Search results for: whey protein isolated

Authors: Isabella Jabbour

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The world's population is expected to reach over 10 billion by 2050, creating a significant demand for food production, particularly in the agricultural industry. Cellular agriculture presents a solution to this challenge by producing meat that resembles traditionally produced meat, but with significantly less land use. Decellularized plant scaffolds, such as spinach leaves, have been shown to be a suitable edible scaffold for growing animal muscle, enabling cultured cells to grow and organize into three-dimensional structures that mimic the texture and flavor of conventionally produced meat. However, the use of freshwater to remove the intact extracellular material from these plants remains a concern, particularly when considering scaling up the production process. In this study, two protocols were used, 1X SDS and Boom Sauce, to decellularize spinach leaves with both distilled water and ocean water. The decellularization process was confirmed by histology, which showed an absence of cell nuclei, DNA and protein quantification. Results showed that spinach decellularized with ocean water contained 9.9 ± 1.4 ng DNA/mg tissue, which is comparable to the 9.2 ± 1.1 ng DNA/mg tissue obtained with DI water. These findings suggest that decellularized spinach leaves using ocean water hold promise as an eco-friendly and cost-effective scaffold for laboratory-grown meat production, which could ultimately transform the meat industry by providing a sustainable alternative to traditional animal farming practices while reducing freshwater use.

Keywords: cellular agriculture, plant scaffold, decellularization, ocean water usage

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Authors: Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Cheng Chi, Saekil Yun, Sang Guen Kim, Sang Wha Kim, Jeong Woo Kang, Se Jin Han, Se Chang Park

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Vibrio corallilyticus is a well-known pathogen of coral. It is also infectious to a variety of shellfish species, including Pacific oyster (Crassostrea gigas) larvae. V. corallilyticus is remained to be a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facility. Owing to the high mortality and contagious nature of the pathogen, large amount of antibiotics has been used for disease prevention and control. However, indiscriminate use of antibiotics may result in food and environmental pollution, and development of antibiotic resistant strains. Therefore, eco-friendly disease preventative measures are imperative for sustainable bivalve culture. The present investigation proposes the application of bacteriophage (phage) as an effective alternative method for controlling V. corallilyticus infection in marine bivalve hatcheries. Isolation of phages from sea water sample was carried out using drop or double layer agar methods. The host range, stability and morphology of the phage isolates were studied. In vivo phage efficacy to prevent V. corallilyticus infection in oyster larvae was also performed. The isolated phages, named pVco-5 and pVco-7 was classified as a podoviridae and pVco-14, was classified as a siphoviridae. Each phages were infective to four strains of seven V. corallilyticus strains tested. When oyster larvae were pre-treated with the phage before bacterial challenge, mortality of the treated oyster larvae was lower than that in the untreated control. This result suggests that each phages have the potential to be used as therapeutic agent for controlling V. corallilyticus infection in marine bivalve hatchery.

Keywords: bacteriophage, Vibrio coralliilyticus, Oyster larvae, mortality

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Authors: Kalpana Poudel-Tandukar

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Background: Human Immunodeficiency Virus (HIV) infection has been frequently associated with vitamin D deficiency and depression. Vitamin D deficiency increases the risk of depression in people without HIV. We assessed the cross-sectional and prospective associations between serum concentrations of 25-hydroxyvitamin D (25[OH]D) and depression in a HIV-positive people. Methods: A survey was conducted among 316 HIV-positive people aged 20-60 years residing in Kathmandu, Nepal for a cross-sectional association at baseline, and among 184 participants without depressive symptoms at baseline who responded to both baseline (2010) and follow-up (2011) surveys for prospective association. The competitive protein-binding assay was used to measure 25(OH)D levels and the Beck Depression Inventory-Ia method was used to measure depression, with cut off score 20 or higher. Relationships were assessed using multiple logistic regression analysis with adjustment of potential confounders. Results: The proportion of participants with 25(OH)D level of <20ng/mL, 20-30ng/mL, and >30ng/mL were 83.2%, 15.5%, and 1.3%, respectively. Only four participants with 25(OH)D level of >30ng/mL were excluded in the further analysis. The mean 25(OH)D level in men and women were 15.0ng/mL and 14.4ng/mL, respectively. Twenty six percent of participants (men:23%; women:29%) were depressed. Participants with 25(OH)D level of < 20 ng/mL had a 1.4 fold higher odds of depression in a cross-sectional and 1.3 fold higher odds of depression after 18 months of baseline compared to those with 25(OH)D level of 20-30ng/mL (p=0.40 and p=0.78, respectively). Conclusion: Vitamin D may not have significant impact against depression among HIV-positive people with 25(OH)D level below normal ( > 30ng/mL).

Keywords: depression, HIV, Nepal, vitamin D

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Authors: Sara Arafeh, Kovriguine Evguine

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Cytochromes P450 are enzymes that require a supply of electrons to catalyze the synthesis of steroid hormones, fatty acids, and prostaglandin hormone. Cytochrome P450 Oxidoreductase (CYPOR), a membrane bound enzyme, provides these electrons in its open conformation. CYPOR has two cytosolic domains (FAD domain and FMN domain) and an N-terminal in the membrane. In its open conformation, electrons flow from NADPH, FAD, and finally to FMN where cytochrome P450 picks up these electrons. In the closed conformation, cytochrome P450 does not bind to the FMN domain to take the electrons. It was found that when the cytosolic domains are isolated, CYPOR could not bind to cytochrome P450. This suggested that the membrane environment is important for CYPOR function. This project takes the initiative to better understand the dynamics of CYPOR in its full length. Here, we determine the distance between specific sites in the FAD and FMN binding domains in CYPOR by Forster Resonance Energy Transfer (FRET) and Ultrafast TA spectroscopy with and without NADPH. The approach to determine these distances will rely on labeling these sites with red and infrared fluorophores. Mimic membrane attachment is done by inserting CYPOR in lipid nanodiscs. By determining the distances between the donor-acceptor sites in these domains, we can observe the open/closed conformations upon reducing CYPOR in the presence and absence of cytochrome P450. Such study is important to better understand CYPOR mechanism of action in various endosomal membranes including hepatic CYPOR which is vital in plasma cholesterol homeostasis. By investigating the conformational cycles of CYPOR, we can synthesize drugs that would be more efficient in affecting the steroid hormonal levels and metabolism of toxins catalyzed by Cytochrome P450.

