Search results for: animal protein concentrate

Authors: V. Konjik, J. Schwartz, R. Sandhoff, M. Mack

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The rising number of multi-resistant pathogens demands the development of new antibiotics in order to reduce the lethal risk of infections. Here, we investigate roseoflavin, a vitamin B2 analogue which is produced by Streptomyces davawensis and Streptomyces cinnabarinus. We consider roseoflavin to be a 'Trojan horse' compound. Its chemical structure is very similar to riboflavin but in fact it is a toxin. Furthermore, it is a clever strategy with regard to the delivery of an antibiotic to its site of action but also with regard to the production of this chemical: The producer cell has only to convert a vitamin (which is already present in the cytoplasm) into a vitamin analog. Roseoflavin inhibits the activity of Flavin depending proteins, which makes up to 3.5 % of predicted proteins in organisms sequenced so far. We sequentially knocked out gene clusters and later on single genes in order to find the ones which are involved in the roseoflavin biosynthesis. Consequently, we identified the gene rosB, coding for the protein carrying out the first step of roseoflavin biosynthesis, starting form Flavin mononucleotide. Here we show, that the protein RosB has so far unknown features. It is per se an oxidoreductase, a decarboxylase and an aminotransferase, all rolled into one enzyme. A screen of cofactors revealed needs of oxygen, NAD+, thiamine and glutamic acid to carry out its function. Surprisingly, thiamine is not only needed for the decaboxylation step, but also for the oxidation of 8-demethyl-8-formyl Flavin mononucleotide. We had managed to isolate three different Flavin intermediates with different oxidation states, which gave us a mechanistic insight of RosB functionality. Our work points to a so far new function of thiamine in Streptomyces davawensis. Additionally, RosB could be extremely useful for chemical synthesis. Careful engineering of RosB may allow the site-specific replacement of methyl groups by amino groups in polyaromatic compounds of commercial interest. Finally, the complete clarification of the roseoflavin biosynthesis opens the possibility of engineering cost-effective roseoflavin producing strains.

Keywords: antibiotic, flavin analogue, roseoflavin biosynthesis, vitamin B2

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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: Xinxiu Li, Chuqian Zheng, Yanyan Qian, Hong Fan

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Objectives: In hepatocellular carcinoma (HCC), oncogenes are continuously and robustly transcribed due to aberrant expression of essential components of the trans-acting super-enhancers (SE) complex. Preclinical and clinical trials are now being conducted on small-molecule inhibitors that target core-transcriptional components, including as transcriptional bromodomain protein 4 (BRD4) and cyclin-dependent kinase 7 (CDK7), in a number of malignant tumors. This study aims to explore whether co-overexpression of BRD4 and CDK7 is a potential marker of worse prognosis and a combined therapeutic target in HCC. Methods: The expression pattern of BRD4 and CDK7 and their correlation with prognosis in HCC were analyzed by RNA sequencing data and survival data of HCC patients from TCGA and GEO datasets. The protein levels of BRD4 and CDK7 were determined by immunohistochemistry (IHC), and survival data of patients were analyzed using the Kaplan-Meier method. The mRNA expression levels of genes in HCC cell lines were evaluated by quantitative PCR (q-PCR). CCK-8 and colony formation assays were conducted to assess cell proliferation of HCC upon treatment with BRD4 inhibitor JQ1 or/and CDK7 inhibitor THZ1. Results: It was shown that BRD4 and CDK7 were often overexpressed in HCCs and were associated with poor prognosis of HCC by analyzing the TCGA and GEO datasets. BRD4 or CDK7 overexpression was related to a lower survival rate. It's interesting to note that co-overexpression of CDK7 and BRD4 was a worse prognostic factor in HCC. Treatment with JQ1 or THZ1 alone had an inhibitory effect on cell proliferation; however, when JQ1 and THZ1 were combined, there was a more notable suppression of cell growth. At the same time, the combined use of JQ1 and THZ1 synergistically suppresses the expression of HCC driver genes. Conclusion: Our research revealed that BRD4 and CDK7 coupled can be a useful biomarker in HCC prognosis and the combination of JQ1 and THZ1 can be a promising therapeutic therapy against HCC.

Keywords: BRD4, CDK7, cell proliferation, combined inhibition

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Authors: Iñaki Milton-Laskibar, Leixuri Aguirre, Maria P. Portillo

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Introduction: Energy restriction is an effective approach in preventing liver steatosis. However, due to social and economic reasons among others, compliance with this treatment protocol is often very poor, especially in the long term. Resveratrol, a natural polyphenolic compound that belongs to stilbene group, has been widely reported to imitate the effects of energy restriction. Objective: To analyze the effects of resveratrol under normoenergetic feeding conditions and under a mild energy restriction on liver fat accumulation and hepatic fatty acid oxidation. Methods: 36 male six-week-old rats were fed a high-fat high-sucrose diet for 6 weeks in order to induce steatosis. Then, rats were divided into four groups and fed a standard diet for 6 additional weeks: control group (C), resveratrol group (RSV, resveratrol 30 mg/kg/d), restricted group (R, 15 % energy restriction) and combined group (RR, 15 % energy restriction and resveratrol 30 mg/kg/d). Liver triacylglycerols (TG) and total cholesterol contents were measured by using commercial kits. Carnitine palmitoyl transferase 1a (CPT 1a) and citrate synthase (CS) activities were measured spectrophotometrically. TFAM (mitochondrial transcription factor A) and peroxisome proliferator-activator receptor alpha (PPARα) protein contents, as well as the ratio acetylated peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α)/Total PGC1α were analyzed by Western blot. Statistical analysis was performed by using one way ANOVA and Newman-Keuls as post-hoc test. Results: No differences were observed among the four groups regarding liver weight and cholesterol content, but the three treated groups showed reduced TG when compared to the control group, being the restricted groups the ones showing the lowest values (with no differences between them). Higher CPT 1a and CS activities were observed in the groups supplemented with resveratrol (RSV and RR), with no difference between them. The acetylated PGC1α /total PGC1α ratio was lower in the treated groups (RSV, R and RR) than in the control group, with no differences among them. As far as TFAM protein expression is concerned, only the RR group reached a higher value. Finally, no changes were observed in PPARα protein expression. Conclusions: Resveratrol administration is an effective intervention for liver triacylglycerol content reduction, but a mild energy restriction is even more effective. The mechanisms of action of these two strategies are different. Thus resveratrol, but not energy restriction, seems to act by increasing fatty acid oxidation, although mitochondriogenesis seems not to be induced. When both treatments (resveratrol administration and a mild energy restriction) were combined, no additive or synergic effects were appreciated. Acknowledgements: MINECO-FEDER (AGL2015-65719-R), Basque Government (IT-572-13), University of the Basque Country (ELDUNANOTEK UFI11/32), Institut of Health Carlos III (CIBERobn). Iñaki Milton is a fellowship from the Basque Government.

