better overall performance, least faecal microbial load than wet fed pigs either supplemented with Saccaromyces cereviseae or non-supplemented.

Keywords: behaviour, feed forms, feed utilization, growth, microbial

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Authors: Ram Sharma, Esha Chatterjee, Santosh Kumar Guru, Kunal Nepali

Abstract:

A tractable anti-lung cancer agent was identified via the installation of a Ring C expanded synthetic analogue of the alkaloid vasicinone [7,8,9,10-tetrahydroazepino[2,1-b] quinazolin-12(6H)-one (TAZQ)] as a surface recognition part in the HDAC inhibitory three-component model. Noteworthy to mention that the candidature of TAZQ was deemed suitable for accommodation in HDAC inhibitory pharmacophore as per the results of the fragment recruitment process conducted by our laboratory. TAZQ was pinpointed through the fragment screening program as a synthetically flexible fragment endowed with some moderate cell growth inhibitory activity against the lung cancer cell lines, and it was anticipated that the use of the aforementioned fragment to generate hydroxamic acid functionality (zinc-binding motif) bearing HDAC inhibitors would boost the antitumor efficacy of TAZQ. Consistent with our aim of applying epigenetic targets to the treatment of lung cancer, a strikingly potent anti-lung cancer scaffold (compound 6) was pinpointed through a series of in-vitro experiments. Notably, the compounds manifested a magnificent activity profile against KRAS and EGFR mutant lung cancer cell lines (IC50 = 0.80 - 0.96 µM), and the effects were found to be mediated through preferential HDAC6 inhibition (IC50 = 12.9 nM). In addition to HDAC6 inhibition, the compounds also elicited HDAC1 and HDAC3 inhibitory activity with an IC50 value of 49.9 nM and 68.5 nM, respectively. The HDAC inhibitory ability of compound 6 was also confirmed from the results of the western blot experiment that revealed its potential to decrease the expression levels of HDAC isoforms (HDAC1, HDAC3, and HDAC6). Noteworthy to mention that complete downregulation of the HDAC6 isoform was exerted by compound 6 at 0.5 and 1 µM. Moreover, in another western blot experiment, treatment with hydroxamic acid 6 led to upregulation of H3 acK9 and α-Tubulin acK40 levels, ascertaining its inhibitory activity toward both the class I HDACs and Class II B HDACs. The results of other assays were also encouraging as treatment with compound 6 led to the suppression of the colony formation ability of A549 cells, induction of apoptosis, and increase in autophagic flux. In silico studies led us to rationalize the results of the experimental assay, and some key interactions of compound 6 with the amino acid residues of HDAC isoforms were identified. In light of the impressive activity spectrum of compound 6, a pH-responsive nanocarrier (hyaluronic acid-compound 6 nanoparticles) was prepared. The dialysis bag approach was used for the assessment of the nanoparticles under both normal and acidic circumstances, and the pH-sensitive nature of hyaluronic acid-compound 6 nanoparticles was confirmed. Delightfully, the nanoformulation was devoid of cytotoxicity against the L929 mouse fibroblast cells (normal settings) and exhibited selective cytotoxicity towards the A549 lung cancer cell lines. In a nutshell, compound 6 appears to be a promising adduct, and a detailed investigation of this compound might yield a therapeutic for the treatment of lung cancer.

Keywords: HDAC inhibitors, lung cancer, scaffold, hyaluronic acid, nanoparticles

Procedia PDF Downloads 95

Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

Abstract:

Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

Procedia PDF Downloads 143

Authors: Manal Elsayed Elkheshin, Rasha Saleh Mansour, Mohamed Fawzy Mohamed Eldnasury, Mamdouh Elbadry Mohamed

Abstract:

This research Aims to identify the main factors affecting the production and the fish consumption in Egypt, through the econometric estimation for various forms functions of fish production and fish consumption during the period (1991-2014), as the aim of this research to forecast the production and the fish consumption in Egypt until 2020, through determine the best standard methods using (ARIMA).This research also aims to the economic feasibility of the production of fish in aquaculture farms study; investment cost and represents the value of land, buildings, equipment and irrigation. Aquaculture requires three types of fish (Tilapia, carp fish, and mullet fish), and the total area of the farm, about an acre. The annual Fish production from this project about 3.5 tons. The annual investment costs of about 50500 pounds, Find conclude that the project can repay the cost of their investments after about 4 years and 5 months, and therefore recommend the implementation of the project, and internal rate of return reached (IRR) of about 22.1%, where it is clear that the rate of large internal rate of return, and achieves pound invested in this project annual return is estimated at 22.1 pounds, more than the opportunity cost, so we recommend the need to implement the project.Recommendations:1. Increasing the fish agriculture to decrease the gap of animal protein. 2.Increasing the number of mechanism fishing boats, and the provision of transport equipped to maintain the quality of fish production. 3.Encourage and attract the local and foreign investments, providing advice to the investor on the aquaculture field. 4. Action newsletters awareness of the importance of these projects where these projects resulted in a net profit after recovery in less than five years, IRR amounted to about 23%, which is much more than the opportunity cost of a bank interest rate is about 7%, helping to create work and graduates opportunities, and contribute to the reduction of imports of the fish, and improve the performance of the food trade balance.

Keywords: equation model, individual share, red meat, consumption, production, endogenous variable, exogenous variable, financial performance evaluates fish culture, feasibility study, fish production, aquaculture

Procedia PDF Downloads 370