Keywords: conformational cycle of CYPOR, cytochrome P450, cytochrome P450 oxidoreductase, FAD domain, FMN domain, FRET, Ultrafast TA Spectroscopy

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Authors: Hafsa Tahir, Sanaullah Iqbal

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Glactooligosaccharide (GOS) is a type of prebiotic which is mainly found in human milk. GOS belongs to those bacteria which stimulates the growth of beneficial bacteria in human intestines. The aim of the present study was to develop a value-added product by producing prebiotic (GOS) in buttermilk through trans galactosylation. Buttermilk is considered as an industrial waste which is discarded after the production of butter and cream. It contains protein, minerals, vitamins and a smaller amount of fat. Raw milk was pasteurized at 100º C for butter production and then trans galactosylation process was induced in the butter milk thus obtained to produce prebiotic GOS. Results showed that the enzyme (which was obtained from bacterial strain of Esecrshia coli and has a gene of Lactobacillus reuteri L103) concentration between 400-600µl/5ml can produce GOS in 30 minutes. Chemical analysis and sensory evaluation of plain and GOS containing buttermilk showed no remarkable difference in their composition. Furthermore, the shelf-life study showed that there was non-significant (P>0.05) difference in glass and pouch packaging of buttermilk. Buttermilk in pouch packaging maintained its stability for 6 days without the addition of preservatives. Therefore it is recommended that GOS enriched buttermilk which is generally considered as a processing waste in dairy manufacturing can be turned into a cost-effective nutritional functional food product. This will not only enhance the production efficiency of butter processing but also will create a new market opportunity for dairy manufacturers all over the world.

Keywords: buttermilk, galactooligosaccharide, shelf Life, transgalactosylation

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Authors: Strahinja Kovačević, Sanja Podunavac-Kuzmanović, Lidija Jevrić, Milica Karadžić

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The present study is based on the quantitative structure-activity relationship (QSAR) analysis of eighteen compounds with anti-prion activity. The structures and anti-prion activities (expressed in response units, RU%) of the analyzed compounds are taken from CHEMBL database. In the first step of analysis 85 molecular descriptors were calculated and based on them the hierarchical cluster analysis (HCA) and principal component analysis (PCA) were carried out in order to detect potential significant similarities or dissimilarities among the studied compounds. The calculated molecular descriptors were physicochemical, lipophilicity and ADMET (absorption, distribution, metabolism, excretion and toxicity) descriptors. The first stage of the QSAR analysis was simple linear regression modeling. It resulted in one acceptable model that correlates Henry's law constant with RU% units. The obtained 2D-QSAR model was validated by cross-validation as an internal validation method. The validation procedure confirmed the model’s quality and therefore it can be used for prediction of anti-prion activity. The next stage of the analysis of anti-prion activity will include 3D-QSAR and molecular docking approaches in order to select the most promising compounds in treatment of prion diseases. These results are the part of the project No. 114-451-268/2016-02 financially supported by the Provincial Secretariat for Science and Technological Development of AP Vojvodina.

Keywords: anti-prion activity, chemometrics, molecular modeling, QSAR

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Authors: Yousef M. Al-Roomi, Kaneez Fatema Hussain

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This work evaluates the thermal degradation kinetic parameters of CaSO₄-complex isolated after the inhibition effect of maleic-anhydride based polymer (YMR-polymers). Pyrolysis experiments were carried out at four heating rates (5, 10, 15 and 20°C/min). Several analytical model-free methods were used to determine the kinetic parameters, including Friedman, Coats and Redfern, Kissinger, Flynn-Wall-Ozawa and Kissinger-Akahira–Sunose methods. The Criado model fitting method based on real mechanism followed in thermal degradation of the complex has been applied to explain the degradation mechanism of CaSO₄-complex. In addition, a simple dynamic model was proposed over two temperature ranges for successive decomposition of CaSO₄-complex which has a combination of organic and inorganic part (adsorbed polymer + CaSO₄.2H₂O scale). The model developed enabled the assessment of pre-exponential factor (A) and apparent activation-energy (Eₐ) for both stages independently using a mathematical developed expression based on an integral solution. The unique reaction mechanism approach applied in this study showed that (Eₐ₁-160.5 kJ/mole) for organic decomposition (adsorbed polymer stage-I) has been lower than Eₐ₂-388 kJ/mole for the CaSO₄ decomposition (inorganic stage-II). Further adsorbed YMR-antiscalant not only reduced the decomposition temperature of CaSO₄-complex compared to CaSO₄-blank (CaSO₄.2H₂O scales in the absence of YMR-polymer) but also distorted the crystal lattice of the organic complex of CaSO₄ precipitates, destroying their compact and regular crystal structures observed from XRD and SEM studies.