Keywords: energy restriction, fat, liver, oxidation, resveratrol

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Authors: Lorena Coelho, David Ramada, Catarina Nobre, Joaquim Gaião, Juliana Duarte

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The development of processes that allows the valorization of waste and by-products generated by industries is crucial to promote symbiotic relationships between different sectors and is mandatory to “close the loop” in the circular economy paradigm. In recent years, by-products and waste from agro-food and forestry sector have attracted attention due to their potential application and technical characteristics. The extraction of bio-based active compounds to be reused is in line with the circular bioeconomy concept trends, combining the use of renewable resources with the process’s circularity, aiming the waste reduction and encouraging reuse and recycling. Among different types of bio-based materials, which are being explored and can be extracted, proteins fractions are becoming an attractive new raw material. Within this context, BioTrace4Leather project, a collaboration between two Technological Centres – CeNTI and CTIC, and a company of Tanning and Finishing of Leather – Curtumes Aveneda, aims to develop innovative and biologically sustainable solutions for leather industry and accomplish the market circularity trends. Specifically, it aims to the valorisation of waste and by-products from the tannery industry through proteins extraction and the development of an innovative and biologically sustainable materials. The achieved results show that keratin, gelatine, and collagen fractions can be successfully extracted from hair and leather bovine waste. These products could be reintegrated into the industrial manufacturing process to attain innovative and functional textile and leather substrates. ACKNOWLEDGEMENT This work has been developed under BioTrace4Leather scope, a project co-funded by Operational Program for Competitiveness and Internationalization (COMPETE) of PORTUGAL2020, through the European Regional Development Fund (ERDF), under grant agreement Nº POCI-01-0247-FEDER-039867.

Keywords: leather by-products, circular economy, sustainability, protein fractions

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Authors: Wisdom Robert Duruji, Oluwasegun Christopher Akinleye

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This study reports on the impact of different ripening accelerators on the microbial load and proximate composition of plantain (Musa paradisiaca) and Banana (Musa sapientum) during the ripening process, and the nutrition implication for food security. The study comprised of four treatments, namely: Calcium carbide, Irvingia gabonensis fruits, Newbouldia laevis leaves and a control, where no ripening accelerator was applied to the fingers of plantain and banana. The unripe and ripened plantain and banana were subjected to microbial analysis by isolating and enumerating their micro flora using pour plate method; and also, their proximate composition was determined using standard methods. The result indicated that the bacteria count of plantain increased from 3.25 ± 0.33 for unripe to 5.31 ± 0.30 log cfu/g for (treated) ripened, and that of banana increased from 3.69 ± 0.11 for unripe to 5.26 ± 0.21 log cfu/g for ripened. Also, the fungal count of plantain increased from 3.20 ± 0.16 for unripe to 4.88 ± 0.22 log sfu/g for ripened; and that of banana increased from 3.61 ± 0.19 for unripe to 5.43 ± 0.26 for ripened. Ripened plantain fingers without any ripening accelerator (control) had significantly (p < 0.05) higher values of crude protein 3.56 ± 0.06%, crude fat 0.42 ± 0.04%, total ash 2.74 ± 0.15 and carbohydrate 31.10 ± 0.20; but with significantly lower value of moisture 62.14 ± 0.07% when compared with treated plantain. The proximate composition trend of treated and banana fingers control is similar to that of treated and plantain control, except that higher moisture content of 75.11 ± 0.07% and lesser protein, crude fat, total ash and carbohydrate were obtained from treated and ripened banana control when the treatments were compared with that of plantain. The study concluded that plantain is more nutritious (mealy) than a banana; also, the ripening accelerators increased the microbial load and reduced the nutritional status of plantain and banana.

Keywords: food nutrition, calcium carbide, rvingia gabonensis, newbouldia laevis, plantain, banana

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Authors: Chengalappa Seenappa, Narayanappa Devkumar, Narayanappa Nagaraja

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Mulberry leaf is the major economic component in sericulture and quality of leaf produced per unit area has a direct effect on quality of cocoon. Among all the agronomical inputs, irrigation water has highest impact on mulberry leaf quantity and quality. The water productivity in sericulture in the country is inadequate and inefficient though India has the largest irrigated area. There is a need of proper irrigation methods and conservation practices to ensure efficiency and economy in water use. Hence, this field experiment was conducted at College of Sericulture, Chintamani, Chickaballapur district, Karnataka, India during 2013 and 2014 to know the quantity, quality and water productivity of mulberry influenced by different methods, levels of irrigation and mulching in Eastern Dry Zone (EDZ) of Karnataka, India. The results revealed that the mulberry leaf quantity, quality and water productivity were significantly influenced by different methods, levels of irrigation and mulching. Subsurface drip irrigation at 0.8 CPE (Cumulative Pan Evaporation) recorded higher leaf yield, chlorophyll, relative water, protein content and water productivity (42857 kg ha-1 yr-1, 8.54, 65.80%, 22.27% and 364.41 kg hacm-1, respectively) than surface drip at 1.0 CPE (38809 kg ha-1 yr-1, 7.34, 62.76%, 17.75% and 264 10 kg hacm-1, respectively) and micro spray jet at 1.0 CPE (39931 kg ha-1 yr-1, 7.96, 63.50%, 19.00%, 35617 kg ha-1 yr-1 and 271.83 kg hacm-1, respectively). Mulching treatment recorded maximum leaf yield, chlorophyll, relative water, protein content and water productivity (38035 kg ha-1 yr-1, 7.12, 62.11%, 16.14% and 330 kg hacm-1, respectively) compared to without mulching. These results clearly indicated that subsurface drip irrigation at lower level of irrigation (0.8 CPE) and mulching increased the quantity, quality and water productivity of mulberry leaf than surface drip and micro spray jet irrigation at higher level of irrigation (1.0 CPE) by saving 20 per cent of water. Therefore, in the coming days subsurface drip irrigation in mulberry cultivation may be more appropriate to realise higher yield, quality and water productivity in EDZ of Karnataka, India.

Keywords: subsurface drip irrigation, mulching, water productivity, mulberry

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Authors: Musawira Iftikhar

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The field of membrane materials is the most dynamic due to the constantly evolving requirements advancement of materials, to address challenges such as biocompatibility, protein-bound uremic toxins, blood coagulation, auto-immune responses, oxidative stress, and poor clearance of uremic toxins. Hemodialysis is a membrane filtration processes that is currently necessary for daily living of the patients with ESRD. Tens of millions of people with ESRD have benefited from hemodialysis over the past 60–70 years, both in terms of safeguarding life and a longer lifespan. Beyond challenges associated with the efficiency and separative properties of the membranes, ensuring hemocompatibility, or the safe circulation of blood outside the body for four hours every two days, remains a persistent challenge. This review explores the ongoing field of metal–Organic Frameworks (MOFs) and their applications in hemodialysis, offering a comprehensive examination of various MOFs employed to address challenges inherent in traditional hemodialysis methodologies. this This review included includes the experimental work done with various MOFs as a filler such as UiO-66, HKUST-1, MIL-101, and ZIF-8, which together lead to improved adsorption capacities for a range of uremic toxins and proteins. Furthermore, this review highlights how effectively MOF-based hemodialysis membranes remove a variety of uremic toxins, including p-cresol, urea, creatinine, and indoxyl sulfate and potential filler choices for the future. Future research efforts should focus on refining synthesis techniques, enhancing toxin selectivity, and investigating the long-term durability of MOF-based membranes. With these considerations, MOFs emerge as transformative materials in the quest to develop advanced and efficient hemodialysis technologies, holding the promise to significantly enhance patient outcomes and redefine the landscape of renal therapy.

