Search results for: fenugreek seed extract

Authors: Chengliang Xie, Jinhyun Ryu, Hyun Joon Kim, Gyeong Jae Cho, Wan Sung Choi, Sang Soo Kang, Kye Man Cho, Dong Hoon Lee

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Obesity has become a global health problem and a source of major metabolic diseases like type-2 diabetes, hypertension, heart disease, nonalcoholic fatty liver and cancer. Synthetic anti-obesity drugs are effective but very costly and with undesirable side effects, so natural products such as soybean are needed as an alternative for obesity treatment. Lactobacillus Plantarum P1201is a probiotic bacterial strain reported to produce conjugated linoleic acid (CLA) and increase the ratio of aglycone-isoflavone of soybean, both of which have anti-obesity effect. In this study, the anti-obesity effect of the fermented soybean extract with P1201 (FSE) will be evaluated compared with that of the soybean extract (SE) by 3T3-L1 cells as an in vitro model of adipogenesis. 3T3-L1 cells were treated with SE and FSE during the nine days of the differentiation, lipid accumulation was evaluated by oil-red staining and triglyceride content and the mRNA expression level of adipogenic or lipogenic genes were analyzed by RT-PCR and qPCR. The results showed that formation of lipid droplets in differentiated 3T3-L1 cells was inhibited and triglyceride content was reduced by 23.1% after treated with 1000 μg/mL of FSE compared with control. For SE-treated groups, no delipidating effect was observed. The effect of FSE on adipogenesis inhibition can be attributed to the down-regulation of mRNA expressionof CCAAT/enhancer binding protein (C/EBP-α), lipoprotein lipase (LPL), adiponectin, adipocyte fatty acid-binding protein (aP2), fatty acid synthesis (FAS) and CoA carboxylase (ACC). Our results demonstrated that the anti-obesity effect of soybean can be improved by fermentation with P1201, and P1201can be used as a potential probiotic bacterial strain to produce natural anti-obesity food.

Keywords: fermentation, Lactobacillus plantarum P1201, obesity, soybean

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Authors: Kahlouche Amal, Touzene F. Zohra, Betatache Fatihaet Nouani Abdelouahab

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The development of the world production of the cheese these last decades, as well as agents' greater request cheap coagulants, accentuated the search for new surrogates of the rennet. What about the interest to explore the vegetable biodiversity, the source well cheap of many naturals metabolites that the scientists today praise it (thistle, latex of fig tree, Cardoon, seeds of melon). Indeed, a big interest is concerned the search for surrogates of vegetable origin. The objective of the study is to show the possibility of extracting a protease coagulant the milk from the cake of Sunflower, available raw material and the potential source of surrogates of rennet. so, the determination of the proteolytic activity of raw extracts, the purification, the elimination of the pigments of tint of the enzymatic preparations, a better knowledge of the coagulative properties through study of the effect of certain factors (temperature, pH, concentration in CaCl2) are so many factors which contribute to value milk particularly those produced by the small ruminants of the Algerian dairy exploitations. Otherwise, extracts coagulants of vegetable origin allowed today to value traditional, in addition, although the extract coagulants of vegetable origin made it possible today to develop traditional cheeses whose Iberian peninsula is the promoter, but the test of 'pressed paste not cooked' cheese manufacturing led to the semi-scale pilot; and that, by using the enzymatic extract of sunflower (Helianthus annus) which gave satisfactory results as well to the level of outputs as on the sensory level,which, statistically,did not give any significant difference between studied cheeses. These results confirm the possibility of use of this coagulase as a substitute of rennet commercial on an industrial scale.

Keywords: characterization, cheese, Rennet, sunflower

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Authors: Shraddha Chaudhary, Raksha Agarwal, Niladri Chatterjee

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This work demonstrates a web crawler-based generalized end-to-end open domain Question Answering (QA) system. An efficient QA system requires a significant amount of domain knowledge to answer any question with the aim to find an exact and correct answer in the form of a number, a noun, a short phrase, or a brief piece of text for the user's questions. Analysis of the question, searching the relevant document, and choosing an answer are three important steps in a QA system. This work uses a web scraper (Beautiful Soup) to extract K-documents from the web. The value of K can be calibrated on the basis of a trade-off between time and accuracy. This is followed by a passage ranking process using the MS-Marco dataset trained on 500K queries to extract the most relevant text passage, to shorten the lengthy documents. Further, a QA system is used to extract the answers from the shortened documents based on the query and return the top 3 answers. For evaluation of such systems, accuracy is judged by the exact match between predicted answers and gold answers. But automatic evaluation methods fail due to the linguistic ambiguities inherent in the questions. Moreover, reference answers are often not exhaustive or are out of date. Hence correct answers predicted by the system are often judged incorrect according to the automated metrics. One such scenario arises from the original Google Natural Question (GNQ) dataset which was collected and made available in the year 2016. Use of any such dataset proves to be inefficient with respect to any questions that have time-varying answers. For illustration, if the query is where will be the next Olympics? Gold Answer for the above query as given in the GNQ dataset is “Tokyo”. Since the dataset was collected in the year 2016, and the next Olympics after 2016 were in 2020 that was in Tokyo which is absolutely correct. But if the same question is asked in 2022 then the answer is “Paris, 2024”. Consequently, any evaluation based on the GNQ dataset will be incorrect. Such erroneous predictions are usually given to human evaluators for further validation which is quite expensive and time-consuming. To address this erroneous evaluation, the present work proposes an automated approach for evaluating time-dependent question-answer pairs. In particular, it proposes a metric using the current timestamp along with top-n predicted answers from a given QA system. To test the proposed approach GNQ dataset has been used and the system achieved an accuracy of 78% for a test dataset comprising 100 QA pairs. This test data was automatically extracted using an analysis-based approach from 10K QA pairs of the GNQ dataset. The results obtained are encouraging. The proposed technique appears to have the possibility of developing into a useful scheme for gathering precise, reliable, and specific information in a real-time and efficient manner. Our subsequent experiments will be guided towards establishing the efficacy of the above system for a larger set of time-dependent QA pairs.

