Search results for: milk proteins

Authors: Summer Hassan, David Crossman

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Myocardial fibrosis (MF) has been loosely defined as the process occurring in the pathological remodeling of the myocardium due to excessive production and deposition of extracellular matrix (ECM) proteins, including collagen. This reduces tissue compliance and accelerates progression to heart failure, as well as affecting the electrical properties of the myocytes resulting in arrhythmias. Microscopic interrogation of MF is key to understanding the molecular orchestrators of disease. It is well-established that recruitment and stimulation of myofibroblasts result in Collagen deposition and the resulting expansion in the ECM. Many types of Collagens have been identified and implicated in scarring of tissue. In a series of experiments conducted at our lab, we aim to elucidate the role collagen VI plays in the development of myocardial fibrosis and its direct impact on myocardial function. This was investigated through an animal experiment in Rats with Collagen VI knockout diseased and healthy animals as well as Collagen VI wild diseased and healthy rats. Echocardiogram assessments of these rats ensued at four-time points, followed by microscopic interrogation of the myocardium aiming to correlate the role collagen VI plays in myocardial function. Our results demonstrate a deterioration in cardiac function as represented by the ejection fraction in the knockout healthy and diseased rats. This elucidates a potential protective role that collagen-VI plays following a myocardial insult. Current work is dedicated to the microscopic characterisation of the fibrotic process in all rat groups, with the results to follow.

Keywords: heart failure, myocardial fibrosis, collagen, echocardiogram, confocal microscopy

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Authors: Djeukeu Asongni William, Leng Marlyse, Gouado Inocent

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Background: Process for obtaining D. schimperiana chips involves a long period of cooking followed by drying of obtained products in the sun. Such a process could induce the modification of the functional properties of the chips, thus reducing the technological uses of these products. This study was conducted with a view to assessing the impact of this process on the chips of D. schimperiana. Methods: The chips used were purchased in Baham, Bamendjou and Bagangté markets during the month of February 2013. A representative sample of each market chips was formed by mixing the chips of several sellers. The control sample consisted of fresh yams that have been sliced to the average size of local chips then dried in the oven at 45 ° C for 36 h. On each sample was performed the analysis of the physico-chemical properties (carbohydrates, lipids, proteins, iron , phosphorus, reducing sugars, ash and total starch) and gelling properties both with and without inhibitor alpha-amylases (0.018 and 0.146 mol / l). Results: Results show that the levels of ash 2.99 g / 100gms, iron 1.01 g / 100gms and phosphorus 532.06 mg / 100gms fresh sample were significantly higher than those of the products obtained in the traditional process. The functional properties of the chips obtained from different methods shows that the peak viscosity of the fresh sample is larger than the other samples with or without inhibitor. In addition, the fresh sample has the lowest breakdown under the same conditions. Conclusion: These results show that traditional process reduces technological potential of chips, thus limiting the value of D. schimperiana.

Keywords: Dioscorea schimperiana, chips, functional properties, technological properties, valorization

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Authors: Chia-Jung Li, Li-Te Lin, Kuan-Hao Tsui

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The study identifies SPP1 as a potential gene associated with ovarian aging, revealing a significant decline in its expression in aged ovaries. SPP1, also known as osteopontin (OPN), is a multifunctional glycoprotein involved with regulatory proteins and pro-inflammatory immune chemokines. However, its genetic links to ovarian aging have not been extensively explored. Spatial transcriptomic analyses were conducted on ovaries from young and aged female mice, along with a sample from a 73-year-old individual. Additionally, single-cell RNA sequencing analysis was performed to identify associations between SPP1 and key genes. The study focused on crucial genes, including ITGAV, ITGB1, CD44, MMP3, and FN1, with a particular emphasis on the correlation between SPP1 and ITGB1. The findings indicate a significant decline in SPP1 expression in aged ovaries, which was consistent in the 73-year-old sample. Single-cell RNA sequencing unveiled associations between SPP1 and key genes, emphasizing a strong co-expression correlation between SPP1 and ITGB1. While the study provides valuable insights, further research is necessary to understand the broader implications and potential applications of SPP1 in ovarian aging. Translating these findings to clinical settings requires careful consideration. The identification of SPP1 as a gene implicated in ovarian aging opens new avenues for advancing precision medicine and refining treatment strategies for conditions related to ovarian aging.

Keywords: SPP1, ovarian aging, spatial transcriptomic, single-cell RNA sequencing

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Authors: Tsai-chun Lai, Szu-ju Fu, Tzu-lin Lee, Yuh-Lien Chen