Keywords: membrane, hemodailysis, metal organic frameworks, seperation, protein adsorbtion

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Authors: Ülle Parm, Kata Pedamäe, Jaak Jürimäe, Evelin Lätt, Aivar Orav, Anna-Liisa Tamm

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Vegetarianism has gained popularity across the world. It`s being chosen for multiple reasons, but among Estonians, these have remained unknown. Previously, attention to bone health and probable nutrient deficiency of vegetarians has been paid and in vegetarians lower body mass index (BMI) and blood cholesterol level has been found but the results are inconclusive. The goal was to explain reasons for choosing vegetarian diet in Estonia and impact of vegetarianism to body composition – BMI, fat percentage (fat%), fat mass (FM), and fat free mass (FFM). The study group comprised of 68 vegetarians and 103 omnivorous. The determining body composition with DXA (Hologic) was concluded in 2013. Body mass (medical electronic scale, A&D Instruments, Abingdon, UK) and height (Martin metal anthropometer to the nearest 0.1 cm) were measured and BMI calculated (kg/m2). General data (physical activity level included) was collected with questionnaires. The main reasons why vegetarianism was chosen were the healthiness of the vegetarian diet (59%) and the wish to fight for animal rights (72%) Food additives were consumed by less than half of vegetarians, more often by men. Vegetarians had lower BMI than omnivores, especially amongst men. Based on BMI classification, vegetarians were less obese than omnivores. However, there were no differences in the FM, FFM and fat percentage figures of the two groups. Higher BMI might be the cause of higher physical activity level among omnivores compared with vegetarians. For classifying people as underweight, normal weight, overweight and obese both BMI and fat% criteria were used. By BMI classification in comparison with fat%, more people in the normal weight group were considered; by using fat% in comparison with BMI classification, however, more people categorized as overweight. It can be concluded that the main reasons for vegetarianism chosen in Estonia are healthiness of the vegetarian diet and the wish to fight for animal rights and vegetarian diet has no effect on body fat percentage, FM and FFM.

Keywords: body composition, body fat percentage, body mass index, vegetarianism

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

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Background: Breast cancer is emerging as one of the leading cause of cancer related deaths and hence there arises the need to look out for drugs which are more targets specific with minimal side effects. In recent times, there is a shift towards alternative medicine due to low cost and less side effects. Siddha system of medicine is one the oldest system of medicine practiced against various ailments. Tridham (TD) is a herbal formulation prepared in our laboratory consisting of Terminalia chebula, Elaeocarpus ganitrus and Prosopis cineraria in a definite ratio (TD) and its anticancer potential is evaluated in terms of induction of apoptosis. Objective: The present study was designed to investigate the anti proliferative effect of TD and 1,2,3,4,6-penta-O-galloyl-b-D-glucose (PGG), a pure compound isolated from TD on human mammary carcinoma cell line (MCF-7). Materials and Methods: Cell viability was studied using MTT analysis and trypan blue staining. Mitochondrial membrane potential was studied using DAPI staining. The protein and mRNA expressions of pro-apoptotic and anti- apoptotic markers namely Bax, Bad, Bcl-2 and caspases were also assessed by Western Blotting and RT PCR. Results: Viability studies of TD and PGG treated MCF-7 cells showed an inhibition in cell growth in time and dose dependent manner. The alteration in mitochondrial membrane potential was restored through treatment with TD and PGG which was confirmed by DAPI staining. The protein and mRNA expression of pro-apoptotic markers was found to be significantly increased in TD and PGG treated cells with a concomitant decrease in anti-apoptotic markers. Conclusion: The results of the study suggest that TD and PGG exhibit their anticancer effect through its membrane stabilizing property and activation of apoptotic cascade in MCF-7 cells.

Keywords: apoptosis, mammary carcinoma, MCF-7, penta galloyl glucose, Tridham

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Authors: Iahtasham Khan, Vania Lucia De Assis Santana, Marcilia Maria Alves De Souza, Mabel Hanna Vance Harrop, Fernando Leandro Dos Santos, Cecília Maria Souza Leão E. Silva, Pedro Paulo Silveira, Marcelo Brasil, Marcus Vinícius, Hélio Cordeiro Manso Filho, Muhammad Younus, Aman Ullah Khan

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Glanders ranks high on clinical lists in some regions of Brazil as a cause of respiratory and lymphatic disease in equids. Glanders is caused by Burkolderia mallei (B. mallei) Gram-negative bacterium. B. mallei was first biological agent used in World War I in 20th century. The complement fixation test (CFT) is a serodiagnostic tool prescribed by the World Organization for Animal Health (OIE)for the diagnosis of glanders in the international trade of equids. The aim of the present study was to monitor the serological responses in equines naturally infected with B. mallei using the CFT. A total of 574 equids were tested with CFT, 30 days apart in a total of 12 samplings. One hundred thirty-four sera tested negative in all samplings; 192 sera tested positive in one sampling and 125 sera tested positive in two or more samplings. Remaining 123 samples showed uncertain results. Thus, CFT results can vary over a period of time. These variations could be the consequence of the effects of the natural immune response in each animal. The findings of the present study demonstrate difficulties regarding the simultaneous implementation of CFT and test and slaughter policies to eradicate glanders. Another constraint to control this disease is the presence of carrier/transitory CFT-negative animals, which are a potential source of disease in glanders-free areas. Serodiagnostic tests of higher sensitivity and specificity like immunobloat should be implemented to achieve success in the eradication of glanders.

Keywords: glanders, equids, horses, immunological, mules

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Authors: Madelyn N. Moawad, Hermine R. Z. Tadros, Mary G. Ghobrial, Ahmad R. Bassiouny, Kamal M. Kandeel, Athar Ata