Keywords: web-based information retrieval, open domain question answering system, time-varying QA, QA evaluation

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Authors: Nikat Parveen, M. Ananthi

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Big data is one of the emerging technology, which collects the data from various sensors and those data will be used in many fields. Data retrieval is one of the major issue where there is a need to extract the exact data as per the need. In this paper, large amount of data set is processed by using the feature selection. Feature selection helps to choose the data which are actually needed to process and execute the task. The key value is the one which helps to point out exact data available in the storage space. Here the available data is streamed and R-Center is proposed to achieve this task.

Keywords: big data, key value, feature selection, retrieval, performance

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Authors: M. Nuhu, M. S. Amina, A. H. Aminu, A. J. Abbas, N. Salahudeen, A. Z. Yusuf

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Transesterification of jatropha methyl ester (JME) with the common polyol, trimethylolpropane (TMP) produced the TMP based ester which exhibits improved temperature properties. This paper discusses the physic-chemical properties of jatropha bio-lubricant base oil applicable for ISO VG32 and VG46 requirement. The catalyst employed for the JME was CaO synthesized in National Research Institute for Chemical Technology (NARICT) that gives 100% conversion. The molar ratio of JME to TMP was 3.5:1 and the catalyst (NaOCH3) loading were found to be 0.8% of the weight of the total reactants. The final fractionated jatropha bio-lubricant base was found to contain 11.95% monoesters, 43.89% diesters and 44.16% triesters (desired product). In addition, it was found that the bio-lubricant base oil produced is comparable to the ISO VG46 commercial standards for light and industrial gears applications and other plant based bio-lubricant.

Keywords: biodegradability, methyl ester, pour point, transesterification, viscosity index

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Authors: Adeleke G. E., Bello M. A., Abdulateef R. B., Olasinde T. T., Oriaje K. O., AransiI A., Elaigwu K. O., Omidoyin O. S., Shoyinka E. D., Awoyomi M. B., Akano M., Adaramoye O. A.

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Hymenocardia acidatul belongs to the genus, Hymenocardiaceae, which is widely distributed in Africa. Both the leaf and stem bark of the plant have been used in the treatment of several diseases. The present study examined the chemical constituents of the H. acida stem bark extract (HASBE) and its effects on some antioxidant indices and esterase enzymes in female Wistar rats. The HASBE was obtained by Soxhlet extraction using methanol and then subjected to Atomic Absorption Spectroscopy (AAS) for elemental analysis, and Fourier-Transform Infrared (FT-IR) spectroscopy, ultraviolet (UV) spectroscopy, for functional group analysis, while High-performance liquid chromatography (HPLC), and Gas Chromatography-Flame ionization detection (GC-FID) were carried out for compound identification. Forty-eight female Wistar rats were assigned into eight groups of six rats each and separately administered orally with normal saline (Control), 50, 100, 150, 200, 250, 300, 350 mg/kg of HASBE twice per week for eight weeks. The rats were sacrificed under chloroform anesthesia, and kidneys and heart were excised and processed to obtain homogenates. The levels of superoxide dismutase (SOD), catalase, Malondialdehyde (MDA), glutathione peroxidase (GPx), acetylcholinesterase (AChE), and carboxylesterase (CE) were determined spectrophotometrically. The AAS of HASBE shows the presence of eight elements, including Cobalt (0.303), Copper (0.222), Zinc (0.137), Iron (2.027), Nickel (1.304), Chromium (0.313), Manganese (0.213), and Magnesium (0.337 ppm). The FT-IR result of HASBE shows four peaks at 2961.4, 2926.0, 1056.7, and 1034.3 cm-1, while UV analysis shows a maximum absorbance (0.522) at 205 nm. The HPLC spectrum of HASBE indicates the presence of four major compounds, including orientin (77%), β-sitosterol (6.58%), rutin (5.02%), and betulinic acid (3.33%), while GC-FID result shows five major compounds, including rutin (53.27%), orientin (13.06%) and stigmasterol (11.73%), hymenocardine (6.43%) and homopterocarpin (5.29%). The SOD activity was significantly (p < 0.05) lowered in the kidney but elevated in the heart, while catalase was elevated in both organs relative to control rats. The GPx activity was significantly elevated only in the kidney, while MDA was not significantly (p > 0.05) affected in the two organs compared with controls. The activity of AChE was significantly elevated in both organs, while CE activity was elevated only in the kidney relative to control rats. The present study reveals that Hymenocardia acida stem bark extract majorly contains orientin, rutin, stigmasterol, hymenocardine, β-sitosterol, homopterocarpin, and betulinic acid. In addition, these compounds could possibly enhance redox status and esterase activities in the kidney and heart of Wistar rats.

Keywords: hymenocardia acida, elemental analysis, compounds identification, redox status, organs

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Authors: Odunayo Christy Atewolara-Odule, Olapeju O. Aiyelaagbe

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The search for new natural anti-oxidants has grown tremendously over the years because reactive oxygen species (ROS) production and oxidative stress have been linked to a large number of human degenerative diseases, such as cancer, cardiovascular diseases, inflammation, and diabetes. Tapinanthus bangwensis, a parasitic plant commonly known as mistletoe belonging to the Loranthaceae family, is mostly employed traditionally to treat inflammation, cancer, diabetes, and hypertension to mention a few. In this study, air-dried pulverized leaves and stem of Tapinanthus bangwensis were successively extracted with n-hexane, ethyl acetate, and methanol to give the corresponding crude extracts. The extracts were purified by column chromatography and high-performance liquid chromatography to give the isolated compounds. Structural elucidation was done using mass spectrometry, Fourier transform infra-red, 1D and 2D NMR spectroscopy. The antioxidant activity of the compounds was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ascorbic acid as standard. Three compounds; Friedelin, Eudesmic acid (3,4,5-trimethoxybenzoic) and Methyl-3,4,5-trimethoxybenzoate were isolated from the extracts of Tapinanthus bangwensis. Friedelin was isolated from the ethyl acetate extract of the stem while the two other compounds were isolated from the methanol extract of the leaves. The percentages of free radical scavenging activities of the compounds are as follows: Friedelin, 73.69%, methyl-3,4,5-trimethoxybenzoate, 79.33% and eudesmic, 87.68% anti-oxidant activity which were quite comparable to 93.96% given by ascorbic acid. We are reporting, to our best knowledge, for the first time the occurrence of friedelin and eudesmic acid in Tapinanthus bangwensis. The high anti-oxidant activity of these compounds supports the use of this plant in the management of diabetes and hypertension as they will be useful in combating complications arising from the disease.