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There is much evidence that exposure to fine particulate matter (PM) from air pollution increases the risk of cardiovascular morbidity and mortality. According to previous reports, PM in the air enters the respiratory tract, contacts the alveoli, and enters the blood circulation, leading to the progression of cardiovascular disease. PM pollution may also lead to cardiometabolic disturbances, increasing the risk of cardiovascular disease. The effects of PM on cardiac function and mitochondrial damage are currently unknown. We used mice and rat cardiomyocytes (H9c2) as animal and in vitro cell models, respectively, to simulate an air pollution environment using PM. These results indicate that the apoptosis-related factor PUMA, a regulator of apoptosis upregulated by p53, is increased in mice treated with PM. Apoptosis was aggravated in cardiomyocytes treated with PM, as measured by TUNEL assay and Annexin V/PI. Western blot results showed that CASPASE3 was significantly increased and BCL2 (B-cell lymphoid 2) was significantly decreased under PM treatment. Concurrent exposure to PM increases mitochondrial reactive oxygen species (ROS) production by MitoSOX Red staining. Furthermore, using Mitotracker staining, PM treatment significantly shortened mitochondrial length, indicating mitochondrial fission. The expression of mitochondrial fission-related proteins p-DRP1 (phosphodynamics-related protein 1) and FIS1 (mitochondrial fission 1 protein) was significantly increased. Based on these results, the exposure to PM worsens mitochondrial function and leads to cardiomyocyte apoptosis.

Keywords: particulate matter, cardiomyocyte, apoptosis, mitochondria

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Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

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Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

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Authors: Rohit Singh Dangi, Ravi Kant Pal, Monica Sundd

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Acyl-CoA binding protein (ACBP) is a housekeeping protein which functions as an intracellular carrier of acyl-CoA esters. Given the fact that the amastigote stage (blood stage) of Leishmania depends largely on fatty acids as the energy source, of which a large part is derived from its host, these proteins might have an important role in its survival. In Leishmania major, genome sequencing suggests the presence of six ACBPs, whose function remains largely unknown. For functional and structural characterization, one of the ACBP genes was cloned, and the protein was expressed and purified heterologously. Acyl-CoA ester binding and stoichiometry were analyzed by isothermal titration calorimetry and Dynamic light scattering. Our results shed light on high affinity of ACBP towards longer acyl-CoA esters, such as myristoyl-CoA to arachidonoyl-CoA with single binding site. To understand the binding mechanism & dynamics, Nuclear magnetic resonance assignments of this protein are being done. The protein's crystal structure was determined at 1.5Å resolution and revealed a classical topology for ACBP, containing four alpha-helical bundles. In the binding pocket, the loop between the first and the second helix (16 – 26AA) is four residues longer from other extensively studied ACBPs (PfACBP) and it curls upwards towards the pantothenate moiety of CoA to provide a large tunnel space for long acyl chain insertion.

Keywords: acyl-coa binding protein (ACBP), acyl-coa esters, crystal structure, isothermal titration, calorimetry, Leishmania

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Authors: Cristina Cirone, Katrina Cirone, Monali S. Malvankar-Mehta

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Purpose: An increased risk of visual impairment has been observed in individuals lacking a balanced diet. The purpose of this paper is to characterize the relationship between plant-based diets and specific ocular outcomes among adults. Design: Systematic review and meta-analysis. Methods: This systematic review and meta-analysis were conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement guidelines. The databases MEDLINE, EMBASE, Cochrane, and PubMed, were systematically searched up until May 27, 2021. Of the 503 articles independently screened by two reviewers, 21 were included in this review. Quality assessment and data extraction were performed by both reviewers. Meta-analysis was conducted using STATA 15.0. Fixed-effect and random-effect models were computed based on heterogeneity. Results: A total of 503 studies were identified which then underwent duplicate removal and a title and abstract screen. The remaining 61 studies underwent a full-text screen, 21 progressed to data extraction and fifteen were included in the quantitative analysis. Meta-analysis indicated that regular consumption of fish (OR = 0.70; CI: [0.62-0.79]) and skim milk, poultry, and non-meat animal products (OR = 0.70; CI: [0.61-0.79]) is positively correlated with a reduced risk of visual impairment (age-related macular degeneration, age-related maculopathy, cataract development, and central geographic atrophy) among adults. Consumption of red meat [OR = 1.41; CI: [1.07-1.86]) is associated with an increased risk of visual impairment. Conclusion: Overall, a pescatarian diet is associated with the most favorable visual outcomes among adults, while the consumption of red meat appears to negatively impact vision. Results suggest a need for more local and government-led interventions promoting a healthy and balanced diet.

Keywords: plant-based diet, pescatarian diet, visual impairment, systematic review, meta-analysis

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Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

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Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

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Authors: Ceyda Dadali, Yeşim Elmaci

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Coffee was introduced as an economic crop during the fifteenth century; nowadays it is the most important food commodity ranking second after crude oil. Desirable sensory properties make coffee one of the most often consumed and most popular beverages in the world. The coffee preparation method has a significant effect on flavor and composition of coffee brews. Three different extraction methodologies namely decoction, infusion and pressure methods have been used for coffee brew preparation. Each of these methods is related to specific granulation (coffee grind) of coffee powder, water-coffee ratio temperature and brewing time. Coffee is a mixture of 1500 chemical compounds. Chemical composition of coffee highly depends on brewing methods, coffee bean species and roasting time-temperature. Coffee contains a wide number of very important bioactive compounds, such as diterpenes: cafestol and kahweol, alkaloids: caffeine, theobromine and trigonelline, melanoidins, phenolic compounds. The phenolic compounds of coffee include chlorogenic acids (quinyl esters of hidroxycinnamic acids), caffeic, ferulic, p-coumaric acid. In coffee caffeoylquinic acids, feruloylquinic acids and di-caffeoylquinic acids are three main groups of chlorogenic acids constitues 6% -10% of dry weight of coffee. The bioavailability of chlorogenic acids in coffee depends on the absorption and metabolization to biomarkers in individuals. Also, the interaction of coffee polyphenols with other compounds such as dietary proteins affects the biomarkers. Since bioactive composition of coffee depends on brewing methods effect of coffee brewing method on bioactive compounds of coffee will be discussed in this study.