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Seaweeds are potential renewable resources in marine environment. They provide an excellent source of bioactive substances such as dietary fibers and various functional polysaccharides that could potentially be used as ingredients for both human and animal health applications. The observations suggested that these bioactive compounds have strong antioxidant properties, which have beneficial effects on human health. The present research aimed at finding new chemical products from local marine macroalgae for natural medicinal uses and consumption for their nutritional values. Macroalgae samples were collected manually mainly from the Mediterranean Sea at shallow subtidal zone of Abu Qir Bay, Alexandria, Egypt. The chemical compositions of lyophilized materials of six selected macroalgal species; Colpomenia sinuosa, Sargassum linifolium, Padina pavonia, Pterocladiella capillacea, Laurencia pinnatifidia, and Caulerpa racemosa, were investigated for proteins using bovine serum albumin, and carbohydrates were assayed by phenol-sulfuric acid reaction. The macroalgae lipid was extracted with chloroform, methanol and phosphate buffer. Vitamins were extracted using trichloroacetic acid. Chlorophylls and total carotenoids were determined spectrophotometrically and total phenols were extracted with methanol. In addition, lipid-soluble, and water-soluble antioxidant, and anti α-glucosidase activities were measured spectrophotometrically. The antioxidant activity of hexane extracts was investigated using phosphomolybdenum reagent. The anti-α-glucosidase effect measurement was initiated by mixing α-glucosidase solution with p-nitrophenyl α-D-glucopyranoside. The results showed that the ash contents varied from 11.2 to 35.4 % on dry weight basis for P. capillacea and Laurencia pinnatifidia, respectively. The protein contents ranged from 5.63 % in brown macroalgae C. sinuosa to 8.73 % in P. pavonia. A relative wide range in carbohydrate contents was observed (20.06–46.75 %) for the test algal species. The highest lipid percentage was found in green alga C. racemosa (5.91%) followed by brown algae P. pavonia (3.57%) and C. sinuosa (2.64%). The phenolic contents varied from 1.32 mg GAE/g for C. sinuosa to 4.00 mg GAE/g in P. pavonia. The lipid-soluble compounds exhibited higher antioxidant capacity (73.18-145.95 µM/g) than that of the water-soluble ones ranging from 24.83 µM/g in C. racemosa to 74.07 µM/g in S. linifolium. The most potent anti-α-glucosidase activity was observed for P. pavonia with IC50 of 17.12 μg/ml followed by S. linifolium (IC50 = 71.75 μg/ml), C. racemosa (IC50 = 84.73 μg/ml), P. capillacea (IC50 = 92.16 μg/ml), C. sinuosa (IC50 = 112.44 μg/ml), and L. pinnatifida (IC50 = 115.11 μg/ml).

Keywords: α-glucosidase, lyophilized, macroalgae, spectrophotometrically

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Authors: Nacer Eddine Chelbi, Ayet Bagane, Annie Saleh, Claude Sauvageau, Denis Gingras

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As recently stated by National Highway Traffic Safety Administration (NHTSA), to demonstrate the expected performance of a highly automated vehicles system, test approaches should include a combination of simulation, test track, and on-road testing. In this paper, we propose a new validation method for autonomous vehicles involving on-road tests (Field Operational Tests), test track (Test Matrix) and simulation (Worst Case Scenarios). We concentrate our discussion on the simulation aspects, in particular, we extend recent work based on Importance Sampling by using a Metropolis-Hasting algorithm (MHS) to sample collected data from the Safety Pilot Model Deployment (SPMD) in lane-change scenarios. Our proposed MH sampling method will be compared to the Importance Sampling method, which does not perform well in high-dimensional problems. The importance of this study is to obtain a sampler that could be applied to high dimensional simulation problems in order to reduce and optimize the number of test scenarios that are necessary for validation and certification of autonomous vehicles.

Keywords: automated driving, autonomous emergency braking (AEB), autonomous vehicles, certification, evaluation, importance sampling, metropolis-hastings sampling, tests

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Authors: Liana C. Salanță, Maria Tofană, Carmen R. Pop, Vlad Mureșan

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The landscape of functional food is expanding very fast, due to the consumer interest for healthy natural products. Consumers nowadays demand healthy products that impart phytonutrients to encourage good health and well-being, prevent diseases, without sacrificing taste and texture. Candies are foodstuffs appreciated by all category of consumers. They are available in a range variety of forms (jellies, marshmallows, caramels, lollipops, etc.). Jelly is characterized by a gummy and chewy texture typically conferred by a hydrocolloid (gelatin, pectin). The purpose of this research was to obtain hypocaloric jelly (no added sugar) enriched with protein powder from acai, chia seeds and hemp, which are considered superfood. Peach and raspberry juice were used for obtaining functional jelly, due to the specific flavour, natural carbohydrate, natural pigments and vitamins (C, B1, PP, etc). Instead of classic hydrocolloids used in Romania for the industry of jelly, agar-agar was used in this study, due to its properties. Agar-agar is able to form gels in the aqueous medium, stronger than other gel-forming agents. High sugar concentrations or an acid environment (as is necessary with pectins) are not needed. In addition to its gelation properties, Agar-agar is considered to have important nutritional benefits, high content of fibre and has low calories. Six prototypes of jellies were obtained and evaluated by physicochemical, microbiological and sensorial analysis. For the textural profile analysis, the Brookfield CT3 Texture Analyzer, equipped with a 10kg load cell, was used. The results revealed that hypocaloric jelly can serve as a good source of bioactive compounds in the diet. The jelly is a convenient way of delivering potential health benefits of protein powder and agar-agar to a wide range of consumers.

Keywords: agar-agar, functional food, hypocaloric jelly, superfoods

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Authors: Roike Iwan Montolalu, Henny Adeleida Dien, Feny Mentang, Kristhina P. Rahael, Tomy Moga, Ayub Meko, Siegfried Berhimpon

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In the last 20 years, packaging materials derived from petrochemicals polymers were widely used as packaging materials. This due to various advantages such as flexible, strong, transparent, and the price is relatively cheap. However, the plastic polymer also has various disadvantages, such as the transmission monomer contamination into the material to be packed, and waste is non-biodegradable. Edible film (EF) is an up to date materials, generated after the biodegradable packaging materials. The advantages of the EF materials, is the materials can be eat together with food, and the materials can be applied as a coating materials for a widely kind of foods especially snack foods. The aims of this research are to produce and to analyze the characteristics of smoked EF made from carrageenan, myofibril and collagen of Black Marlin (Makaira indica) industrial waste. Smoked EF made with an addition of 0.8 % smoke liquid. Three biopolymers i.e. carrageenan, myofibril, and collagen were used as treatments, and homogenate for 1 hours at speed of 1500 rpm. The analysis carried out on the pH and physical properties i.e. thickness, solubility, tensile strength, % elongation, and water vapor transmission rate (WVTR), as well as on the sensory characteristics of texture i.e. wateriness, firmness, elasticity, hardness, and juiciness of the coated products. The result shown that the higher the concentration the higher the thickness of EF, where as for myofibril proteins appeared higher than carrageenan and collagen. Both of collagen and myofibril shown that concentration of 6% was most soluble, while for carrageenan were in concentration of 2 to 2.5%. For tensile strength, carrageenan was significantly higher than myofibril and collagen; while for elongation, collagen film more elastic than carragenan and myofibril protein. Water vapor transmission rate, shown that myofibril protein film lower than carrageenan and collagen film. From sensory assessment of texture, carrageenan has a high elasticity and juiciness, while collagen and myofibril have a high in firmness and hardness.