Keywords: column chromatography, eudesmic acid, friedelin, Tapinanthus bangwensis

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Authors: Narin Salehiyan, Ramin Ghasemi Shayan

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The largest genus of plant viruses, the potyvirus, is responsible for significant crop losses. Potyviruses are aphid sent in a nonpersistent way, and some of them are likewise seed communicated. As significant microorganisms, potyviruses are substantially more examined than other plant infections having a place with different genera, and their review covers numerous parts of plant virology, like utilitarian portrayal of viral proteins, sub-atomic communication with hosts and vectors, structure, scientific classification, development, the study of disease transmission, and determination. Biotechnological utilizations of potyviruses are likewise being investigated. During this last ten years, significant advances have been made in the comprehension of the sub-atomic science of these infections and the elements of their different proteins. Potyvirus multiplication, movement, and transmission, as well as potyvirus/plant compatible interactions, including pathogenicity and symptom determinants, are updated following a general overview of the family Potyviridae and the potyviral proteins. it end the survey giving data on biotechnological uses of potyviruses.

Keywords: virology, poty, virus, genome, genetic

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Authors: Hichem Sebai, Mohamed Amine Jabri, Kais Rtibi, Haifa Tounsi, Lamjed Marzouki

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Artemisia campestris has been widely used in Tunisian traditional medicine for its health beneficial effects. However, the present study aims at evaluating the antiulcer effects of Artemisia campestris aqueous extract (ACAE) as well as the mechanism of action involved in such gastroprotection. In this respect, male Wistar rats were divided into seven groups: control, aspirin (ASPR), ASPR + various doses of ACAE (100, 200 and 400 mg/kg, b.w.), ASPR+ famotidine and ASPR+ caffeic acid. Animals were pre-treated with ACAE extract during 10 days. We firstly showed that aspirin administration was accompanied by an oxidative stress status assessed by an increase of malondialdehyde (MDA) level, a decrease of sulfhydryl -(SH) groups content and depletion of antioxidant enzyme activities such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Pre-treatment with ACAE protected against aspirin-induced gastric oxidative stress. More importantly, aspirin administration increased plasma and tissue hydrogen peroxide (H₂O₂), free iron and calcium levels while the ACAE pre-treatment reversed all aspirin-induced intracellular mediators disturbance. The results of the present study clearly indicated that AEAC gastroprotection might be related, at least in part, to its antioxidant properties as well as to various gastric mucosal defense mechanisms, including the protection of gastric sulfhydryls and an opposite effect on some intracellular mediators such as free iron, hydrogen peroxide, and calcium. However, our data confirm the use of Artemisia campestris extracts in the Tunisian traditional folk medicine for the treatment of gastrointestinal diseases.

Keywords: gastric ulcer, Artemisia campestris, oxidative stress, sulfhydryl groups, Fenton reaction, rat

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Authors: Sandeep Goyal, Sandeep Saluja

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Natural antioxidants have major role in maintenance of health. Green tea extract principally contains epigallocatechin-3-gallate (EGCG), as its abundant antioxidant constituent. Green tea is consumed daily worldwide as antioxidant to combat CNS diseases and has traditional importance also. EGCG has neuroprotective potential in various animal models of Parkinson disease, Alzheimer’s disease etc. but its exact mechanism has not been ruled out. The present study has been designed to investigate the anti-inflammatory, antioxidant and mitochondrial modulating mechanism of neuroprotective effect of epigallocatechin-3-gallate against rodent model of rotenone induced Parkinson’s disease (PD). The behavioural alterations were assessed by using open field test apparatus, Chatilon’s grip strength test apparatus and elevated plus maze for determining the locomotor activity, grip strength and cognition respectively. Biochemically, various parameters to assess oxidative stress, neuroinflammation and neurochemical estimations were performed on rat brain homogenates. A histological examination of rat brain striatum was done to check the neurodegeneration. Epigallocatechin-3-gallate (EGCG) at 10 & 20 mg/kg, were investigated for their neuroprotective potential along with levodopa as a standard agent. Minocycline, a microglial activation inhibitor, was administered alone and in combination with EGCG. EGCG and minocycline produced ameliorative effect against rotenone induced PD like symptoms by significantly reduced behavioral, biochemical and histological alterations. Results of our study reveal the neuroprotective effect of EGCG and minocycline against rotenone induced PD. Results of our study indicate that EGCG exerted neuroprotective effect against rotenone induced PD via its antioxidant, anti-inflammatory and mitochondrial modulating mechanisms and substantiate its previously reported and traditional claims for its use in CNS diseases.

Keywords: antioxidants, neurotoxicity, rotenone, EGCG

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Authors: Uraporn Vongvatcharanon, Kochakorn Sukjan, Wandee Udomuksorn, Ekkasit Kumarnsit, Surapong Vongvatcharanon