Keywords: bioactive compounds of coffee, biomarkers, coffee brew, effect of brewing

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Authors: Ruyuf Y. Alnafisah, Nouf S. Alammari, Amani S. Alqahtani

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Background: Beverage choice can have implications for the risk of non-communicable diseases. However, there is a lack of knowledge in assessing the nutritional content of these beverages. This study aims to describe the nutrient content of pre-packaged beverages available in the Saudi market. Design: Data were collected from the Saudi Branded Food Data-base (SBFD). Nutrient content was standardized in terms of units and reference volumes to ensure consistency in analysis. Results: A total of 1490 beverages were analyzed. The highest median levels of the majority of nutrients were found among dairy products; energy (68.4(43-188]kcal/100 ml in a milkshake); protein (8.2(0.5-8.2]g/100 ml in yogurt drinks); total fat (2.1(1.3-3.5]g/100 ml in milk); saturated fat (1.4(0-1.4]g/100 ml in yogurt drinks); cholesterol (30(0-30]mg/100 ml in yogurt drinks); sodium (65(65-65].4mg/100 ml in yogurt drinks); and total sugars (12.9(7.5-27]g/100 ml in milkshake). Carbohydrate level was the highest in nectar (13(11.8-14.2] g/100ml]; fruits drinks (12.9(11.9-13.9] g/100ml), and sparkling juices (12.9(8.8-14] g/100ml). The highest added sugar level was observed among regular soft drinks (12(10.8-14] g/100ml). The average rate of nutrient declaration was 60.95%. Carbo-hydrate had the highest declaration rate among nutrients (99.1%), and yogurt drinks had the highest declaration rate among beverage categories (92.7%). The median content of vitamins A and D in dairy products met the mandatory addition levels. Conclusion: This study provides valuable insights into the nutrient content of pre-packaged beverages in the Saudi market. It serves as a foundation for future research and monitoring. The findings of the study support the idea of taxing sugary beverages and raise concerns about the health effects of high sugar in fruit juices. Despite the inclusion of vitamins D and A in dairy products, the study highlights the need for alternative strategies to address these deficiencies.

Keywords: pre-packaged beverages, nutrients content, nutrients declaration, daily percentage value, mandatory addition of vitamins

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Authors: Strahinja Kovacevic, Milica Karadzic Banjac, Jasmina Vitas, Stefan Vukmanovic, Radomir Malbasa, Lidija Jevric, Sanja Podunavac-Kuzmanovic

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The present study deals with chemometric regression analysis of quality parameters and the radical scavenging ability of kombucha fermented kefir-like products obtained with winter savory (WS), peppermint (P), stinging nettle (SN) and wild thyme tea (WT) kombucha inoculums. Each analyzed sample was described by milk fat content (MF, %), total unsaturated fatty acids content (TUFA, %), monounsaturated fatty acids content (MUFA, %), polyunsaturated fatty acids content (PUFA, %), the ability of free radicals scavenging (RSA Dₚₚₕ, % and RSA.ₒₕ, %) and pH values measured after each hour from the start until the end of fermentation. The aim of the conducted regression analysis was to establish chemometric models which can predict the radical scavenging ability (RSA Dₚₚₕ, % and RSA.ₒₕ, %) of the samples by correlating it with the MF, TUFA, MUFA, PUFA and the pH value at the beginning, in the middle and at the end of fermentation process which lasted between 11 and 17 hours, until pH value of 4.5 was reached. The analysis was carried out applying univariate linear (ULR) and multiple linear regression (MLR) methods on the raw data and the data standardized by the min-max normalization method. The obtained models were characterized by very limited prediction power (poor cross-validation parameters) and weak statistical characteristics. Based on the conducted analysis it can be concluded that the resulting radical scavenging ability cannot be precisely predicted only on the basis of MF, TUFA, MUFA, PUFA content, and pH values, however, other quality parameters should be considered and included in the further modeling. This study is based upon work from project: Kombucha beverages production using alternative substrates from the territory of the Autonomous Province of Vojvodina, 142-451-2400/2019-03, supported by Provincial Secretariat for Higher Education and Scientific Research of AP Vojvodina.