Keywords: CaSO₄-complex, maleic-anhydride polymers, thermal degradation kinetics and mechanism, XRD and SEM studies

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Authors: Martynova Alina, Lyubov Buzoleva

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Streptococcus pneumoniae is the causative agent of pneumonias and meningitids throught all the world. Having high genetic diversity, this microorganism can cause different clinical forms of pneumococcal infections and microbiologically it is really difficult diagnosed by routine methods. Also, epidemiological surveillance requires more developed methods of molecular typing because the recent method of serotyping doesn't allow to distinguish invasive and non-invasive isolates properly. Non-coding genome of bacteria seems to be the interesting source for seeking of highly distinguishable markers to discriminate the subspecies of such a variable bacteria as Streptococcus pneumoniae. Technically, we proposed scheme of discrimination of S.pneumoniae strains with amplification of non-coding region (SP_1932) with the following restriction with 2 types of enzymes of Alu1 and Mn1. Aim: This research aimed to compare different methods of typing and their application for molecular epidemiology purposes. Methods: we analyzed population of 100 strains of S.pneumoniae isolated from different patients by different molecular epidemiology methods such as pulse-field gel electophoresis (PFGE), restriction polymorphism analysis (RFLP) and multilolocus sequence typing (MLST), and all of them were compared with classic typing method as serotyping. The discriminative power was estimated with Simpson Index (SI). Results: We revealed that the most discriminative typing method is RFLP (SI=0,97, there were distinguished 42 genotypes).PFGE was slightly less discriminative (SI=0,95, we identified 35 genotypes). MLST is still the best reference method (SI=1.0). Classic method of serotyping showed quite weak discriminative power (SI=0,93, 24 genotypes). In addition, sensivity of RFLP was 100%, specificity was 97,09%. Conclusion: the most appropriate method for routine epidemiology surveillance is RFLP with non-coding region of Streptococcsu pneumoniae, then PFGE, though in some cases these results should be obligatory confirmed by MLST.

Keywords: molecular epidemiology typing, non-coding genome, Streptococcus pneumoniae, MLST

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Authors: Ibrahim Sani, J. T. Amodu, M. R. Hassan, R. J. Tanko, N. Adamu

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This study investigated the digestibility of Brachiaria ruziziensis and Centrosema pascuorum hay mixtures at varying proportions in Yankasa rams. Twelve Yankasa rams with average initial weight 10.25 ± 0.1 kg were assigned to three dietary treatments of B. ruziziensis and C. pascuorum hay at different mixtures (75BR:25CP, 50BR:50CP and 25BR:75CP, respectively) in a Completely Randomized Design (CRD) for a period of 14 days. Concentrate diet was given to the experimental animals as supplement at fixed proportion, while the forage mixture (basal diet) was fed at 3% body weight. Animals on 50BR:50CP had better nutrient digestibility (crude protein, acid and neutral detergent fibre, ether extract and nitrogen free extract) than other treatment diets, except in dry matter digestibility (87.35%) which compared with 87.54% obtained in 25BR:75CP treatment diet and also organic matter digestibility. All parameters taken on nitrogen balance with the exception of nitrogen retained were significantly higher (P < 0.05) in animals fed 25BR:75CP diet, but were statistically similar with values obtained for animals on 50BR:50CP diet. From results obtained in this study, it is concluded that mixture of 25%BR75%CP gave the best nutrient digestibility and nitrogen balance in Yankasa rams. It is therefore recommended that B. ruziziensis and C. pascuorum should be fed at 50:50 mixture ratio for enhanced animal growth and performance in Nigeria.

Keywords: B. ruziziensis, C. pascuorum, digestibilty, rams, Yankasa

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Authors: Samane Maroufi, Rasoul Khayyam Nekouei, Sajjad Sefimofarah, Veena Sahajwalla

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The present work details a three-stage strategy based on selective purification of rare earth oxide (REOs) isolated from end-of-life nickel-metal hydride (Ni-MH) batteries leading to high-yield fabrication of defect-rich Mn1-x-y(CeₓLaᵧ)O2-δ film. In step one, major impurities (Fe and Al) were removed from a REE-rich solution. In step two, the resulting solution with trace content of Mn was further purified through electrodeposition which resulted in the synthesis of a non-stoichiometric Mn₋₁₋ₓ₋ᵧ(CeₓLaₓᵧ)O2-δ ultra-thin film, with controllable thicknesses (5-650 nm) and transmittance (~29-100%)in which Ce4+/3+ and La3+ ions were dissolved in MnO2-x lattice. Due to percolation impacts on the optoelectronic properties of ultrathin films, a representative Mn1-x-y(CexLay)O2-δ film with 86% transmittance exhibited an outstanding areal capacitance of 3.4 mF•cm-2, mainly attributed to the intercalation/de-intercalation of anionic O2- charge carriers through the atomic tunnels of the stratified Mn1-x-y(CexLay)O2-δ crystallites. Furthermore, the Mn1-x-y(CexLay)O2-δ exhibited excellent capacitance retention of ~90% after 16,000 cycles. Such stability was shown to be associated with intervalence charge transfers occurring among interstitial Ce/La cations and Mn oxidation states within the Mn₋₁₋ₓ₋ᵧ(CexLay)O2-δ structure. The energy and power densities of the transparent flexible Mn₋₁₋ₓ₋ᵧ(CexLay)O2-δ full-cell pseudocapacitor device with a solid-state electrolyte was measured to be 0.088 µWh.cm-2 and 843 µW.cm-2, respectively. These values showed insignificant changes under vigorous twisting and bending to 45-180˚, confirming these materials are intriguing alternatives for size-sensitive energy storage devices. In step three, the remaining solution purified further, that led to the formation of REOs (La, Ce, and Nd) nanospheres with ~40-50 nm diameter.

Keywords: spent Ni-MH batteries, green energy, flexible pseudocapacitor, rare earth elements

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Authors: Constantine Uwaremwe, Wenjie Bao, Bachir Goudia Daoura, Sandhya Mishra, Xianxian Zhang, Lingjie Shen, Shangwen Xia, Xiaodong Yang

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Background: Plukenetia volubilis Linneo is an oleaginous plant belonging to the family Euphorbiaceae. Due to its seeds containing a high content of edible oil and rich in vitamins, P. volubilis is cultivated as an economical plant worldwide. However, the cultivation and growth of P. volubilis is challenged by phytopathogen invasion leading to production loss. Methods: In the current study, we tested the pathogenicity of fungal pathogens isolated from root rot infected P. volubilis plant tissues by inoculating them into healthy P. volubilis seedlings. Metagenomic sequencing was used to assess the shift in the fungal community of P. volubilis rhizosphere soil after root rot infection. Results: Four Fusarium isolates and two Rhizopus isolates were found to be root rot causative agents of P. volubilis as they induced typical root rot symptoms in healthy seedlings. The metagenomic sequencing data showed that root rot infection altered the rhizosphere fungal community. In root rot infected soil, the richness and diversity indices increased or decreased depending on pathogens. The four most abundant phyla across all samples were Ascomycota, Glomeromycota, Basidiomycota, and Mortierellomycota. In infected soil, the relative abundance of each phylum increased or decreased depending on the pathogen and functional taxonomic classification. Conclusions: Based on our results, we concluded that Fusarium and Rhizopus species cause root rot infection of P. volubilis. In root rot infected P. volubilis, the shift in the rhizosphere fungal community was pathogen-dependent. These findings may serve as a key point for a future study on the biocontrol of root rot of P. volubilis.