Keywords: edible film, collagen, myofibril, carrageenan

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Authors: Sajjad Ur Rahman, Muhammad Tahir, Mukarram Bashir, Jawad, Aoun Muhammad, Muhammad Zohaib, Hannan Khan, Seemal Javaid, Mariam Azam

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The efficiency of natural metabolites obtained from partially fermented soya hulls and wheat bran using Saccharomyces cerevisiae (DL-22 S/N) ensures a potential impact on the total milk yield and quality of milk production. On attaining a moderate number of Saccharomyces cerevisiae cells around 1×10⁹ CFU/ml, the concentrate was further elevated under in-vivo conditions to study the quality of milk production in lactating buffalo. Ten lactating buffalos of the Nili Ravi breed having the same physical factors were given 12 gm of microbial metabolites daily, along with the palleted feed having 22 % proteins. Another group of 10 lactating animals with the same characteristics was maintained without metabolites. The body score, overall health, incidence of mastitis, milk fat, milk proteins, ash and solid not fat (SNF) were elevated on a weekly basis up to thirty days of trial. It was recorded that the average total increase in quality milk production was 0.9 liter/h/d, whereas SNF in the milk was enhanced to 0.71, and fats were decreased to 0.09 %. Moreover, during all periods of the trial, the overall non-specific immunity of buffalo was increased, as indicated by less than 0.2 % of mastitis incidence compared to 1.8% in the untreated buffalos.

Keywords: natural metabolites, quality milk, milk yield, microorganisms, fermentation, nonspecific immunity, better performing animals

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Authors: Hyo Jung Lee

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The purpose of this study is to develop a literature appreciation education program for classic literatures and apply them to the field, and to derive the achievements and improvement points. Classic literature is a work of value recognized in the context of literature history and culture history, and learners can develop interest in literature and inherit tradition through appreciation of classic literature. However, in Korean educational environment, classic literature is a means for college entrance examination, and many learners analyze contents and language in textbooks and concentrate on memorizing the whole plot. This study is one of the reasons that classic literature appreciation education is not done properly and it is not able to give an opportunity to appreciate the whole work in the early learning stage. In Korean primary education, classic literature is used as a means to achieve the goals of reading, writing, speaking and listening, rather than being used as a material for its own appreciation. It is problematic to make the piece appreciation experience fragmentary. This study proposes a program to experience classic literatures by linking school education and school library with primary school students in grades 4-6. We work with local primary schools (siheung-si, gyeonggi-do, Korea) to provide appropriate activities and rewards to learners, observe their participation, and introduce student learning outcomes. Through this, we are able to systematically improve the learner 's ability to appreciate the literature and to diversify primary education.

Keywords: classic literature education, primary education, out-of-school program, learning by appreciation experience

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Authors: Sudarshan Kalsulkar, Sunil S. Bhagwat

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Biosorption is a physiochemical process that occurs naturally in certain biomass which allows it to passively concentrate and bind contaminants onto its cellular structure. Activated carbons (AC) are one such efficient biosorbents made by utilizing lignocellulosic materials from agricultural waste. Steam activated carbon (AC) was synthesized from Barley husk. Its synthesis parameters of time and temperature were optimized. Its physico-chemical properties like density, surface area, pore volume, Methylene blue and Iodine values were characterized. BET surface area was found to be 42 m²/g. Batch Adsorption tests were carried out to determine the maximum adsorption capacity (qmax) for various metal ions. Cd+2 48.74 mg/g, Pb+2 19.28 mg/g, Hg+2 39.1mg/g were the respective qmax values. pH and time were optimized for adsorption of each ion. Column Adsorptions were carried for each to obtain breakthrough data. Microbial adsorption was carried using E. coli K12 strain. 78% reduction in cell count was observed at operating conditions. Thus the synthesized Barley husk AC can be an economically feasible replacement for commercially available AC prepared from the costlier coconut shells. Breweries and malting industries where barley husk is a primary waste generated on a large scale can be a good source for bulk raw material.

Keywords: activated carbon, Barley husk, biosorption, decontamination, heavy metal removal, water treatment

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Authors: Wong Seng Yue

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Game dynamics and game mechanics are the two main components that used in gamification to engage and encourage students to learn. The advantages of gamified classroom are engaging students, increasing students interest, preserving students focus and remain a positive behaviour. However, the empirical studies on gamification are still at early stage, especially the effectiveness of various gamification components have not been evaluated. Thus, this study is aimed to conduct a user experience (UX) evaluation on gamified classroom through Prezi, which focused on learning experience, gaming experience, adaptivity, and gameplay experience. This study is a further study extended from the previous exploratory study to explore more on UX of gamified classroom via Prezi by interview. A focus group study, which involves 22 students from a foundation course has been conducted for the study. Besides the empirical data from the previous study, this focus group study has significantly found that 90.9% respondents show their positive perceptions on gaming experience via Prezi. They are interested, feel fresh, good, and highly motivated of the contents of Prezi. 95.5% participants have had a positive learning experience from the gamified classroom via Prezi, which can engage them, made them concentrate on learning and easy to remember what they have learned if compared to the traditional classroom slides. The adaptivity of the gamified classroom also high due to its zooming user interface, narrative, rewards and engagement features. This study has uncovered on how far the impact of gamification components in the classroom, especially UX that implemented in gamified classroom.

Keywords: user experience (UX), gamification, gamified classroom, Prezi

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Authors: Bruno C. Schimming, Leandro L. Martins, PatríCia F. F. Pinheiro, Raquel F. Domeniconi, FabríCio S. Oliveira

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The agouti is a wildlife rodent that can be used as an alternative source of animal protein and this species has been raised in captivity in Brazil with the aim of providing meat. Thus, the knowledge of their reproductive biology and morphology of the reproductive organs is important. The objective of this study was to describe the morphology of epididymis in the Azara’s agouti, by light microscopy. Samples of epididymis were obtained from five adult Azara’s agouti (Dasyprocta azarae) during castration surgery performed at the Municipal Zoo of Catanduva, Brazil. Fragments of the epididymal regions (initial segment, caput, corpus and cauda) were collected. The biological samples were immediately fixed in paraformaldehyde for 24 hours, followed by histologic procedures comprising embedding in ParaplastTM (Sigma, St. Louis, MO, USA), sections of 5 µm, and staining with HE and Masson’s trichrome. The epididymis was a highly convoluted tubule that links the testis to the vas deferens. The epithelium lining was pseudostratified columnar surrounded by a periductal stroma. The epithelium contains several cell types: principal, basal, apical, clear, and hallo cells. Principal cells were the most abundant cell type. There were observed also migratory cells named halo cells. The caput epididymis was divided into two different regions: initial segment and caput. The initial segment has a very wide lumen, a high epithelium with conspicuous microvilli and the lumen was wide with exfoliated material. The other region of the caput epididymis, showed a lower epithelium when compared with the initial segment, large amounts of spermatozoa in the lumen, and a cytoplasmic vacuolization. This region presented many narrows cells. Many spermatozoa appeared in the lumen of corpus epididymis. The cauda region had a lower epithelium than the other epididymal regions in the agouti. The cauda epithelium presented plicae protruding into the lumen. Large amounts of spermatozoa are also present in the lumen. Small microvilli uniformly arranged so as to form a kind of “brush border” are observed on the apical surface of the cauda epithelium. The pattern of the epithelium lining the duct of the agouti epididymis does not differ greatly from that reported to other mammals, such as domestic and wildlife animals. These findings can cooperate with future investigations especially those related to rational exploration of these animals. All experimental procedures were approved by the institutional ethics committee (CEUA 796/2015). This study was supported by FAPESP (Grants 2015/23822-1).