Keywords: chemometrics, regression analysis, kombucha, quality control

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Authors: Shirin jalili, Hadi Shirzad, Samaneh Nabavi, Somayeh Khanjani

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Biosensors in forensic analysis are ideal biological tools that can be used for rapid and sensitive initial screening and testing to detect of suspicious components like biological and chemical agent in crime scenes. The wide use of different biomolecules such as proteins, nucleic acids, microorganisms, antibodies and enzymes makes it possible. These biosensors have great advantages such as rapidity, little sample manipulation and high sensitivity, also Because of their stability, specificity and low cost they have become a very important tool to Forensic analysis and detection of crime. In crime scenes different substances such as rape samples, Semen, saliva fingerprints and blood samples, act as a detecting elements for biosensors. On the other hand, successful fluid recovery via biosensor has the propensity to yield a highly valuable source of genetic material, which is important in finding the suspect. Although current biological fluid testing techniques are impaired for identification of body fluids. But these methods have disadvantages. For example if they are to be used simultaneously, Often give false positive result. These limitations can negatively result the output of a case through missed or misinterpreted evidence. The use of biosensor enable criminal researchers the highly sensitive and non-destructive detection of biological fluid through interaction with several fluid-endogenous and other biological and chemical contamination at the crime scene. For this reason, using of the biosensors for detecting the biological fluid found at the crime scenes which play an important role in identifying the suspect and solving the criminal.

Keywords: biosensors, forensic analysis, biological fluid, crime detection

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Authors: Lucia Salvioni, Pietro Giorgio Lovaglio, Valerio Leoni, Miriam Colombo, Luisa Fiandra

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The involvement of plasmatic surfactant protein-D (SP-D) in pulmonary diseases has been long investigated, and over the last two years, more interest has been directed to determine its role as a marker of COVID-19. In this direction, several studies aimed to correlate pulmonary surfactant proteins with the clinical manifestations of the virus indicated SP-D as a prognostic biomarker of COVID-19 pneumonia severity. The present work has performed a retrospective study on a relatively large cohort of patients of Hospital Pio XI of Desio (Lombardia, Italy) with the aim to assess differences in the hematic SP-D concentrations among COVID-19 patients and healthy donors and the role of SP-D as a prognostic marker of severity and/or of mortality risk. The obtained results showed a significant difference in the mean of log SP-D levels between COVID-19 patients and healthy donors, so as between dead and survived patients. SP-D values were significantly higher for both hospitalized COVID-19 and dead patients, with threshold values of 150 and 250 ng/mL, respectively. SP-D levels at admission and increasing differences among follow-up and admission values resulted in the strongest significant risk factors of mortality. Therefore, this study demonstrated the role of SP-D as a predictive marker of SARS-CoV-2 infection and its outcome. A significant correlation of SP-D with patient mortality indicated that it is also a prognostic factor in terms of mortality, and its early detection should be considered to design adequate preventive treatments for COVID-19 patients.

Keywords: SARS-CoV-2 infection, COVID-19, surfactant protein-D (SP-D), mortality, biomarker

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Authors: Daria Grechishnikova, Maria Poptsova

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Transposable elements play an important role in the evolution of various species from bacteria to human. Long Interspersed Nuclear Elements (LINEs) and Short Interspersed Nuclear Elements (SINEs) are two major classes of retrotransposons that occupy a considerable part of any genome and their copy numbers can range form several hundreds to a million. Both LINEs and SINEs multiply through a copy-and-paste mechanism. LINEs encode proteins, which make them capable of self-propagation while SINEs are parasitic and require the machinery of LINEs to multiply. The mechanisms how LINE and SINE RNA is recognized by the LINE-encoded reverse transcriptase (RT) remain unclear. For some SINE-LINE pairs, it was shown that they share a common 3’-end with a stem-loop structure. Majority of the SINE-LINE pairs do not have a common 3’-end. Recently we have shown that in the human genome Alu-L1 pairs have structurally similar stem-loop structure at the 3’-end. Here we extended our analysis to a wide range of species and analyzed LINEs from 161 different species from Repbase and 217 SINE sequences from SINEBase. It appeared that all of the analyzed sequences contained stem-loop structures at the 3’-end. Here we conclude that it is very likely that a common evolutionary mechanism of transposon RNA recognition requires the presence of stem-loop structures at their 3’-end.

Keywords: LINE, SINE, mechanisms of retrotransposition, retrotransposons, stem-loop, stem-loop structures, transposons

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Authors: Saqib Ali, Man-Qun Wang

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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.

Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR

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Authors: Elena K. Schneider, Johnny X. Huang, Vincenzo Carbone, Mark Baker, Mohammad A. K. Azad, Matthew A. Cooper, Jian Li, Tony Velkov

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Ivacaftor is a novel CF trans-membrane conductance regulator (CFTR) potentiator that improves the pulmonary function for cystic fibrosis patients bearing a G551D CFTR-protein mutation. Because ivacaftor is highly bound (>97%) to plasma proteins, there is the strong possibility that co-administered CF drugs that compete for the same plasma protein binding sites and impact the free drug concentration. This in turn could lead to drastic changes in the in vivo efficacy of ivacaftor and therapeutic outcomes. This study compares the binding affinity of ivacaftor and co-administered CF drugs for human serum albumin (HSA) and α1-acid glycoprotein (AGP) using surface plasmon resonance and fluorimetric binding assays that measure the displacement of site selective probes. Due to their high plasma protein binding affinities, drug-drug interactions between ivacaftor are to be expected with ducosate, montelukast, ibuprofen, dicloxacillin, omeprazole and loratadine. The significance of these drug-drug interactions is interpreted in terms of the pharmacodynamic/pharmacokinetic parameters and molecular docking simulations. The translational outcomes of the data are presented as recommendations for a staggered treatment regimen for future clinical trials which aims to maximize the effective free drug concentration and clinical efficacy of ivacaftor.