Keywords: fusarium spp., plukenetia volubilis l., rhizopus spp., rhizosphere fungal community, root rot

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Authors: Sunday Sedodo Nupo, Clara Berstien Oguntona, Babatunde Oguntona, Oluseyi Akinloye, P. A. Olunusi Adeboye

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Cardiovascular diseases are now becoming dominant sources of morbidity and mortality worldwide. This study investigated the lipid profile of civil servants. A cross-sectional study was carried out among randomly selected 202 male and 298 female civil servants in Abeokuta Ogun state. A pretested structured questionnaire was used to elicit information on history of non-communicable diseases and physical activity pattern of the respondents. The blood pressures of the subjects were measured and classified using World Health Organization criteria. The total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL). Ethical approval was obtained from Ogun State Ministry of Health. Data collected were analysed using Statistical package for social science version 17.1. Results showed that majority (76%) of the subjects were within the age range of 20 - 40 years, 75% earned between N58,500 - N98,000 monthly and 68% were sedentary. The mean energy intake of men and women were 3942±38 kcal and 2791±3 kcal respectively, while the protein intake for men was 65±49 g/day and 54.28±40 g/day for women. Desirable TC level (<200 mg/dl) was found in 80% of the selected subjects while the normal TG (<150 mg/dl) and LDL (<129 mg/dl) was found in 95% and 90% subjects respectively. The mean TC was 78.91±11 mg/dl and 62.69±9 mg/dl in men and women respectively. The study showed that most of the subjects had normal lipid in terms of serum triglycerides, total cholesterol, HDL cholesterol and LDL cholesterol.

Keywords: high density lipoprotein, morbidity, mortality, triglycerides

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Authors: Stephen Olusanmi Akintayo, Ilesanmi Fadahunsi

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The ability of Lactic acid bacteria (LAB) to grow and survive in milk is being exploited in industrial and biotechnological applications. Although considerable studies have been reported on the fermentation of milk, however, not so much work has been documented on the selection of LAB strains from milk of the Nigerian local cattle breeds for their starter culture potentials. A total of 110 LAB were isolated from raw milk of Sokoto gudali cattle breed. The isolates were screened for their proteolytic activities on skimmed milk media with isolates A07, F06 and A01 showing the highest zone of clearance of 18.5mm, 18.5mm, and 18.0mm respectively and were selected for the studies of their growth in different constituents of milk. A01, F06, and A07 were identified as Pediococcus acidilactici, Lactococcus raffinolactis, and Leuconostoc mesenteriodes respectively using cultural, biochemical, physiological and molecular characterization techniques. Leuconostoc mesenteriodes showed the highest growth in all the milk components that were used in this study. The three LAB species selected showed a growth range of 6.46 log cfu/ml to 10.91 log cfu/ml in lactose with Leuconostoc mesenteriodes showing the highest growth of 10.91 log cfu/ml while Pediococcus acidilactici recorded the lowest growth of 9.78 log cfu/ml. In medium containing leucine as the only amino acid, the viable counts of Pediococcus acidilactici, Lactococcus raffinolactis and Leuconostoc mesenteriodes in log cfu/ml at zero hour were 6.39, 6.36 and 6.38 respectively which increased to 9.31 log cfu/ml, 9.21 log cfu/ml, 9.92 log cfu/ml respectively after 24 hours. Similarly, in all other substrates (casein, lysine, glutamic acid, aspartic acid, stearic acid and oleic acid ) tested in this study, Leuconostoc mesenteriodes showed the highest growth. It was observed that the highest quantity of lactic acid (15.31mg/ml) was produced by Leuconostoc mesenteriodes. The same trend was also observed in the production of diacetyl and hydrogen peroxide by the three tested microorganisms. Due to its ability to grow maximally in milk components, Leuconostoc mesenteriodes shows potential as starter culture for milk fermentation.

Keywords: Leuconostoc mesenteriodes, lactic acid bacteria, Sokoto gudali, starter culture

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Authors: W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel

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Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutamine-rich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.

Keywords: Candida albicans, morphogenetic development, TEA domain, hyphal formation, TEC1

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Authors: Muhammad Sohail Sajid, Qurat-ul-Ain, Zafar Iqbal, Muhammad Nisar Khan, Asma Kausar, Adil Ejaz

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Rickettsiosis, caused by a Spotted Fever Group Rickettsiae (SFGR), is considered as an emerging infectious disease from public and veterinary perspective. The present study reports distribution of SFGR in the host (buffalo and cattle) and vector (ticks) population determined through gene specific amplification through PCR targeting outer membrane protein (ompA). Tick and blood samples were collected using standard protocols through convenient sampling from district Faisalabad. Ticks were dissected to extract salivary glands (SG). Blood and tick SG pools were subjected to DNA extraction and amplification of ompA using PCR. Overall prevalence of SFGR was reported as 21.5% and 33.6 % from blood and ticks, respectively. Hyalomma anatolicum was more prevalent tick associated with SFGR as compared to Rhipicephalus sp. Higher prevalence of SFGR was reported in cattle (25%) population as compared to that of buffalo (17.07%). On seasonal basis, high SFGR prevalence was recorded during spring season (48.1%, 26.32%, 17.76%) as compared to winter (27.9%, 21.43%, 15.38%) in vector and host (cattle and buffalo respectively) population. Sequencing analysis indicated that rickettsial endo-symbionts were associated with ticks of the study area. These results provided baseline information about the prevalence of SFGR in vector and host population.