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Sarcopenia is a loss of muscle mass, and strength frequently found in menopause. Estrogen replacement has been shown to improve such a loss of muscle functions. However, there is an increased risk of cancer that has to be considered because of the estrogen replacement therapy. Thus, phytoestrogen supplementation has been suggested as an alternative therapy. Pueraria mirifica (PM) is a plant in the family Leguminosae, that is known to be phytoestrogen-rich and has been traditionally used for the treatment of menopausal symptoms. It contains isoflavones and other compounds such as miroestrol and its derivatives. Parvalbumin (PV) is a calcium binding protein and functions as a relaxing factor in fast twitch muscle fibers. A decrease of the PV level results in a reduction of the speed of the twitch relaxation. Therefore, this study aimed to investigate the effect of an ethanolic extract from Pueraria mirifica on the estrogen levels, skeletal muscle functions and PV levels in the extensor digitorum longus (EDL) and gastrocnemius of ovariectomized rats. Twelve-week old female Wistar rats (200-250 g) were divided into 6 groups: SHAM (un-ovariectomized rats, that received double distilled water), PM-0 (ovariectomized rats, OVX, receiving double distilled water), E (OVX, receiving an estradiol benzoate dose of 0.04 mg/kg), PM-50 (OVX receiving PM 50 mg/kg), PM-500 (OVX receiving PM 500 mg/kg), PM-1000 (OVX receiving PM 1000 mg/kg) all for 90 days. The PM-0 group had estrogen levels, uterus weights, muscle mass, myofiber cross-section areas, peak tension, fatigue resistance, speed of relaxation and parvalbumin levels of both EDL and gastrocnemius that were significantly reduced compared to those of the SHAM group (p<0.05). Also the α and β estrogen receptor immunoreactivities and the parvalbumin immunoreactivities of both EDL and gastrocnemius were decreased in the PM-0 group. In contrast the E, PM-50, PM-500 and PM-1000 group had estrogen levels, uterus weights, muscle mass, myofiber cross-section areas, peak tension, fatigue resistance, speed of relaxation of both EDL and gastrocnemius that were significantly increased compared with PM-0 group (p<0.05). In addition, the α and β estrogen receptor immunoreactivities and parvalbumin immunoreactivity of both the EDL and gastrocnemius were increased in the E, PM-50, PM-500 and PM-1000 group. In addition the extract of Pueraria mirifica replacement group at 50 and 500 mg/kg had significantly increased parvalbumin levels in the EDL muscle but in the gastrocnemius, only the dose of 500 mg/kg increased the parvalbumin levels (p<0.05). These results have demonstrated that the use of the Pueraria mirifica extract as a replacement therapy for estrogen produced estrogenic activity that was similar to that produced by the estradiol benzoate replacement. It seems that the phytoestrogens could bind with the estrogen receptors and stimulate the transcriptional activity to synthesise muscle protein that caused an increase in muscle mass and parvalbumin levels. Thus, muscle synthesis may restore parvalbumin levels resulting in an enhanced relaxation efficiency that would lead to a shortened latent period before the next contraction.

Keywords: Puereria mirifica, Parvalbumin, estrogen, ovariectomized rats

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Authors: Narmeen Mehmood

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The current study was conducted to evaluate the pharmacognostic parameters, phytochemical analysis and antibacterial activity of the plant. Microscopic studies were carried out to determine various Pharmacognostic parameters. Section cutting of the leaf was also done. The study of the ariel parts of Beaumontia grandiflora resulted in the identification of fatty acids mixture and unsaponifiable matters. For the separation of various constituents of the plant, successive solvent extraction was carried out in a laboratory. Material and Methods: The study was carried out with all three extracts of Beaumontia grandiflora i.e. Petroleum ether, Chloroform and Methanol. For the separation of various constituents of the plant, successive solvent extraction was carried out in the laboratory. Raw data containing the measured zones of inhibition in mm was tabulated. Results: The microscopic studies showed the presence of Upper epidermis in surface view, Part of Lamina in section view, cortical parenchyma in longitudinal view, Parenchyma with collapsed tissues, Parenchyma Cells, Epidermal cells with a part of covering trichome, starch granules, reticulated thickened vessels, Transverse Section of leaf of Beaumontia grandiflora showed Upper Epidermis, Lower Epidermis, Hairs, Vascular Bundles, Parenchyma. Phytochemical analysis of leaves of Beaumontia grandiflora indicates that Alkaloids are present. There is a possibility of the presence of some bioactive components in the crude extracts due to which it shows strong activity. Petroleum ether extract shows a greater zone of inhibition at low concentrations. Conclusion: The alkaloids possess good antibacterial activity so the presence of alkaloids may be responsible for the antibacterial activity observed in the crude organic extract of Beaumontia grandiflora.

Keywords: successive solvent extraction, zone of inhibitions., microscopy, phytochemical analysis

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Authors: Thomas Q. Pan, Anika Basu, Chamith S. Rajapakse

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Machine learning requires data that is categorized into features that models train on. This topic is important to the field of sports medicine due to the many tools it provides to physicians such as diagnosis support and risk assessment. Physician note that healthcare professionals take are usually unclean and not suitable for model training. The objective of this study was to develop and evaluate an advanced approach for extracting key features from sports medicine data without the need for extensive model training or data labeling. An LLM (Large Language Model) was given a narrative (Physician’s Notes) and prompted to extract four features (details about the patient). The narrative was found in a datasheet that contained six columns: Case Number, Validation Age, Validation Gender, Validation Diagnosis, Validation Body Part, and Narrative. The validation columns represent the accurate responses that the LLM attempts to output. With the given narrative, the LLM would output its response and extract the age, gender, diagnosis, and injured body part with each category taking up one line. The output would then be cleaned, matched, and added to new columns containing the extracted responses. Five ways of checking the accuracy were used: unclear count, substring comparison, LLM comparison, LLM re-check, and hand-evaluation. The unclear count essentially represented the extractions the LLM missed. This can be also understood as the recall score ([total - false negatives] over total). The rest of these correspond to the precision score ([total - false positives] over total). Substring comparison evaluated the validation (X) and extracted (Y) columns’ likeness by checking if X’s results were a substring of Y's findings and vice versa. LLM comparison directly asked an LLM if the X and Y’s results were similar. LLM Re-check prompted the LLM to see if the extracted results can be found in the narrative. Lastly, A selection of 1,000 random narratives was also selected and hand-evaluated to give an estimate of how well the LLM-based feature extraction model performed. With a selection of 10,000 narratives, the LLM-based approach had a recall score of roughly 98%. However, the precision scores of the substring comparison and LLM comparison models were around 72% and 76% respectively. The reason for these low figures is due to the minute differences between answers. For example, the ‘chest’ is a part of the ‘upper trunk’ however, these models cannot detect that. On the other hand, the LLM re-check and subset of hand-tested narratives showed a precision score of 96% and 95%. If this subset is used to extrapolate the possible outcome of the whole 10,000 narratives, the LLM-based approach would be strong in both precision and recall. These results indicated that an LLM-based feature extraction model could be a useful way for medical data in sports to be collected and analyzed by machine learning models. Wide use of this method could potentially increase the availability of data thus improving machine learning algorithms and supporting doctors with more enhanced tools.