Keywords: human α-1-acid glycoprotein, binding affinity, human serum albumin, ivacaftor, cystic fibrosis

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Authors: Lu Yang, Keng Po Lai

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Metformin, a well-known clinical drug against diabetes, is found with potential anti-diabetic and anti-obese benefits, as reported in increasing evidences. However, the current clinical and experimental investigations are not to reveal the detailed mechanisms of metformin-anti-obesity/hypertension. We have used the bioinformatics strategy, including network pharmacology and molecular docking methodology, to uncover the key targets and pathways of bioactive compounds against clinical disorders, such as cancers, coronavirus disease. Thus, in this report, the in-silico approach was utilized to identify the hug targets, pharmacological function, and mechanism of metformin against obesity and hypertension. The networking analysis identified 154 differentially expressed genes of obesity and hypertension, 21 interaction genes, and 6 hug genes of metformin treating hypertensive obesity. As a result, the molecular docking findings indicated the potent binding capability of metformin with the key proteins, including interleukin 6 (IL-6) and chemokine (C-C motif) Ligand 2 (CCL2), in hypertensive obesity. The metformin-exerted anti-hypertensive obesity action involved in metabolic regulation, inflammatory reaction. And the anti-hypertensive obesity mechanisms of metformin were revealed, including regulation of inflammatory and immunological signaling pathways for metabolic homeostasis in tissue and microenvironmental melioration in blood pressure. In conclusion, our identified findings with bioinformatics analysis have demonstrated the detailed hug and pharmacological targets, biological functions, and signaling pathways of metformin treating hypertensive obesity.

Keywords: metformin, obesity, hypertension, bioinformatics findings

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Authors: Vivianne Aggestam

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Purpose: Organic dairy products have steadily increased with consumer popularity in recent years in Sweden, permitting more transport activities. The main aim of this study was to compare the transport costs and the environmental emissions made by the organic and conventional dairy production in Sweden. The objective was to evaluate differences and environmental impacts of transport between the two different production systems, allowing a more transparent understanding of the real impact of transport within the supply chain. Methods: A partial attributional Life Cycle Assessment has been conducted based on a comprehensive survey of Swedish farmers, dairies and consumers regarding their transport needs and costs. Interviews addressed the farmers and dairies. Consumers were targeted through an online survey. Results: Higher transport inputs from conventional dairy transportation are mainly via feed and soil management on farm level. The regional organic milk brand illustrate less initial transport burdens on farm level, however, after leaving the farm, it had equal or higher transportation requirements. This was mainly due to the location of the dairy farm and shorter product expiry dates, which requires more frequent retail deliveries. Organic consumers tend to use public transport more than private vehicles. Consumers using private vehicles for shopping trips primarily bought conventional products for which price was the main deciding factor. Conclusions: Organic dairy products that emphasise its regional attributes do not ensure less transportation and may therefore not be a more “climate smart” option for the consumer. This suggests that the idea of localism needs to be analysed from a more systemic perspective. Fuel and regional feed efficiency can be further implemented, mainly via fuel type and the types of vehicles used for transport.

Keywords: supply chains, distribution, transportation, organic food productions, conventional food production, agricultural fossil fuel use

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Authors: Rabaya Basori, Arup K. Raychaudhuri

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In recent years, a variety of semiconductor nanowires (NWs) has been synthesized and used as basic building blocks for the development of electronic and optoelectronic nanodevices. Dielectrophoresis (DEP) has been widely investigated as a scalable technique to trap and manipulate polarizable objects. This includes biological cells, nanoparticles, DNA molecules, organic or inorganic NWs and proteins using electric field gradients. In this article, we have used DEP force to localize nanowire growth by physical vapor deposition (PVD) method as well as control of NW diameter on field assisted growth of the NWs of CuTCNQ (Cu-tetracyanoquinodimethane); a metal-organic charge transfer complex material which is well known of resistive switching. We report a versatile analysis platform, based on a set of nanogap electrodes, for the controlled growth of nanowire. Non-uniform electric field and dielectrophoretic force is created in between two metal electrodes, patterned by electron beam lithography process. Suspended CuTCNQ nanowires have been grown laterally between two electrodes in the vicinity of electric field and dielectric force by applying external bias. Growth and diameter dependence of the nanowires on external bias has been investigated in the framework of these two forces by COMSOL Multiphysics simulation. This report will help successful in-situ nanodevice fabrication with constrained number of NW and diameter without any post treatment.