Keywords: wildlife, testis excurrent ducts, epididymis, morphology

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Authors: Klára Gajdošíková

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The aim of this study was to examine the quiet eye method and its components using the mobile eye tracking device. Quiet eye training was proven to be beneficial for different sports, including golf. The main idea of this method is to prolong your fixations on a specific place in order to improve your performance. The shot examined in this study is called putt. Its importance is based on its role on a golf course because, many times, it is the last putt that decides whether you win or lose. Quiet eye training helps players be more focused under pressure, control their attention and overall improve their putting success. Six highly skilled golfers with a handicap ranging from - 4 to + 4, aged 23 to 26, participated in a pilot study with the usage of an eye-tracking device. The study took place in an indoor training area at the golf club Hostivař. Crosstabs showed significant differences between individuals' laterality and their gaze into AOI - areas of interest (middle part of the ball, top of the ball, bottom of the ball, back side of the ball). Statistically significant differences were also discovered between the mean fixation duration of participants with AOI on the middle part of the ball and all other AOIs. The results of this study helped us understand the examined phenomena and showed us the next aim in future quiet eye research. Future research should focus on examining a quiet eye on the golf course. Applying a quiet eye and therefore changing the way we concentrate might be beneficial for coaches and players themselves.

Keywords: eye tracking, golf, laterality, quiet eye

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Authors: Getachew Belay Kassahun

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Background: Locally established clinical laboratory reference intervals (RIs) are required to interpret laboratory test results for screening, diagnosis, and prognosis. The objective of this study was to establish a reference interval of clinical chemistry parameters among apparently healthy adolescents aged between 12 and 17 years in Mekelle, Tigrai, in the northern part of Ethiopia. Methods: Community-based cross-sectional study was employed from December 2018 to March 2019 in Mekelle City among 172 males and 172 females based on a Multi-stage sampling technique. Blood samples were tested for Fasting blood sugar (FBS), alanine amino transferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), Creatinine, urea, total protein, albumin (ALB), direct and indirect bilirubin (BIL.D and BIL.T) using 25 Bio system clinical chemistry analyzer. Results were analyzed using SPSS version 23 software and based on the Clinical Laboratory Standard Institute (CLSI)/ International Federation of Clinical Chemistry (IFCC) C 28-A3 Guideline which defines the reference interval as the 95% central range of 2.5th and 97.5th percentiles. Mann Whitney U test, descriptive statistics and box and whisker were statistical tools used for analysis. Results: This study observed statistically significant differences between males and females in ALP, ALT, AST, Urea and Creatinine Reference intervals. The established reference intervals for males and females, respectively, were: ALP (U/L) 79.48-492.12 versus 63.56-253.34, ALT (U/L) 4.54-23.69 versus 5.1-20.03, AST 15.7- 39.1 versus 13.3- 28.5, Urea (mg/dL) 9.33-24.99 versus 7.43-23.11, and Creatinine (mg/dL) 0.393-0.957 versus 0.301-0.846. The combined RIs for Total Protein (g/dL) were 6.08-7.85, ALB (g/dL) 4.42-5.46, FBS(mg/dL) 65-110, BIL.D (mg/dL) 0.033-0.532, and BIL.T (mg/dL) 0.106-0.812. Conclusions: The result showed a marked difference between sex and company-derived values for selected clinical chemistry parameters. Thus, the use of age and sex-specific locally established reference intervals for clinical chemistry parameters is recommended.

Keywords: reference interval, adolescent, clinical chemistry, Ethiopia

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Authors: Ceyda Okudu, Secil Eroglu, Khandakar A. S. M. Saadat, Sibel O. Balci

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Ring Finger Protein 2 (RNF2) is a member of polycomb repressive complex 1 (PRC1), which is one of the epigenetic regulators in the genome. When RNF2 combines with other PRC1 members, it mediates the mono-ubiquitination of Histon2A (H2A). In breast cancer, RNF2 is commonly overexpressed, and also it promotes metastasis and invasion in other aggressive tumors like melanoma, prostate, and hepatocarcinoma. The role of RNF2 in the metastasis and invasion of breast cancer has not yet been elucidated. Our aim is to observe the role of RNF2 in metastasis and invasion in this study by miRNA mediated RNF2 gene silencing in breast cancer cell lines. We selected miRNAs, targeting to RNF2 by searching online databases. miR-17-5p, miR20a-5p, and miR-106b-5p were transfected to breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, and ZR-75-1), and also we used normal breast epithelial cell line (hTERT-HME1) to compare RNF2 gene expression level. After 48-72 hours post-transfection, mRNAs were isolated from the cells, and gene expressions were measured by RT-qPCR after from cDNA syntheses. We observed that RNF2 was highly expressed in SK-BR-3 and MDA-MB-231 cell lines opposite to MCF-7 and ZR-75-1 cell lines. RNF2 was downregulated 5, 5 and 7 fold by miR17-5p, miR20a-5p and miR106b-5p respectively in MCF-7. However, in SK-BR-3 and ZR-75-1 cell lines, miRNAs did not affect significantly RNF2 gene expression level. miR20a-5p decreased RNF2 3 fold and miR17-5p and miR106b-5p did not affect MDA-MB-231. After gene expression analysis, we performed metastasis and invasion assay in MCF-7 cells. For metastasis, we used both wound healing assay and Transwell Cell Migration Assay, and we used Transwell Cell Invasion Assay for invasion. The data of this assay showed that miR17-5p and miR20a-5p decreased both invasion and metastasis level, but miR106b-5p has no effect. We would like to conclude that RNF2 can be targeted by miR17-5p, miR20a-5p and miR106b-5p in MCF-7 cells and also RNF2, which is one of the upregulated genes in aggressive tumor, can be decreased by using these miRNAs. In future, we would like to confirm these results at the protein level and also whether these miRNAs are direct target of RNF2 or not.

Keywords: breast cancer, epigenetic, microRNAs, RNF2

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Authors: Mauricio Filho, Carina Castro

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The object of this project is to analyze long-time public agents that passed through several governments and see themselves in the position of having to deliberate with new agents, recently settled in the public administration. For theoretical ends, internal deliberation is understood as the one practiced on the public administration agencies, without any direct participation from the general public in the process. The assumption is: agents with longer periods of public service tend to step away from momentary political discussions that guide the current administration and seek to concentrate on institutionalized routines and procedures, making the most politically aligned individuals with the current government deliberate with less "passion" and more exchanging of knowledge and information. The theoretical framework of this research is institutionalism, which is guided by a more pragmatic view, facing the fluidity of reality in ways showing the multiple relations between agents and their respective institutions. The critical aspirations of this project rest on the works of professors Cass Sunstein, Adrian Vermeule, Philipp Pettit and in literature from both institutional theory and economic analysis of law, greatly influenced by the Chicago Law School. Methodologically, the paper is a theoretical review and pretends to be unfolded, in a future moment, in empirical tests for verification. This work has as its main analytical tool the appeal to theoretical and doctrinaire areas from the Juridical Sciences, by adopting the deductive and analytical method.