Keywords: AI, LLM, ML, sports

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Authors: Benjaporn Buranrat, Nootchanat Mairuae

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Cratoxylum formosum (Jack) Dyer (CF) has been used for the traditional medicines in South East Asian and Thailand. Normally, northeast Thai vegetables have proven cytotoxic to many cancer cells. Therefore, the present study aims to explore the molecular mechanisms underlying CF-induced cancer cell death and apoptosis on breast and liver cancer cells. The cytotoxicity and antiproliferative effects of CF on the human breast MCF-7 and liver HepG2 cancer cell lines were evaluated using sulforhodamine B assay and colony formation assay. Cell migration assay was measured using wound healing assay. The apoptosis induction mechanisms were investigated through reactive oxygen species formation, caspase 3 activity, and JC-1 activity. Gene expression by real-time PCR and apoptosis related protein levels by Western blot analysis. CF induced MCF-7 and HepG2 cell death by time- and dose-dependent manner. Furthermore, CF had the greater cytotoxic potency on MCF-7 more than HepG2 cells with IC50 values of 85.70+4.52 μM and 219.03±9.96 μM respectively, at 24 h. Treatment with CF also caused a dose-dependent decrease in colony forming ability and cell migration, especially on MCF-7 cells. CF induced ROS formation, increased caspase 3 activities, and decreased the mitochondrial membrane potential, and causing apoptotic body production and DNA fragmentation. CF significantly decreased expression of the cell cycle regulatory protein RAC1 and downstream proteins, cdk6. Additionally, CF enhanced p21 and reduced cyclin D1 protein levels. CF leaf extract induced cell death, apoptosis, antimigration in both of MCF-7 and HepG2 cells. CF could be useful for developing to anticancer drug candidate for breast and liver cancer therapy.

Keywords: cratoxylum formosum (jack) dyer, breast cancer, liver cancer, cell death

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Authors: Sineenat Kempubpha, Phornpa-Ngan Muadmuang, Putthamas Phetmuangprab, Surin Promphet, Sopita Bandit

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Introduction: Discarded pericarp of mangosteen (Garcinia mangostana) is a benefit to be developed as veterinary phytopharmacal products since it made up of abundance pharmacological active compounds. The active compounds of mangosteen pericarp not only act as an antihistamine, an anti-inflammatory, heart disease and HIV therapeutic substances but also act as antibacterial and antifungal agents. Aim: This study was an in vitro procedural attempt to determine the antibacterial effects of mangosteen pericarp 95% ethanol extract on the main causative pathogen of canine superficial pyoderma, Staphylococcus pseudintermedius. Methods: S. pseudintermedius were collected from various sites of the skin of canine superficial pyoderma dogs and were revived and lawn cultured. The S. pseudintermedius growth inhibition study was determined by disc diffusion technique, the mangosteen pericarp crude extracted was dissolved in 3 types of solvents (95% ethanol, 2% DMSO and distilled water, respectively). The micro broth dilution technique was used for determining both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. Statistical analysis was done by calculating the mean of the zones of inhibition of tested microorganisms. Results: S. pseudintermedius growth inhibition study showed that the inhibition efficacy of 95% ethanol was greater than the inhibition efficacy of 2% DMSO and distilled water (9.10±0.18 mm, 6.95±0.60 mm and 6.80±0.18 mm, respectively). The MIC value was 125 µg/ml and the MBC value was 1 mg/ml. Conclusion: Mangosteen pericarp extract dissolved with 95% ethanol showed the highest zone of inhibition against the tested microorganisms. The MIC value was 125 µg/ml and the MBC value was 1 mg/ml which suggests its potent antibacterial action against S. pseudintermedius. However, further analytical studies are needed to isolate the key molecules of mangosteen pericarp for higher effect on canine superficial pyoderma microorganism therapeutic products.

Keywords: mangosteen, Garcinia mangostana, Staphylococcus pseudintermedius, canine superficial pyoderma, in vitro study

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Authors: Rafik Balti, Mohamed Ben Mansour, Abdellah Arhaliass, Anthony Masse

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Unfortunately, shrimps are highly perishable and they start deteriorating immediately after death owing to their high water content and nutritional components. Currently, there has been an increasing interest in bioactive edible films and coatings to preserve the freshness and quality of foods. In this study, active edible coatings from microalgal exopolysaccharides (EPS) enriched with different concentrations of Red Seaweed Extract (RSE) (0.5, 1 and 1.5 % (w/v)) were developed and their effects on the quality changes of white shrimp during refrigerated storage (4 ± 1 °C) were examined over a period of 8 days. The control and the coated shrimp samples were analyzed periodically for microbiological (total viable bacteria, psychrotrophic bacteria, and enterobacteriaceae counts), chemical (pH, TVB-N, TMA-N, PV, TBARS), textural and sensory characteristics. The results indicated that the coating with a mixture of EPS and RSE could significantly decrease the total volatile basic nitrogen (TVB-N), trimethylamine (TMA) and thiobarbituric acid reactive substances (TBARS) (p < 0.05). With storage, EPS coatings containing RSE at both levels (1 and 1.5 %) were more effective in inhibiting the microbial species studied, specially psychrotrophic bacteria. Also, EPS + RSE coated samples had lower polyphenol oxidase (PPO) activity and lipid oxidation (p < 0.05) toward the end of storage. Textural and color properties of coated shrimp were generally more acceptable. Sensory scores indicated no significant changes in all samples during storage. The obtained results indicate that the edible EPS coating solutions enriched with RSE have noticeable effects on the quality and shelf life of shrimps when compared to control group. Finally, the present work demonstrates the effectiveness of EPS enriched coatings, offering a promising alternative to preserve more better the quality characteristics and to extend the shelf life of shrimp during the refrigerated storage

Keywords: active coating, exopolysaccharides, red seaweed, refrigerated storage, white shrimp

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Authors: Hadjer Didouh, Mohammed Hadj Melliani, Izzeddine Sameut Bouhaik

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Corrosion is a serious problem in industrial installations or metallic transport pipes. Corrosion is an interfacial process controlled by several parameters. The presence of microorganisms affects the kinetics of corrosion. This type of corrosion is often referred to as bio-corrosion or corrosion influenced by microorganisms (MIC). The action of a microorganism or a bacterium is carried out by the formation of biofilm following its attachment to the metal surface. The formation of biofilm isolates the metal surface from its environment and allows the bacteria to control the parameters of the metal/bacteria interface. Biofilm formation by sulfate-reducing bacteria (SRB) X52 steel poses substantial challenges in the oil and gas industry SONATRACH of Algeria. This research delves into the complex attachment mechanisms employed by SRB biofilm on X52 carbon steel and investigates innovative strategies for effective mitigation using biocides. The exploration commences by elucidating the underlying mechanisms facilitating SRB biofilm adhesion to X52 carbon steel, considering factors such as surface morphology, electrostatic interactions, and microbial extracellular substances. Advanced microscopy and spectroscopic techniques provide support to the attachment processes, laying the foundation for targeted mitigation strategies. The use of 100 ppm of Moringa Oleifera extract biocide as a promising approach to control and prevent SRB biofilm formation on X52 carbon steel surfaces. Green extracts undergo evaluation for their effectiveness in disrupting biofilm development while ensuring the integrity of the steel substrate. Systematic analysis is conducted on the biocide's impact on the biofilm's structural integrity, microbial viability, and overall attachment strength. This two-pronged investigation aims to deepen our comprehension of SRB biofilm dynamics and contribute to the development of effective strategies for mitigating its impact on X52 carbon steel.