Keywords: nanowire, dielectrophoretic force, confined growth, controlled diameter, comsol multiphysics simulation

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Authors: Smriti Pahwa, Karishma Vats, Aditi Macwan, Jija Dutt, Sumukhi Vaid

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Age approporiate complementary feeding has been stressed upon for sound young child nutrition and appropriate growth. National Infant and Young Child Feeding guidelines, policies and programs indicate cognizance of the issue taken by the country’s government, policy makers and technical experts. However, it is important that ordinary people, the caregivers of young children too understand the importance of appropriate feeding. For this, an interface might be required where ordinary people could participate in assessing the gaps in IYCF as a first step to take subsequent action. In this context an attempt was made to extrapolate a citizen led learning level survey that has been involving around 25000 ordinary citizens to reach out to 600,000 children annually for over a decade in India. Based on this philosophy of involving ordinary people in simple assessments to produce understandable actionable evidence, a rapid diet assessment tool was developed and collected from caregivers of 90 < 3year children from two urban clusters in Ahmedabad and Baroda, Gujarat. Target sample for pilot was selected after cluster census. Around half the mothers reported that they had not yet introduced water or other fluids to their < 6 month babies. However, about a third were already feeding them food other than mother’s milk. Although complementary feeding was initiated in almost all (95%) children more than 6 months old, frequency was suboptimal in 60%; in 80% cases no measure was taken to either improve energy or nutrient density; only 33% were fed protective foods; Green Leafy Vegetables consumption was negligible (1.4%). Anganwadi food was not consumed. By engaging ordinary people to generate evidence and understand the gaps, such assessments have the potential to be used to generate useful evidence for action at scale as well as locally.

Keywords: citizen led, grass root engagement, IYCF (Infant and Young Child Feeding), rapid diet assessment, under nutrition

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Authors: Endang Purwati Rahayuningsih

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The aim of this study is Enteropathogenic Eschericia coli O157 (EPEC) is one of the pathogen that can cause inflamation and damage intestinal mucosa, which is leading diarrhea. Inflamation in the intestinal mucosa proved by the presence of secretory Immunoglobulin A (sIgA) on the feces. Isolate dadih is Pediococcus pentosaceus (P. pentosaceus) as a probiotic lactic acid bacteria (LAB) is very usefull to improve sIgA and intestinal mucosa. The objective, to determine the dose and duration administration of P. pentosaceus for bowel frequence, sIgA level and height of illeum villi in mice EPEC-induced diarrhea. Method, using Complete Randomized design studies in mice EPEC-induced diarrhea. Mice was classified into 2 factors. A factor (dose of probiotic) and B factor (duration of probiotic observation) consisted of 0 hour, 12 hours, 24 hours and 36 hours. A factor consisted of negative control, positive control (mice induced by EPEC) and 3 different dose experimental mice. The results were a very significant interaction between dose and duration administration of P. pentosaceus. Mean of the most frequent defecation of mice EPEC-induced was 55 graetly reduced into 12 ,after 24 hours administration P. pentosaceus dose 2 x 1010 cfu/g, Mean of sIgA level of mice induced EPEC was 1,60 μg/ml, very significant different (p<0,01). Mean of sIgA level after 24 administration P. pentosaceus dose 2 x 1010cfu/g was 2,65 μg/ml. Mean of height of illeum villi after induced EPEC 53,04 μm with very significant different after 24 hours administration P. pentosaceus dose 2 x 1010cfu/g (142,881μm). This study concluded that P. pentosaceus dose 2 x 1010cfu/g after 24 hours is very beneficial to reduced bowel frequence, increase sIgA level and improve the height illeum villi of mice EPEC-induced diarrhea.

Keywords: Pediococcus pentosaceus, sIgA, enteropathogenic Eschericia coli O157, diarrhea, illeum villi

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Authors: Anuradha Gupta, Neelam Bansal, Varinder Uppal

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The present investigation was made on 35 Indian buffalo fetuses ranging from 0.9 cm to 104 cm curved crown rump length (CVRL). The gross anatomical study revealed that all structures were developed at 13 cm CVRL (87 days) in group I. At 0.9 cm CVRL (32 days) the heart was unseptated and tubular and was clearly divided into common atrial chamber dorsally and primitive ventricle in 1.2 cm CVRL fetus (34 days). Septum primum appeared at 1.9 cm CVRL (37 days), truncal ridges at 2.5 cm CVRL (39 days) and foramen ovale in 3.0 cm CVRL (42 days) buffalo foetuses. At 7.6 cm CVRL (62 days) endocardial cushions fused to form left and right atrioventricular openings and four chambered heart was formed in 8.7 cm CVRL (66 days). Endocardium and epicardium was thicker in atria as compared to ventricles in all the age groups. Myocardium of atria was thin as compared to ventricles in all the age groups and was loosely arranged. Immature hyaline cartilage was first appeared at base of aorta in 62 cm CVRL (213 days) fetuses. Intercalated discs were seen in group III and aorta, pulmonary artery, coronary artery were well appreciated in 3.2 cm CVRL (43 days). Neutral and acid mucopolysaccharides were comparatively more in atria than ventricles. Basic proteins showed strong reaction in atrium and ventricle, and intense in conduction system. Lipids and phospholipids were more in myocardium and conduction system than endocardium and epicardium. All the histochemical moieties were comparatively more in tunica intima than media and adventitia of all the great vessels of heart.