Keywords: Rickettsia, livestock, polymerase chain reaction, sequencing, ticks, vectors

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Authors: Elina Brahma Hazarika, Jeuti Basumatary, Deepanka Saikia, Jaydeep Das, Micky Moni D'mary, Fungkha Basumatary

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This study includes development of two varieties of biscuits incorporated with: the seeds of Jack fruit (Artocarpus heterophyllus), which post-consumption of it’s pulp, is discarded as a waste, and Cassava (Manihot esculenta) tubers.The jack fruit seeds and cassava were first ground into flour and its proximate and physiochemical properties were studied. The biscuits that were developed incorporating them had 50% wheat flour and 50% jackfruit seed flour and 50% cassava flours as the major composition, apart from the other general ingredients use in making biscuits. Various trials of compositions were made for baking to get the overall desirable acceptability in biscuits through sensory evaluation. Finally, the best composition of ingredients was selected to make the biscuits, and hence studies were done accordingly to compare it with the properties of their respective raw flours. The results showed that the proximate composition of the biscuits fared better than that of their respective flours: There was a decrease in the Moisture content of both Jackfruit Seed Biscuits and Cassava Biscuits to 4.5% and 6.7% than that of their respective raw flours (8 and 12%). Post-baking, there is increase in the percentages of ash, protein, and fibre contents in both Jackfruit Seed Biscuits and Cassava Biscuits; the values being 3% and 3.8%, 13.2% and 3.3%, and 3.2 and 4.1% respectively. Also the total carbohydrate content in Jackfruit Seed Biscuits and Cassava Biscuits were 66.7% and 71.7% respectively. Their sensory evaluation and texture study also yielded a clear review that they have an overall good acceptability.

Keywords: baking, proximate, sensory, texture

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Authors: Yan-Shiang Chiou, Yi-Huang Hsueh

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Bacillus cereus is a foodborne pathogen that causes two types of foodborne illness, the emetic and diarrheal syndromes. B. cereus consistently ranks among the top three among bacterial foodborne outbreaks in the ten years of 2001 to 2010 in Taiwan. Foodborne outbreak caused by B. cereus has been increased, and recently it ranks second foodborne pathogen after Vibrio parahaemolyticus. This pathogen is difficult to control due to its ubiquitousness in the environment, the psychrotrophic nature of many strains, and the heat resistance of their spores. Because complete elimination of biofilms is difficult, a better understanding of the molecular mechanisms of biofilm formation by B. cereus will help to develop better strategies to control this pathogen. Surface translocation can be an important factor in biofilm formation. In B. cereus, NprR is a quorum sensor, and its apo NprR is a dimer and changes to a tetramer in the presence of NprX. The small peptide NprX may induce conformational change allowing the apo dimer to switch to an active tetramer specifically recognizing target DNA sequences. Our result showed that mutation of nprRX causes surface spreading deficiency. Mutation of flagella, pili and surfactant genes (flgAB, bcpAB, krsABC), did not abolish spreading motility. Under nprRX mutant, mutation of spo0A restored the spreading deficiency. This suggests that spreading motility is not related surfactant, pili and flagella but other unknown mechanism and Spo0A, a sporulation initiation protein, inhibits spreading motility.

Keywords: Bacillus cereus, nprRX, spo0A, spreading motility

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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: Marina Lazar Chandy, N. Kannan, Rajendra Patil, Vinod Mathew, Ajmal Mohamed, P. K. Sreeja, Renju Jose

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Introduction- Arecoline, a major constituent of arecanut has shown to have some effect on liver. The use of arecanut is found to be the most common etiological factor for the development of Oral Submucous fibrosis (O.S.M.F). The effect of arecanut usage on liver in patients with O.S.M.F needs to be assessed. Lipids play a role in structural maintenance of cell. Alterations of lipid profile were noted in cancer patients. O.S.M.F being a precancerous lesion can have some effect on the level of lipids in the body. Objectives: This study was done to assess the alterations in liver enzymes (Serum Glutamic Pyruvic Transaminase(S.G.P.T ,Serum Glutamic Oxaloacetic Transaminase(S.G.O.T)) and lipid metabolism (High Density Lipoprotien(H.D.L) and Apo Lipoprotien A1 (Apo A1)) in patients with O.S.M.F. Methods-130 patients were taken for the study,100 patients with O.S.M.F and 30 as control group without O.S.M.F. Fasting blood sugar levels were taken, centrifuged and analyzed for S.G.P.T,S.G.O.T, H.D.L and Apo A1 using semi automated spectrophotometer. Results: After statistical analysis, it was concluded that there is an elevation of levels of S.G.P.T, S.G.O.T, and decreased levels of H.D.L, Apo A1 for O.S.M.F group when compared with control group. With increased grade of O.S.M.F. and duration of habit, S.G.P.T. & S.G.O.T. increased whereas, H.D.L. & Apo A1 decreased. All the values were statistically significant at p<0.01.