Keywords: attachment, bio-corrosion, biofilm, metal/bacteria interface

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Authors: Rita Chattopadhyay, Vivek Anand Thoutam

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Speech is the most acceptable means of communication where we can quickly exchange our feelings and thoughts. Quite often, people can communicate orally but cannot interact or work with computers or devices. It’s easy and quick to give speech commands than typing commands to computers. In the same way, it’s easy listening to audio played from a device than extract output from computers or devices. Especially with Robotics being an emerging market with applications in warehouses, the hospitality industry, consumer electronics, assistive technology, etc., speech-based human-machine interaction is emerging as a lucrative feature for robot manufacturers. Considering this factor, the objective of this paper is to design the “Audio-Visual Co-Data Processing Pipeline.” This pipeline is an integrated version of Automatic speech recognition, a Natural language model for text understanding, object detection, and text-to-speech modules. There are many Deep Learning models for each type of the modules mentioned above, but OpenVINO Model Zoo models are used because the OpenVINO toolkit covers both computer vision and non-computer vision workloads across Intel hardware and maximizes performance, and accelerates application development. A speech command is given as input that has information about target objects to be detected and start and end times to extract the required interval from the video. Speech is converted to text using the Automatic speech recognition QuartzNet model. The summary is extracted from text using a natural language model Generative Pre-Trained Transformer-3 (GPT-3). Based on the summary, essential frames from the video are extracted, and the You Only Look Once (YOLO) object detection model detects You Only Look Once (YOLO) objects on these extracted frames. Frame numbers that have target objects (specified objects in the speech command) are saved as text. Finally, this text (frame numbers) is converted to speech using text to speech model and will be played from the device. This project is developed for 80 You Only Look Once (YOLO) labels, and the user can extract frames based on only one or two target labels. This pipeline can be extended for more than two target labels easily by making appropriate changes in the object detection module. This project is developed for four different speech command formats by including sample examples in the prompt used by Generative Pre-Trained Transformer-3 (GPT-3) model. Based on user preference, one can come up with a new speech command format by including some examples of the respective format in the prompt used by the Generative Pre-Trained Transformer-3 (GPT-3) model. This pipeline can be used in many projects like human-machine interface, human-robot interaction, and surveillance through speech commands. All object detection projects can be upgraded using this pipeline so that one can give speech commands and output is played from the device.

Keywords: OpenVINO, automatic speech recognition, natural language processing, object detection, text to speech

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Authors: Gary Pattemore, Michael Edgar, Andrew Job, Marina Auad, Kathryn Job

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As critical commodities become more scarce, significant time and resources have been used to better understand complicated ore bodies and extract their full potential. These challenging ore bodies provide several pain points for geologists and engineers to overcome, poor handling of these issues flows downs stream to the processing plant affecting throughput rates and recovery. Many open cut mines utilise blast hole drilling to extract additional information to feed back into the modelling process. This method requires samples to be collected during or after blast hole drilling. Samples are then sent for assay with turnaround times varying from 1 to 12 days. This method is time consuming, costly, requires human exposure on the bench and collects elemental data only. To address this challenge, research has been undertaken to utilise hyperspectral imaging across a broad spectrum to scan samples, collars or take down hole measurements for minerals and moisture content and grade abundances. Automation of this process using unmanned vehicles and on-board processing reduces human in pit exposure to ensure ongoing safety. On-board processing allows data to be integrated into modelling workflows with immediacy. The preliminary results demonstrate numerous direct and indirect benefits from this new technology, including rapid and accurate grade estimates, moisture content and mineralogy. These benefits allow for faster geo modelling updates, better informed mine scheduling and improved downstream blending and processing practices. The paper presents recommendations for implementation of the technology in open cut mining environments.

Keywords: grade control, hyperspectral scanning, artificial intelligence, autonomous mining, machine learning

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Authors: Alise Senberga, Laila Dubova, Liene Strauta, Ina Alsina, Ieva Erdberga

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Legume symbiotic relationship with nitrogen fixating bacteria Rhizobim leguminosarum is an important factor used to improve the productivity of legumes, due to the fact that rhizobia can supply plant with the necessary amount of nitrogen. R. leguminosarum strains have shown different activity in fixing nitrogen. Depending on the chosen R. leguminosarum strain, host plant biochemical content can be altered. In this study we focused particularly on the changes in protein content in beans (using two different varieties) and peas (five different varieties) due to the use of several different R. leguminosarum strains (four strains for both beans and peas). Overall, the protein content increase was observed after seed inoculation with R. leguminosarum. Strain and plant cultivar interaction specification was observed. The effect of R. leguminosarum inoculation on the content of protein was dependent on the R. leguminosarum strain used. Plant cultivar also appeared to have a decisive role in protein content formation with the help of R. leguminosaru.