Keywords: buffalo, fetal development, histochemistry, heart

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Authors: Renata Shitakubo, Hanyu Yangcheng, Jay-lin Jane, Fernanda Peroni Okita, Beatriz Cordenunsi

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In the degradation transitory starch model, the enzymatic activity of glucan/water dikinase (GWD) and phosphoglucan/water dikinase (PWD) are essential for the granule degradation. GWD and PWD phosphorylate glucose molecules in the positions C6 and C3, respectively, in the amylopectin chains. This action is essential to allow that β-amylase degrade starch granules without previous action of α-amylase. During banana starch degradation, as part of banana ripening, both α- and β-amylases activities and proteins were already detected and, it is also known that there is a GWD and PWD protein bounded to the starch granule. Therefore, the aim of this study was to quantify both Gluc-6P and Gluc-3P in order to estimate the importance of the GWD-PWD-β-amylase pathway in banana starch degradation. Starch granules were isolated as described by Peroni-Okita et al (Carbohydrate Polymers, 81:291-299, 2010), from banana fruit at different stages of ripening, green (20.7%), intermediate (18.2%) and ripe (6.2%). Total phosphorus content was determinate following the Smith and Caruso method (1964). Gluc-6P and Gluc-3P quantifications were performed as described by Lim et al (Cereal Chemistry, 71(5):488-493, 1994). Total phosphorous content in green banana starch is found as 0.009%, intermediary banana starch 0.006% and ripe banana starch 0.004%, both by the colorimetric method and 31P-NMR. The NMR analysis showed the phosphorus content in C6 and C3. The results by NMR indicate that the amylopectin is phosphorylate by GWD and PWD before the bananas become ripen. Since both the total content of phosphorus and phosphorylated glucose molecules at positions C3 and C6 decrease with the starch degradation, it can be concluded that this phosphorylation occurs only in the surface of the starch granule and before the fruit be harvested.

Keywords: starch, GWD, PWD, 31P-NMR

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Authors: Jarel Elgin Tolentino, Arby Denise Nera, Mary Rose Roco, Angela Vianca Aspa, Nikko Beltran, Else Dapat

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Drug resistance by cells has been the problem in the medical field for decades now. The use of medicinal plants as a source of creating powerful drugs has been nowadays recognized worldwide to treat such resistant diseases. In the present study, the potential for Diospyros philippinensis A. DC. to inhibit growth of both bacteria and cancer cell line was conducted. The leaf crude extracts were screened for the presence of phytochemicals and examined for potential bioactivities by employing several assays like Kirby-Bauer disc diffusion method, DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay for the antibacterial, antioxidant and cytotoxic activities of the extract, respectively. Phytochemical test results of the extracts revealed the presence of alkaloids, flavonoids, saponins, phenols, quinones, cardiac glycosides, phlobatannins, carbohydrate, cardenolides and proteins. The leaf extracts were found to exhibit antibacterial activity against gram-positive bacteria, high antioxidant activity (99.22% ± 0.005) but did not show any sign of cytotoxicity towards HCT116 (ATCC CCL-247). The study therefore concludes that D. philippinensis A. DC. leaf extract can be a source of antibacterial and chemopreventive agents. This claim may be used as basis for future investigation.

Keywords: bioassay, medicinal plants, plant crude extracts, phytochemical screening

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Authors: Michael Graz, Natasha Hurril, Andrew Shearer

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Research on feed supplementation with natural compounds is currently being intensively pursued with a view to improving energy utilisation in ruminants and mitigating the production of methane by these animals. Towards this end, a novel combination of extracts from garlic and bitter orange was therefore selected for trials on the basis of their previously published in vitro anti-methanogenic potential. Three separate in vitro experiments were conducted to determine energy utilisation and greenhouse gas production. These included use of rumen fluid from fistulated cows and sheep in batch culture, the Hohenheim gas test, and the Rusitec technique. Experimental and control arms were utilised, with 5g extracts per kilogram of total dietary dry matter (0.05g/kg active compounds) being used to supplement or not supplement the in vitro systems. Respiratory measurements were conducted on experimental day 1 for the batch culture and Hohenheim gas test and on day 14-21 for the Rusitec Technique (in a 21-day trial). Measurements included methane (CH4) production, total volatile fatty acid (VFA) concentration, molar proportions of acetate, propionate and butyrate and degradation of organic matter (Rusitec). CH4 production was reduced by 82% (±16%), 68% (±11%) and 37% (±4%) in the batch culture, Hohenheim gas test and Rusitec, respectively. Total VFA production was reduced by 13% (±2%) and 2% (±0.1%) in the batch culture and Hohenheim gas test whilst it was increased by 8% (±2%) in the Rusitec. Total VFA production was reduced in all tests between 2 and 10%, whilst acetate production was reduced between 10% and 29%. Propionate production which is an indicator of weight gain was increased in all cases between 16% and 30%. Butyrate production which is considered an indicator of potential milk yield was increased by between 6 and 11%. Degradation of organic matter in the Rusitec experiments was improved by 10% (±0.1%). In conclusion, the study demonstrated the potential of the combination of garlic and citrus extracts to improve digestion, enhance body energy retention and limit CH4 formation in relation to feed intake.