Keywords: apolipoprotien A1, high density lipoprotien, oral submucous fibrosis, serum glutamic oxaloacetic transaminase

Procedia PDF Downloads 327

Authors: Abdelkarim Mahdhi, Fdhila Kais, Faouzi Lamari, Zeineb Hmila, Fathi Kamoun, Maria Ángeles Esteban, Amina Bakhrouf

Abstract:

Aquaculture is the world’s fastest growing food-production sector. However, one of the most serious problems regarding the culture of marine fishes is the mortality associated with pathogenic bacteria that occurs in the critical phases of larval development. Conventional approaches, such as the use of antimicrobial drugs to control diseases, have had limited success in the prevention or cure of aquatic diseases. Promising alternatives to antibiotics are probiotics, which are food supplements consisting of live microorganisms that benefit the host organism. In the search for more effective and environmentally friendly treatments with probionts against pathogenic species in shrimp larval culture, the probiotic properties of Bacillus strains isolated from Artemia culture such as antibacterial activity, adhesion, pathogenicity, toxicity and the effect of marine stress on viability and survival were investigated, as well as the changes occurring in their properties. Analyses showed that these bacteria corresponded to the genus Bacillus sp. Antagonism and adherence assays revealed that these strains have an inhibitory effect against pathogenic bacteria in vitro and in vivo conditions and are fairly adherent. Challenge tests performed with Artemia larvae provided evidence that the tested Bacillus strains were neither pathogenic nor toxic to the host. The tested strains maintained their viability and their probiotic properties during the period of study. The results suggest that the tested strains have suffered changes allowing them to survive in seawater in the absence of nutrients and outside their natural host, identifying them as potential probiotic candidates for Artemia culture.

Keywords: bacillus, probiotic, cell viability, stress response

Procedia PDF Downloads 386

Authors: Andrey V. Khromov, Antonida V. Makhotenko, Ekaterina A. Snigir, Svetlana S. Makarova, Natalia O. Kalinina, Valentin V. Makarov, Mikhail E. Taliansky

Abstract:

Due to its simplicity, CRISPR/Cas9 has become widely used and capable of inducing mutations in the genes of organisms of various kingdoms. The aim of this work was to develop applications for the efficient modification of DNA coding sequences of phytoene desaturase (PDS), coilin and vacuolar invertase (Solanum tuberosum) genes, and to develop a new nanoparticles carrier efficient technology to deliver the CRISPR/Cas9 system for editing the plant genome. For each of the genes - coilin, PDS and vacuolar invertase, five single RNA guide (sgRNAs) were synthesized. To determine the most suitable nanoplatform, two types of NP platforms were used: magnetic NPs (MNPS) and gold NPs (AuNPs). To test the penetration efficiency, they were functionalized with fluorescent agents - BSA * FITS and GFP, as well as labeled Cy3 small-sized RNA. To measure the efficiency, a fluorescence and confocal microscopy were used. It was shown that the best of these options were AuNP - both in the case of proteins and in the case of RNA. The next step was to check the possibility of delivering components of the CRISPR/Cas9 system to plant cells for editing target genes. AuNPs were functionalized with a ribonucleoprotein complex consisting of Cas9 and corresponding to target genes sgRNAs, and they were biolistically bombarded to axillary buds and apical meristems of potato plants. After the treatment by the best NP carrier, potato meristems were grown to adult plants. DNA isolated from this plants was sent to a preliminary fragment of the analysis to screen out the non-transformed samples, and then to the NGS. The present work was carried out with the financial support from the Russian Science Foundation (grant No. 16-16-04019).

Keywords: biobombardment, coilin, CRISPR/Cas9, nanoparticles, NPs, PDS, sgRNA, vacuolar invertase

Procedia PDF Downloads 317

Authors: Amira, A. Goma, U. E. Mahrous

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Welfare may be considered to be a subjective experience; it has a biological function that is related to the fitness and survival of the animal accordingly, researches have suggested that welfare is compromised when the animal's evolutionary fitness is reduced. This study was carried out to explain the effect of some managerial stressors as handling and restraint on behavioral patterns and biochemical parameters of rats. A total of 24 (12 males and 12 females) Sprague-Dawley rats (12 months and 150-180g) were allotted into 3 groups, handled group (4 male and 4 female), restrained group (4 male and 4 female) and control group (4 males and 4 females). The obtained results revealed that time spent feeding, drinking frequency, movement and cage exploration increased significantly in handled rats than other groups, while lying time and licking increased significantly in restrained rats than handled and controls. Moreover, social behavior decreased in both stressed groups than control. Triglycerides were significantly increased in handled rats than other groups, while total lipid, total protein and globulin significantly increased in both treated groups than control. Corticosterone increased in restrained and handled rats than control ones. Moreover, there was an increment in packed cell volume significantly in restrained rats than others. These deducted that if we want to study the effect of stress on animal welfare it is necessary to study the effect of such stressors on animal’s behavior and physiological responses.

Keywords: handling, restraint, welfare, rat, behavior, physiology

Procedia PDF Downloads 452

Authors: Zebiri Saliha

Abstract:

Ochratoxin A (OTA) is a mycotoxin produced by certain species of the genera Aspergillus and Penicillium, primarily found in cereals, coffee, and grapevine products. Its accumulation in the body can lead to nephrotoxic, teratogenic, immunosuppressive, and carcinogenic effects. The objective of this study is to investigate the contamination of consumed wheat and its derivatives by toxic fungi in Algeria. For this purpose, an analysis of 200 samples was conducted, including 90 samples of durum wheat and common wheat and 110 samples of wheat derivatives collected from mills (semolina and flour manufacturers). The results revealed an average fungal contamination rate ranging from 60% to 100%. The identified fungal isolates primarily belonged to the genera Aspergillus (70%), Penicillium (27.5%), Alternaria (40%), and Mucor (19.4%). The density of the fungal flora was higher in products intended for animal consumption, such as durum wheat flour (2525 CFU/g), wheat scraps (3175 CFU/g), and wheat bran (2950 CFU/g). Conversely, low fungal density was observed in fine semolina (900 CFU/g) and flour (800 CFU/g) intended for human consumption. The genus Penicillium was isolated in 46% of the analyzed samples of durum wheat derivatives and in 62.7% of the analyzed samples of common wheat derivatives. The Aspergillus genus dominated the majority of the analyzed samples. Molecular identification of Aspergillus and Penicillium isolates by sequencing ITS1-5.8S-ITS2 regions of DNAr and a part of the calmodulin (CaM) gene indicated that the species involved in the production of OTA in wheat and its derivatives were mainly Aspergillus ochraceus, A. westerdijkia, A. alliaceus, A. carbonarius, and Penicillium islandicus. The amounts of OTA produced by these species were determined by HPLC-FLD and ranged between 0,8.9 and 3033μg/g. Given that food safety and quality are major concerns today, understanding the microbial biodiversity of wheat is crucial because it is a staple food in Algeria.