Keywords: institutions, state, law, agencies

Procedia PDF Downloads 266

Authors: Tomás Sobrino, Lara García-Varela, Marta Aramburu-Núñez, Mónica Castro, Noemí Gómez-Lado, Mariña Rodríguez-Arrizabalaga, Antía Custodia, Juan Manuel Pías-Peleteiro, José Manuel Aldrey, Daniel Romaus-Sanjurjo, Ángeles Almeida, Pablo Aguiar, Alberto Ouro

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Introduction: Alzheimer’s disease (AD) and other tauopathies are primary causes of dementia, causing progressive cognitive deterioration that entails serious repercussions for the patients' performance of daily tasks. Currently, there is no effective approach for the early diagnosis and treatment of AD and tauopathies. This study suggests a theragnostic approach based on the importance of phosphorylated tau protein (p-Tau) in the early pathophysiological processes of AD. We have developed a novel theragnostic monoclonal antibody (mAb) to provide both diagnostic and therapeutic effects. Methods/Results: We have developed a p-Tau mAb, which was doped with deferoxamine for radiolabeling with Zirconium-89 (89Zr) for PET imaging, as well as fluorescence dies for immunofluorescence assays. The p-Tau mAb was evaluated in vitro for toxicity by MTT assay, LDH activity, propidium iodide/Annexin V assay, caspase-3, and mitochondrial membrane potential (MMP) assay in both mouse endothelial cell line (bEnd.3) and cortical primary neurons cell cultures. Importantly, non-toxic effects (up to concentrations of p-Tau mAb greater than 100 ug/mL) were detected. In vivo experiments in the tauopathy model mice (PS19) show that the 89Zr-pTau-mAb and 89Zr-Fragments-pTau-mAb are stable in circulation for up to 10 days without toxic effects. However, only less than 0.2% reached the brain, so further strategies have to be designed for crossing the Brain-Blood-Barrier (BBB). Moreover, an intraparenchymal treatment strategy was carried out. The PS19 mice were operated to implement osmotic pumps (Alzet 1004) at two different times, at 4 and 7 months, to stimulate the controlled release for one month each of the B6 antibody or the IgG1 control antibody. We demonstrated that B6-treated mice maintained their motor and memory abilities significantly compared with IgG1 treatment. In addition, we observed a significant reduction in p-Tau deposits in the brain. Conclusions /Discussion: A theragnostic pTau-mAb was developed. Moreover, we demonstrated that our p-Tau mAb recognizes very-early pathology forms of p-Tau by non-invasive techniques, such as PET. In addition, p-Tau mAb has non-toxic effects, both in vitro and in vivo. Although the p-Tau mAb is stable in circulation, only 0.2% achieve the brain. However, direct intraventricular treatment significantly reduces cognitive impairment in Alzheimer's animal models, as well as the accumulation of toxic p-Tau species.

Keywords: alzheimer's disease, theragnosis, tau, PET, immunotherapy, tauopathies

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Authors: Befekadu Urga Wakayo

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Introduction: Bacterial infections represent major human and animal health problems in Ethiopia. In the face of poor antibiotic regulatory mechanisms, development of antimicrobial resistance (AMR) to commonly used drugs has become a growing health and livelihood threat in the country. Monitoring and control of AMR demand close coloration between human and veterinary services as well as other relevant stakeholders. However, risk of AMR transfer from animal to human population’s remains poorly explored in Ethiopia. This systematic research literature review attempted to give an overview on AMR challenges of bovine mastitis bacteria in Ethiopia. Methodology: A web based research literature search and analysis strategy was used. Databases are considered including; PubMed, Google Scholar, Ethiopian Veterinary Association (EVA) and Ethiopian Society of Animal Production (ESAP). The key search terms and phrases were; Ethiopia, dairy, cattle, mastitis, bacteria isolation, antibiotic sensitivity and antimicrobial resistance. Ultimately, 15 research reports were used for the current analysis. Data extraction was performed using a structured Microsoft Excel format. Frequency AMR prevalence (%) was registered directly or calculated from reported values. Statistical analysis was performed on SPSS – 16. Variables were summarized by giving frequencies (n or %), Mean ± SE and demonstrative box plots. One way ANOVA and independent t test were used to evaluate variations in AMR prevalence estimates (Ln transformed). Statistical significance was determined at p < 0.050). Results: AMR in bovine mastitis bacteria was investigated in a total of 592 in vitro antibiotic sensitivity trials involving 12 different mastitis bacteria (including 1126 Gram positive and 77 Gram negative isolates) and 14 antibiotics. Bovine mastitis bacteria exhibited AMR to most of the antibiotics tested. Gentamycin had the lowest average AMR in both Gram positive (2%) and negative (1.8%) bacteria. Gram negative mastitis bacteria showed higher mean in vitro resistance levels to; Erythromycin (72.6%), Tetracycline (56.65%), Amoxicillin (49.6%), Ampicillin (47.6%), Clindamycin (47.2%) and Penicillin (40.6%). Among Gram positive mastitis bacteria higher mean in vitro resistance was observed in; Ampicillin (32.8%), Amoxicillin (32.6%), Penicillin (24.9%), Streptomycin (20.2%), Penicillinase Resistant Penicillin’s (15.4%) and Tetracycline (14.9%). More specifically, S. aurues exhibited high mean AMR against Penicillin (76.3%) and Ampicillin (70.3%) followed by Amoxicillin (45%), Streptomycin (40.6%), Tetracycline (24.5%) and Clindamycin (23.5%). E. coli showed high mean AMR to Erythromycin (78.7%), Tetracycline (51.5%), Ampicillin (49.25%), Amoxicillin (43.3%), Clindamycin (38.4%) and Penicillin (33.8%). Streptococcus spp. demonstrated higher (p =0.005) mean AMR against Kanamycin (> 20%) and full sensitivity (100%) to Clindamycin. Overall, mean Tetracycline (p = 0.013), Gentamycin (p = 0.001), Polymixin (p = 0.034), Erythromycin (p = 0.011) and Ampicillin (p = 0.009) resistance increased from the 2010’s than the 2000’s. Conclusion; the review indicated a rising AMR challenge among bovine mastitis bacteria in Ethiopia. Corresponding, public health implications demand a deeper, integrated investigation.

Keywords: antimicrobial resistance, dairy cattle, Ethiopia, Mastitis bacteria

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Authors: Joanna Skopinska-Wisniewska, Anna Bajek, Marta Ziegler-Borowska, Alina Sionkowska

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Hydrogels are a class of materials widely used in medicine for many years. Proteins, such as collagen, due to the presence of a large number of functional groups are easily wettable by polar solvents and can create hydrogels. The supramolecular network capable to swelling is created by cross-linking of the biopolymers using various reagents. Many cross-linking agents has been tested for last years, however, researchers still are looking for a new, more secure reactants. Squaric acid, 3,4-dihydroxy 3-cyclobutene 1,2- dione, is a very strong acid, which possess flat and rigid structure. Due to the presence of two carboxyl groups the squaric acid willingly reacts with amino groups of collagen. The main purpose of this study was to investigate the influence of addition of squaric acid on the chemical, physical and biological properties of collagen materials. The collagen type I was extracted from rat tail tendons and 1% solution in 0.1M acetic acid was prepared. The samples were cross-linked by the addition of 5%, 10% and 20% of squaric acid. The mixtures of all reagents were incubated 30 min on magnetic stirrer and then dialyzed against deionized water. The FTIR spectra show that the collagen structure is not changed by cross-linking by squaric acid. Although the mechanical properties of the collagen material deteriorate, the temperature of thermal denaturation of collagen increases after cross-linking, what indicates that the protein network was created. The lyophilized collagen gels exhibit porous structure and the pore size decreases with the higher addition of squaric acid. Also the swelling ability is lower after the cross-linking. The in vitro study demonstrates that the materials are attractive for 3T3 cells. The addition of squaric acid causes formation of cross-ling bonds in the collagen materials and the transparent, stiff hydrogels are obtained. The changes of physicochemical properties of the material are typical for cross-linking process, except mechanical properties – it requires further experiments. However, the results let us to conclude that squaric acid is a suitable cross-linker for protein materials for medicine and tissue engineering.