Keywords: legumes, protein content, rhizobia strains, soil

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Authors: A. Adetutu, T. M. Awodugba, O. A. Owoade

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The development of resistance to currently known conventional anti-typhoid drugs has necessitated search into cheap, more potent and less toxic anti-typhoid drugs of plant origin. Therefore, this study investigated the anti-typhoid activity of fractions of A. indica in Salmonella typhi infected rats. Leaves of A. indica were extracted in methanol and fractionated into n-hexane, chloroform, ethyl-acetate, and aqueous fractions. The anti-salmonella potentials of fractions of A. indica were assessed via in-vitro inhibition of S. typhi using agar well diffusion, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and biofilm assays. The biochemical and haematological parameters were determined by spectrophotometric methods. The histological analysis was performed using Haematoxylin and Eosin staining methods. Data analysis was performed by one-way ANOVA. Results of this study showed that S. typhi was sensitive to aqueous and chloroform fractions of A. indica, and the fractions showed biofilm inhibition at concentrations of 12.50, 1.562, and 0.39 mg/mL. In the in-vivo study, the extract and chloroform fraction had significant (p < 0.05) effects on the number of viable S. typhi recovered from the blood and stopped salmonellosis after 6 days of treatment of rats at 500 mg/kg b.w. Treatments of infected rats with chloroform and aqueous fractions of A. indica normalized the haematological parameters in the animals. Similarly, treatment with fractions of the plants sustained a normal antioxidant status when compared with the normal control group. Chloroform and ethyl-acetate fractions of A. indica reversed the liver and intestinal degeneration induced by S. typhi infection in rats. The present investigation indicated that the aqueous and chloroform fractions of A. indica showed the potential to provide an effective treatment for salmonellosis, including typhoid fever. The results of the study may justify the ethno-medicinal use of the extract in traditional medicine for the treatment of typhoid and salmonella infections.

Keywords: Azadirachta indica L, salmonella, typhoid, leave fractions

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Authors: Juan Alfredo Guevara Carrio, Swamy Toolahalli Thipperudra, Riddhi Naik Dharmeshbhai, Sergio Graniero Echeverrigaray, Jose Vitorio Emiliano, Antonio Helio Castro

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In order to advance the hydrogen economy, several industrial sectors can potentially benefit from the trillions of stimulus spending for post-coronavirus. Blending hydrogen into natural gas pipeline networks has been proposed as a means of delivering it during the early market development phase, using separation and purification technologies downstream to extract the pure H₂ close to the point of end-use. This first step has been mentioned around the world as an opportunity to use existing infrastructures for immediate decarbonisation pathways. Among current technologies used to extract hydrogen from mixtures in pipelines or liquid carriers, membrane separation can achieve the highest selectivity. The most efficient approach for the separation of H₂ from other substances by membranes is offered from the research of 2D layered materials due to their exceptional physical and chemical properties. Graphene-based membranes, with their distribution of pore sizes in nanometers and angstrom range, have shown fundamental and economic advantages over other materials. Their combination with the structure of ceramic and geopolymeric materials enabled the synthesis of nanocomposites and the fabrication of membranes with long-term stability and robustness in a relevant range of physical and chemical conditions. Versatile separation modules have been developed for hydrogen separation, which adaptability allows their integration in industrial prototypes for applications in heavy transport, steel, and cement production, as well as small installations at end-user stations of pipeline networks. The developed membranes and prototypes are a practical contribution to the technological challenge of supply pure H₂ for the mentioned industries as well as hydrogen energy-based fuel cells.

Keywords: graphene nano-composite membranes, hydrogen separation and purification, separation modules, indsutrial prototype

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Authors: Rishabh Beri, Sahil Shah

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We have applied a range of filters and processing in order to extract out the various features of the football game, like the field lines of a football field. Another important aspect was the detection of the players in the field and tagging them according to their teams distinguished by their jersey colours. This extracted information combined about the players and field helped us to create a virtual field that consists of the playing field and the players mapped to their locations in it.

Keywords: Detect, Football, Players, Virtual

Procedia PDF Downloads 334

Authors: Prerana Sharma

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Effect of relativistic nonlinearity on stimulated Raman scattering of the propagating laser beam carrying null intensity in center (hollow Gaussian beam) by excited plasma wave are studied in a collisionless plasma. The construction of the equations is done employing the fluid theory which is developed with partial differential equation and Maxwell’s equations. The analysis is done using eikonal method. The phenonmenon of Stimulated Raman scattering is shown along with the excitation of seed plasma wave. The power of plasma wave and back reflectivity is observed for higher order of hollow Gaussian beam. Back reflectivity is studied numerically for various orders of HGLB with different value of plasma density, laser power and beam radius. Numerical analysis shows that these parameters play vital role on reflectivity characteristics.

Keywords: Hollow Gaussian beam, relativistic nonlinearity, plasma physics, Raman scattering

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Authors: Victor Pena Ribeiro, Caroline Arruda, Jennyfer Andrea Aldana Mejia, Jairo Kenupp Bastos

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Leishmaniasis is a serious health problem around the world. The treatment of infected individuals with pentavalent antimonial drugs is the main therapeutic strategy. However, they present high toxicity and persistence side effects. Therefore, the discovery of new and safe natural-derived therapeutic agents against leishmaniasis is important. Propolis is a resin of viscous consistency produced by Apis mellifera bees from parts of plants. The main types of Brazilian propolis are green, red, yellow and brown. Thus, the aim of this work was to investigate the chemical composition and leishmanicidal properties of a brown propolis (BP). For this purpose, the hydroalcoholic crude extract of BP was obtained and was fractionated by liquid-liquid chromatography. The chemical profile of the extract and its fractions were obtained by HPLC-UV-DAD. The fractions were submitted to preparative HPLC chromatography for isolation of the major compounds of each fraction. They were analyzed by NMR for structural determination. The volatile compounds were obtained by hydrodistillation and identified by GC/MS. Promastigote forms of Leishmania amazonensis were cultivated in M199 medium and then 2×106 parasites.mL-1 were incubated in 96-well microtiter plates with the samples. The BP was dissolved in dimethyl sulfoxide (DMSO) and diluted into the medium, to give final concentrations of 1.56, 3.12, 6.25, 12.5, 25 and 50 µg.mL⁻¹. The plates were incubated at 25ºC for 24 h, and the lysis percentage was determined by using a Neubauer chamber. The bioassays were performed in triplicate, using a medium with 0.5% DMSO as a negative control and amphotericin B as a positive control. The leishimnicidal effect against promastigote forms was also evaluated at the same concentrations. Cytotoxicity experiments also were performed in 96-well plates against normal (CHO-k1) and tumor cell lines (AGP01 and HeLa) using XTT colorimetric method. Phenolic compounds, flavonoids, and terpenoids were identified in brown propolis. The major compounds were identified as follows: p-coumaric acid (24.6%) for a methanolic fraction, Artepelin-C (29.2%) for ethyl acetate fraction and the compounds of hexane fraction are in the process of structural elucidation. The major volatile compounds identified were β-caryophyllene (10.9%), germacrene D (9.7%), nerolidol (10.8%) and spathulenol (8.5%). The propolis did not show cytotoxicity against normal cell lines (CHO) with IC₅₀ > 100 μg.mL⁻¹, whereas the IC₅₀ < 10 μg.mL⁻¹ showed a potential against the AGP01 cell line, propolis did not demonstrate cytotoxicity against HeLa cell lines IC₅₀ > 100 μg.mL⁻¹. In the determination of the leishmanicidal activity, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations of the crude extract caused the lysis of 76% and 45% of promastigote forms of L. amazonensis, respectively. To the amastigote form, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations caused the mortality of 89% and 75% of L. amazonensis, respectively. The IC₅₀ was 2.8 μg.mL⁻¹ to amastigote form and 3.9 μg.mL⁻¹ to promastigote form, showing a promising activity against Leishmania amazonensis.