Keywords: wheat derivatives, Aspergillus, microbial biodiversity, OTA

Procedia PDF Downloads 54

Authors: Emad Hafez, Mahmoud Seleiman

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Two field experiments were carried out in order to investigate the effect of chemical inducers [benzothiadiazole 0.9 mM L-1, oxalic acid 1.0 mM L-1, salicylic acid 0.2 mM L-1] on physiological and technological traits as well as on yields and antioxidant enzyme activities of barley grown under abiotic stress (i.e. water surplus and deficit conditions). Results showed that relative water content, leaf area, chlorophyll and yield as well as technological properties of barley were improved with chemical inducers application under water surplus and water-stress conditions. Antioxidant enzymes activity (i.e. catalase and peroxidase) were significantly increased in barley grown under water-stress and treated with chemical inducers. Yield and related parameters of barley presented also significant decrease under water-stress treatment, while chemical inducers application enhanced the yield-related traits. Starch and protein contents were higher in plants treated with salicylic acid than in untreated plants when water-stress was applied. In conclusion, results show that chemical inducers application have a positive interaction and synergetic influence and should be suggested to improve plant growth, yield and technological properties of water stressed barley. Salicylic acid application was better than oxalic acid and benzothiadiazole in terms of plant growth and yield improvement.

Keywords: antioxidant enzymes, drought stress, Hordeum vulgare L., quality, yield

Procedia PDF Downloads 305

Authors: Mina Kargozari, Isabel Revilla Martin, Ángel A. Carbonell-Barrachina

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Different formulations of Turkish fermented sausages (sucuks) prepared with camel meat-hump (CH), camel meat-beef fat (CB), beef-hump (BH) and beef-beef fat (BB), were characterized. The sausages were analytically compared to determine differences in proximate composition and total cholesterol content (TCC), quality parameters such as fatty acids profile and fat quality characteristics, and lipid oxidation parameters including peroxide value, thiobarbituric acid-reactive substances (TBARS) and resulted carbonyl compounds. The PUFAs/SFAs ratio was higher in CB and BB samples than CH and BH (p<0.05). The higher calculated atherogenic and thrombogenic indexes (AI and TI) were obtained from the samples made with hump (p< 0.05) as a result of high amounts of their SFAs. The CH sausages contained high amount of total fat (p<0.05) among all samples. The CB sucuks exhibited the highest protein content and the lowest TCC and rancidity at the end of ripening (p<0.05). The TBARS results showed that beef fat samples were more susceptible to lipid oxidation. Moreover, no significant difference (p<0.05) was observed for the values of short aldehydes among the sucuk samples excepting nonanal. This study demonstrated that supplementing camel meat for the production of dry-fermented sausage resulted in high quality products with good functional and nutritional characteristics.

Keywords: fermented sausages, quality properties, SPME, total cholesterol content

Procedia PDF Downloads 326

Authors: Naveed Ahmad Fazili

Abstract:

More than 20 human diseases involve the fibrillation of a specific protein/peptide which forms pathological deposits at various sites. Hereditary lysozyme amyloidosis is a systemic disorder which mostly affects liver, spleen and kidney. This conformational disorder is featured by lysozyme fibril formation. In vivo lysozyme fibrillation was simulated under in vitro conditions using a strong denaturant GdHCl at 3M concentration. Sharp decline in the ANS fluorescence intensity compared to the partially unfolded states, almost 20 fold increase in ThT fluorescence intensity, increase in absorbance at 450 nm suggesting turbidity, negative ellipticity peak in the far-UVCD at 217 nm, red shift of 50 nm compared to the native state in congo red assay and appearance of a network of long rope like fibrils in TEM analysis suggested HEWL fibrillation. Anti-fibrillation potency of baicalein against the preformed fibrils of HEWL was investigated following ThT assay in which there was a dose dependent decrease in ThT fluorescence intensity compared to the fibrillar state of HEWL with the maximum effect observed at 150 μM baicalein concentration, loss of negative ellipticity peak in the far-UVCD region, dip in the Rayleigh scattering intensity and absorbance at 350 nm and 450 nm respectively together with a reduction in the density of fibrillar structure in TEM imaging. Thus, it could be suggested that baicalein could prove to be a positive therapeutics for hereditary human lysozyme amyloidosis.

Keywords: amyloid fibrils, baicalein, congo red, negative ellipticity, therapeutics

Procedia PDF Downloads 294

Authors: H. Cano, P. Ferrer, N. Garcia, M. Popovic, J. Zapata

Abstract:

Positive Reinforcement Training is a well-known methodology which has been reported frequently to be used in captive non-human primates. As a matter of fact, it is an invaluable tool for different purposes related with animal welfare, such as primate husbandry and environmental enrichment. It is also essential to perform some cognitive experiments. The main propose of this pilot study was to establish an efficient protocol to train captive olive baboons (Papio anubis). This protocol seems to be vital in the context of a larger research program in which it will be necessary to train a complete population of around 40 baboons. Baboons were studied at the Veterinary Research Farm of the University of Murcia. Temporally isolated animals were trained to perform three basic tasks. Firstly, they were required to take food prices directly from the researchers’ hands. Then a clicker sound or bridge stimulus was added each time the animal acceded to the reinforcement. Finally, they were trained to touch a target, consisted of a whip with a red ball in its end, with their hands or their nose. When the subject completed correctly this task, it was also exposed to the bridge stimulus and awarded with a food price, such as a portion of banana, orange, apple, peach or a raisin. Two protocols were tested during this experiment. In both of them, there were 6 series of 2min training periods each day. However, in the first protocol, the series consisted in 3 trials, whereas in the second one, in each series there were 5 trials. A reliable performance was obtained with only 6 days of training in the case of the 5-trials protocol. However, with the 3-trials one, 26 days of training were needed. As a result, the 5-trials protocol seems to be more effective than the 3-trials one, in order to teach these three basic tasks to olive baboons. In consequence, it will be used to train the rest of the colony.