Keywords: collagen, squaric acid, cross-linking, hydrogel

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Authors: Yen-Hao Su, Wan-Chun Tang, Ya-Wen Cheng, Peik Sia, Chi-Chen Huang, Yi-Chao Lee, Hsin-Yi Jiang, Ming-Heng Wu, I-Lu Lai, Jun-Wei Lee, Kuen-Haur Lee

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There is a wide range of drugs and combinations under investigation and/or approved over the last decade to treat colorectal cancer (CRC), but the 5-year survival rate remains poor at stages II–IV. Therefore, new, more efficient drugs still need to be developed that will hopefully be included in first-line therapy or overcome resistance when it appears, as part of second- or third-line treatments in the near future. In this study, we revealed that heat shock protein 90 (Hsp90) inhibitors have high therapeutic potential in CRC according to combinative analysis of NCBI's Gene Expression Omnibus (GEO) repository and chemical genomic database of Connectivity Map (CMap). We found that second generation Hsp90 inhibitor, NVP-AUY922, significantly down regulated the activities of a broad spectrum of kinases involved in regulating cell growth arrest and death of NVPAUY922-sensitive CRC cells. To overcome NVP-AUY922-induced upregulation of survivin expression which causes drug insensitivity, we found that combining berberine (BBR), a herbal medicine with potency in inhibiting survivin expression, with NVP-AUY922 resulted in synergistic antiproliferative effects for NVP-AUY922-sensitive and -insensitive CRC cells. Furthermore, we demonstrated that treatment of NVP-AUY922-insensitive CRC cells with the combination of NVP-AUY922 and BBR caused cell growth arrest through inhibiting CDK4 expression and induction of microRNA-296-5p (miR-296-5p)-mediated suppression of Pin1–β-catenin–cyclin D1 signaling pathway. Finally, we found that the expression level of Hsp90 in tumor tissues of CRC was positively correlated with CDK4 and Pin1 expression levels. Taken together, these results indicate that combination of NVP-AUY922 and BBR therapy can inhibit multiple oncogenic signaling pathways of CRC.

Keywords: berberine, colorectal cancer, connectivity map, heat shock protein 90 inhibitor

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Authors: Xiaoyang Zhao, Kaiying Wang

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The choice of stocking density in poultry farming is a potential way for determining welfare level of poultry. However, it is difficult to measure stocking densities in poultry farming because of a lot of variables such as species, age and weight, feeding way, house structure and geographical location in different broiler houses. A method was proposed in this paper to measure the differences of young broilers reared under different stocking densities by sound analysis. Vocalisations of broilers were recorded and analysed under different stocking densities to identify the relationship between sounds and stocking densities. Recordings were made continuously for three-week-old chickens in order to evaluate the variation of sounds emitted by the animals at the beginning. The experimental trial was carried out in an indoor reared broiler farm; the audio recording procedures lasted for 5 days. Broilers were divided into 5 groups, stocking density treatments were 8/m², 10/m², 12/m² (96birds/pen), 14/m² and 16/m², all conditions including ventilation and feed conditions were kept same except from stocking densities in every group. The recordings and analysis of sounds of chickens were made noninvasively. Sound recordings were manually analysed and labelled using sound analysis software: GoldWave Digital Audio Editor. After sound acquisition process, the Mel Frequency Cepstrum Coefficients (MFCC) was extracted from sound data, and the Support Vector Machine (SVM) was used as an early detector and classifier. This preliminary study, conducted in an indoor reared broiler farm shows that this method can be used to classify sounds of chickens under different densities economically (only a cheap microphone and recorder can be used), the classification accuracy is 85.7%. This method can predict the optimum stocking density of broilers with the complement of animal welfare indicators, animal productive indicators and so on.

Keywords: broiler, stocking density, poultry farming, sound monitoring, Mel Frequency Cepstrum Coefficients (MFCC), Support Vector Machine (SVM)

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Authors: Kalep Filli, Sophia Wassén, Annika Krona, Mats Stading

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Food security challenge and the growing population in Sub-Saharan Africa centers on its agricultural transformation, where about 70% of its population is directly involved in farming. Research input can create economic opportunities, reduce malnutrition and poverty, and generate faster, fairer growth. Africa is discarding $4 billion worth of grain annually due to pre and post-harvest losses. Grains and tubers play a central role in food supply in the region but their production has generally lagged behind because no robust scientific input to meet up with the challenge. The African grains are still chronically underutilized to the detriment of the well-being of the people of Africa and elsewhere. The major reason for their underutilization is because they are under-researched. Any commitment by scientific community to intervene needs creative solutions focused on innovative approaches that will meet the economic growth. In order to mitigate this hurdle, co-creation activities and initiatives are necessary.An example of such initiatives has been initiated through Modibbo Adama University of Technology Yola, Nigeria and RISE (The Research Institutes of Sweden) Gothenburg, Sweden. Exchange of expertise in research activities as a possibility to create channel for value addition to agricultural commodities in the region under the ´Traditional Grain Network programme´ is in place. Process technologies, such as extrusion offers the possibility of creating products in the food and feed sectors, with better storage stability, added value, lower transportation cost and new markets. The Swedish–Nigerian initiative has focused on the development of high protein pasta. Dry microscopy of pasta sample result shows a continuous structural framework of proteins and starch matrix. The water absorption index (WAI) results showed that water was absorbed steadily and followed the master curve pattern. The WAI values ranged between 250 – 300%. In all aspect, the water absorption history was within a narrow range for all the eight samples. The total cooking time for all the eight samples in our study ranged between 5 – 6 minutes with their respective dry sample diameter ranging between 1.26 – 1.35 mm. The percentage water solubility index (WSI) ranged from 6.03 – 6.50% which was within a narrow range and the cooking loss which is a measure of WSI is considered as one of the main parameters taken into consideration during the assessment of pasta quality. The protein contents of the samples ranged between 17.33 – 18.60 %. The value of the cooked pasta firmness ranged from 0.28 - 0.86 N. The result shows that increase in ratio of cowpea flour and level of pregelatinized cowpea tends to increase the firmness of the pasta. The breaking strength represent index of toughness of the dry pasta ranged and it ranged from 12.9 - 16.5 MPa.

Keywords: cowpea, extrusion, gluten free, high protein, pasta, sorghum

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