Keywords: citrus, garlic, methane, ruminants

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Authors: Jiun-Nan Hou, Tien-Huang Chen, Wei-June Chen

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Dengue viruses (DENVs) are naturally transmitted between humans by mosquito vectors. Mosquito cells usually survive DENV infection, allowing infected mosquitoes to retain an active status for virus transmission. In this study, we found that DENV2 virus infection in mosquito cells causes the unfolded protein response (UPR) that activates the protein kinase RNA-like endoplasmic reticulum kinase (PERK) signal pathway, leading to shutdown of global protein translation in infected cells which was apparently regulated by the PERK signal pathway. According to observation in this study, the PERK signal pathway in DENV2-infected C6/36 cells alleviates ER stress, and reduces initiator and effector caspases, as well as the apoptosis rate via shutdown of cellular proteins. In fact, phosphorylation of eukaryotic initiation factor 2ɑ (eIF2ɑ) by the PERK signal pathway may impair recruitment of ribosomes that bind to the mRNA 5’-cap structure, resulting in an inhibitory effect on canonical cap-dependent cellular protein translation. The resultant pro-survival “byproduct” of infected mosquito cells is undoubtedly advantageous for viral replication. This finding provides insights into elucidating the PERK-mediated modulating web that is actively involved in dynamic protein synthesis, cell survival, and viral replication in mosquito cells.

Keywords: cap-dependent protein translation, dengue virus, endoplasmic reticulum stress, mosquito cells, PERK signal pathway

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Authors: A. Aslihan Gokaltun, Ali Tuncel

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The unique properties of capillary electrochromatography (CEC) such as high performance, high selectivity, low consumption of both reagents and analytes ensure this technique an attractive one for the separation of biomolecules including nucleosides and nucleotides, peptides, proteins, carbohydrates. Monoliths have become a well-established separation media for CEC in the format that can be compared to a single large 'particle' that does not include interparticular voids. Convective flow through the pores of monolith significantly accelerates the rate of mass transfer and enables a substantial increase in the speed of the separation. In this work, we propose a new approach for the preparation of cationic monolithic stationary phase for capillary electrochromatography. Instead of utilizing a charge bearing monomer during polymerization, the desired charge-bearing group is generated on the capillary monolith after polymerization by using the reactive moiety of the monolithic support via one-pot, simple reaction. Optimized monolithic column compensates the disadvantages of frequently used reversed phases, which are difficult for separation of polar solutes. Rapid separation and high column efficiencies are achieved for the separation of neutral analytes, nucleic acid bases and nucleosides in reversed phase mode. Capillary monolith showed satisfactory hydrodynamic permeability and mechanical stability with relative standard deviation (RSD) values below 2 %. A new promising, reactive support that has a 'ligand selection flexibility' due to its reactive functionality represent a new family of separation media for CEC.

Keywords: biomolecules, capillary electrochromatography, cationic monolith, neutral analytes

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Authors: Sarita Puri, Tapan K. Chaudhuri

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Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.

Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability

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Authors: Mayra Pilamunga, Edwin Vera

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The consumption of dark chocolate is beneficial due to its high content of flavonoids, catechins, and procyanidins. To improve its properties, fortification of chocolate with polyphenols, anthocyanins, soy milk powder and other compounds has been evaluated in several studies. However, to our best knowledge, the addition of carotenes to chocolate has not been tested. Carotenoids, especially ß-carotene and lutein, are widely distributed in fruits and vegetables so that they could be extracted from agro-industrial waste, such as fruit processing. On the other hand, limonene produces crystalline changes of cocoa butter and improves its consistency and viscosity. This study aimed to evaluate the production of dark chocolate with the addition of carotenes extracted from an agro industrial waste and to improve its rheological properties and crystallization, with orange essential oil. The dried and fermented cocoa beans were purchased in Puerto Quito, Ecuador, and had a fat content of 51%. Six types of chocolates were formulated, and two formulations were chosen, one at 65% cocoa and other at 70% cocoa, both with a solid: fat ratio of 1.4:1. With the formulations selected, the influence of the addition of 0.75% and 1.5% orange essential oil was evaluated, and analysis to measure the viscosity, crystallization and sensory analysis were done. It was found that essential oil does not generate significant changes in the properties of chocolate, but has an important effect on aroma and coloration, which changed from auburn to brown. The best scores on sensory analysis were obtained for the samples formulated with 0.75% essential oil. Prior to the formulation with carotenes, the extraction of these compounds from pineapple peels were performed. The process was done with and without a previous enzymatic treatment, with three solid-solvent ratios. The best treatment was using enzymes in a solids-solvent ratio of 1:12.5; the extract obtained under these conditions had 4.503 ± 0.214 μg Eq. β-carotene/mL. This extract was encapsulated with gum arabic and maltodextrin, and the solution was dried using a freeze dryer. The encapsulated carotenes were added to the chocolate in an amount of 1.7% however 60,8 % of them were lost in the final product.

Keywords: cocoa, fat crystallization, limonene, carotenoids, pineapple peels

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Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.

Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon

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