Keywords: amastigote, brown propolis, cytotoxicity, promastigote

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Authors: Velasco-Rendón María Del Carmen, Cuéllar-Bermúdez Sara Paulina, Parra-Saldívar Roberto

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Phycocyanin is a blue pigment protein with fluorescent activity produced by cyanobacteria. It has been recently studied to determine its anticancer, antioxidant and antiinflamatory potential. Since 2014 it was approved as a Generally Recognized As Safe (GRAS) proteic pigment for the food industry. Therefore, phycocyanin shows potential for the food, nutraceutical, pharmaceutical and diagnostics industry. Conventional phycocyanin extraction includes buffer solutions and ammonium sulphate followed by chromatography or ATPS for protein separation. Therefore, further purification steps are time-requiring, energy intensive and not suitable for scale-up processing. This work presents an alternative to conventional methods that also allows large scale application with commercially available equipment. The extraction was performed by exposing the dry biomass to mechanical cavitation and salting out with NaCl to use an edible reagent. Also, isoelectric point precipitation was used by addition of HCl and neutralization with NaOH. The results were measured and compared in phycocyanin concentration, purity and extraction yield. Results showed that the best extraction condition was the extraction by salting out with 0.20 M NaCl after 30 minutes cavitation, with a concentration in the supernatant of 2.22 mg/ml, a purity of 3.28 and recovery from crude extract of 81.27%. Mechanical cavitation presumably increased the solvent-biomass contact, making the crude extract visibly dark blue after centrifugation. Compared to other systems, our process has less purification steps, similar concentrations in the phycocyanin-rich fraction and higher purity. The contaminants present in our process edible NaCl or low pHs that can be neutralized. It also can be adapted to a semi-continuous process with commercially available equipment. This characteristics make this process an appealing alternative for phycocyanin extraction as a pigment for the food industry.

Keywords: extraction, phycocyanin, precipitation, scale-up

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Authors: Ali Fadhel Ahmed, Tuqa Abdulkareem Hameed

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Moringa oleifera (leaves and seeds) ethanolic and aqueous extracts were tested for antibacterial activity. The effect of plant extracts on three types of bacterial species: Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, was investigated. Using the agar well diffusion method, ethanolic extracts of Moringa oleifera demonstrated a significant antibacterial effect on the forty tested bacterial strains. Seed-induced inhibition zones (ethanolic extracts)were ranged from16 to 24 mm in diameter against S. aureus, respectively, whileE. coli and K. pneumonia had no effect. Gram-positive and Gram-negative bacteria were not affected by alcoholic and aqueous plant leaf extracts. The purpose of this present study was to look at the cytotoxic effects of M.Oleifera plant (alcoholic extracts).

Keywords: moringa oleifera, escherichia coli, klebsiella pneumoniae, staphylococcus aureus

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Authors: Rosa Palmeri, Julieta I. Monteleone, Antonio C. Barbera, Carmelo Maucieri, Aldo Todaro, Virgilio Giannone, Giovanni Spagna

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In recent years, considerable interest has been shown in the functional properties of foods, and a relevant role has been played by phenolic compounds, able to scavenge free radicals. A more sustainable agriculture has to emerge to guarantee food supply over the next years. Wheat, corn, and rice are the most common cereals cultivated, but also other cereal species, such as barley can be appreciated for their peculiarities. Barley (Hordeum vulgare L.) is a C3 winter cereal that shows high resistance at drought and salt stresses. There are growing interests in barley as ingredient for the production of functional foods due to its high content of phenolic compounds and Beta-glucans. In this respect, the possibility of separating specific functional fractions from food industry by-products looks very promising. Olive leaves represent a quantitatively significant by-product of olive grove farming, and are an interesting source of phenolic compounds. In particular, oleuropein, which provide important nutritional benefits, is the main phenolic compound in olive leaves and ranges from 17% to 23% depending upon the cultivar and growing season period. Together with oleuropein and its derivatives (e.g. dimethyloleuropein, oleuropein diglucoside), olive leaves further contain tyrosol, hydroxytyrosol, and a series of secondary metabolities structurally related to them: verbascoside, ligstroside, hydroxytyrosol glucoside, tyrosol glucoside, oleuroside, oleoside-11-methyl ester, and nuzhenide. Several flavonoids, flavonoid glycosides, and phenolic acids have also described in olive leaves. The aim of this work was the production of functional food with higher content of polyphenols and the evaluation of their shelf life. Organic durum wheat and barley grains contain higher levels of phenolic compounds were used for the production of crackers. Olive leaf extract (OLE) was obtained from cv. ‘Biancolilla’ by aqueous extraction method. Two baked goods trials were performed with both organic durum wheat and barley flours, adding olive leaf extract. Control crackers, made as comparison, were produced with the same formulation replacing OLE with water. Total phenolic compound, moisture content, activity water, and textural properties at different time of storage were determined to evaluate the shelf-life of the products. Our the preliminary results showed that the enriched crackers showed higher phenolic content and antioxidant activity than control. Alternative uses of olive leaf extracts for crackers production could represent a good candidate for the addition of functional ingredients because bakery items are daily consumed, and have long shelf-life.

Keywords: barley, functional foods, olive leaf, polyphenols, shelf life

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Authors: Sali Chinsathit, Haruthai Kaenla

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

Keywords: banana, longkong, mangosteen, organic farming

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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