Keywords: captive primates, olive baboon, positive reinforcement training, Papio anubis, training

Procedia PDF Downloads 126

Authors: Wanwisa Deenin, Abdulhadee Yakoh, Chahya Kreangkaiwal, Orawon Chailapakul, Kanitha Patarakul, Sudkate Chaiyo

Abstract:

LipL32 is an outer membrane protein present only on pathogenic Leptospira species, which are the causative agent of leptospirosis. Leptospirosis symptoms are often misdiagnosed with other febrile illnesses as the clinical manifestations are non-specific. Therefore, an accurate diagnostic tool for leptospirosis is indeed critical for proper and prompt treatment. Typical diagnosis via serological assays is generally performed to assess the antibodies produced against Leptospira. However, their delayed antibody response and complicated procedure are undoubtedly limited the practical utilization especially in primary care setting. Here, we demonstrate for the first time an early-stage detection of LipL32 by an integrated lateral-flow immunoassay with electrochemical readout (eLFIA). A ferrocene trace tag was monitored via differential pulse voltammetry operated on a smartphone-based device, thus allowing for on-field testing. Superior performance in terms of the lowest detectable limit of detection (LOD) of 8.53 pg/mL and broad linear dynamic range (5 orders of magnitude) among other sensors available thus far was established. Additionally, the developed test strip provided a straightforward yet sensitive approach for diagnosis of leptospirosis using the collected human sera from patients, in which the results were comparable to the real-time polymerase chain reaction technique.

Keywords: leptospirosis, electrochemical detection, lateral flow immunosensor, point-of-care testing, early-stage detection

Procedia PDF Downloads 96

Authors: Rachel Shukrun, Szilvia Baron, Victoria Fidel, Anna Shusterman, Osnat Sher, Netanya Kollender, Dror Levin, Yair Peled, Yair Gortzak, Yoav Ben-Shahar, Revital Caspi, Sagi Gordon, Michal Manisterski, Ronit Elhasid

Abstract:

Ewing sarcoma (EWS) is a highly aggressive cancer with a survival rate of 70–80% for patients with localized disease and under 30% for those with metastatic disease. Tumor-infiltrating neutrophils (TIN) can generate extracellular net-like DNA structures known as neutrophil extracellular traps (NETs). However, little is known about the presence and prognostic significance of tumor-infiltrating NETs in EWS. Herein, we investigated 46 patients diagnosed with EWS and treated in the Tel Aviv Medical Center between 2010 and 2021. TINs and NETs were identified in diagnostic biopsies of EWS by immunofluorescent. In addition, NETs were investigated in neutrophils isolated from peripheral blood samples of EWS patients at diagnosis and following neoadjuvant chemotherapy. The relationships between the presence of TINs and NETs, pathological and clinical features, and outcomes were analyzed. Our results demonstrate that TIN and NETs at diagnosis were higher in EWS patients with metastatic disease compared to those with local disease. High NETs formation at diagnosis predicted poor response to neo-adjuvant chemotherapy, relapse, and death from disease (P < .05). NETs formation in peripheral blood samples at diagnosis was significantly elevated among patients with EWS compared to pediatric controls and decreased significantly following neoadjuvant chemotherapy. In conclusion, NETs formation seems to have a role in the EWS immune microenvironment. Their presence can refine risk stratification, predict chemotherapy resistance and survival, and serve as a therapeutic target in patients with EWS.

Keywords: Ewing sarcoma, tumor microenvironment, neutrophil, neutrophil extracellular traps (NETs), prognosis

Procedia PDF Downloads 65

Authors: Athicha Cherdpunt, Patompong Satapornpong

Abstract:

The medical cannabis is made up of components also known as cannabinoids, which consists of two ingredients which are Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD). Interestingly, the Cannabinoid can be used for many treatments such as chemotherapy, including nausea and vomiting, cachexia, anorexia nervosa, spinal cord injury and disease, epilepsy, pain, and many others. However, the adverse drug reactions (ADRs) of THC can cause sedation, anxiety, dizziness, appetite stimulation and impairments in driving and cognitive function. Furthermore, genetic polymorphisms of CYP2C9, CYP2C19 and CYP3A4 influenced the THC metabolism and might be a cause of ADRs. Particularly, CYP2C19*17 allele increases gene transcription and therefore results in ultra-rapid metabolizer phenotype (UM). The aim of this study, is to investigate the frequency of CYP2C19*17 alleles in Thai patients who have been treated with medical cannabis. We prospectively enrolled 60 Thai patients who were treated with medical cannabis and clinical data from College of Pharmacy, Rangsit University. DNA of each patient was isolated from EDTA blood, using the Genomic DNA Mini Kit. CYP2C19*17 genotyping was conducted using the real time-PCR ViiA7 (ABI, Foster City, CA, USA). 30 patients with medical cannabis-induced ADRs group, 20 (67%) were female, and 10 (33%) were male, with an age range of 30-69 years. On the other hand, 30 patients without medical cannabis-induced ADRs (control group) consist of 17 (57%) female and 13 (43%) male. The most ADRs for medical cannabis treatment in the case group were dry mouth and dry throat (77%), tachycardia (70%), nausea (30%) and arrhythmia(10%). Accordingly, the case group carried CYP2C19*1/*1 (normal metabolizer) approximately 93%, while 7% patients carrying CYP2C19*1/*17 (ultra rapid metabolizers) exhibited in this group. Meanwhile, we found 90% of CYP2C19*1/*1 and 10% of CYP2C19*1/*17 in control group. In this study, we identified the frequency of CYP2C19*17 allele in Thai population which will support the pharmacogenetics biomarkers for screening and avoid ADRs of medical cannabis treatment.

Keywords: CYP2C19, allele frequency, ultra rapid metabolizer, medical cannabis

Procedia PDF Downloads 111