Search results for: fibrinogen like protein 2

Authors: K. Thiel, P. Patrikainen, C. Nagy, D. Fitzpatrick, E.-M. Aro, P. Kallio

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Photosynthetic cyanobacteria have been recognized as potential future biotechnological hosts for the direct conversion of CO₂ into chemicals of interest using sunlight as the solar energy source. However, in order to develop commercially viable systems, the flux of electrons from the photosynthetic light reactions towards specified target chemicals must be significantly improved. The objective of the study was to investigate whether the autotrophic production efficiency of specified end-metabolites can be improved in engineered cyanobacterial cells by rescuing excited electrons that are normally lost to molecular oxygen due to the cyanobacterial flavodiiron protein Flv1/3. Natively Flv1/3 dissipates excess electrons in the photosynthetic electron transfer chain by directing them to molecular oxygen in Mehler-like reaction to protect photosystem I. To evaluate the effect of flavodiiron inactivation on autotrophic production efficiency in the cyanobacterial host Synechocystis sp. PCC 6803 (Synechocystis), sucrose was selected as the quantitative reporter and a representative of a potential end-product of interest. The concept is based on the native property of Synechocystis to produce sucrose as an intracellular osmoprotectant when exposed to high external ion concentrations, in combination with the introduction of a heterologous sucrose permease (CscB from Escherichia coli), which transports the sucrose out from the cell. In addition, cell growth, photosynthetic gas fluxes using membrane inlet mass spectrometry and endogenous storage compounds were analysed to illustrate the consequent effects of flv deletion on pathway flux distributions. The results indicate that a significant proportion of the electrons can be lost to molecular oxygen via Flv1/3 even when the cells are grown under high CO₂ and that the inactivation of flavodiiron activity can enhance the photosynthetic electron flux towards optionally available sinks. The flux distribution is dependent on the light conditions and the genetic context of the Δflv mutants, and favors the production of either sucrose or one of the two storage compounds, glycogen or polyhydroxybutyrate. As a conclusion, elimination of the native Flv1/3 reaction and concomitant introduction of an engineered product pathway as an alternative sink for excited electrons could enhance the photosynthetic electron flux towards the target endproduct without compromising the fitness of the host.

Keywords: cyanobacterial engineering, flavodiiron proteins, redirecting electron flux, sucrose

Procedia PDF Downloads 111

Authors: Elvan G. Bulut, Betul Goksun, Tugba G. Gun, Ozge Sakiyan Demirkol, Kamuran Ayhan, Kezban Candogan

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Dysphagia, difficulty in swallowing solid foods and thin liquids, is one of the common health threats among the elderly who require foods with modified texture in their diet. Although there are some commercial food formulations or hydrocolloids to thicken the liquid foods for dysphagic individuals, there is still a need for developing and offering new food products with enriched nutritional, textural and sensory characteristics to safely nourish these patients. 3D food printing is an appealing alternative in creating personalized foods for this purpose with attractive shape, soft and homogenous texture. In order to modify texture and prevent phase separation, hydrocolloids are generally used. In our laboratory, an optimized 3D printed beef-based formulation specifically for people with swallowing difficulties was developed based on the research project supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK Project # 218O017). The optimized formulation obtained from response surface methodology was 60% beef powder, 5.88% gelatin, and 0.74% kappa-carrageenan (all in a dry basis). This product was enriched with powders of freeze-dried beet, celery, and red capia pepper, butter, and whole milk. Proximate composition (moisture, fat, protein, and ash contents), pH value, CIE lightness (L*), redness (a*) and yellowness (b*), and color difference (ΔE*) values were determined. Counts of total mesophilic aerobic bacteria (TMAB), lactic acid bacteria (LAB), mold and yeast, total coliforms were conducted, and detection of coagulase positive S. aureus, E. coli, and Salmonella spp. were performed. The 3D printed products had 60.11% moisture, 16.51% fat, 13.68% protein, and 1.65% ash, and the pH value was 6.19, whereas the ΔE* value was 3.04. Counts of TMAB, LAB, mold and yeast and total coliforms before and after 3D printing were 5.23-5.41 log cfu/g, < 1 log cfu/g, < 1 log cfu/g, 2.39-2.15 log EMS/g, respectively. Coagulase positive S. aureus, E. coli, and Salmonella spp. were not detected in the products. The data obtained from this study based on determining some important product characteristics of functional beef-based formulation provides an encouraging basis for future research on the subject and should be useful in designing mass production of 3D printed products of similar composition.

Keywords: beef, dysphagia, product characteristics, texture-modified foods, 3D food printing

Procedia PDF Downloads 98

Authors: Alice Nwanyioma Ohuoba, Rose Erdoo Kukwa, Ukpabi Joseph Ukpabi

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Cassava root (Manihot esculenta) is one of the important carbohydrates containing crops in Nigeria. It is a staple food, mostly in the southern part of the country, and a source of income to farmers and processors. Cassava gari processing methods result to residue fiber (solid waste) from the sieving operation, these residue fibers ( solid wastes) can be dried and milled into flour and used to prepare cakes, cookies, crackers and chin-chin instead of being thrown away mostly on farmland or near the residential area. Flour for baking or frying may contain carbohydrates and protein (wheat flour) or rich in only carbohydrates (cassava flour). Cake, cookies, crackers, and chin-chin were prepared using the residue flour obtained from the residue fiber of cassava variety NR87184 roots, processed into gari. This study is aimed at evaluating the proximate composition, mineral content and sensory attributes of these selected snacks produced. The proximate composition results obtained showed that crackers had the lowest value in moisture (2.3390%) and fat (1.7130%), but highest in carbohydrates (85.2310%). Amongst the food products, cakes recorded the highest value in protein (8.0910%). Crude fibre values ranges from 2.5265% (cookies) to 3.4165% (crackers). The result of the mineral contents showed cookies ranking the highest in Phosphorus (65.8535 ppm) and Iron (0.1150 mg/L), Calcium (1.3800mg/L) and Potassium (7.2850 mg/L) contents, while chin-chin and crackers were lowest in Sodium ( 2.7000 mg/L). The food products were also subjected to sensory attributes evaluation by thirty member panelists using 9-hedonic scale which ranged from 1 ( dislike extremely) to 9 (like extremely). The means score obtained shows all the food products having above 7.00 (above “like moderately”). This study has shown that food products that may be functional or nutraceuticals could be prepared from the residue flour. There is a call for the use of gluten-free flour in baking due to ciliac disease and other allergic causes by gluten. Therefore local carbohydrates food crops like cassava residue flour that are gluten-free, could be the solution. In addition, this could aid cassava gari processing waste management thereby reducing post-harvest losses of cassava root.

Keywords: allergy, flour, food-products, gluten-free

Procedia PDF Downloads 139

Authors: Valencia Fernandes, Dharmendra Kumar Khatri, Shashi Bala Singh

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Background and Aim: Epigenetics are the inaudible signatures of several pathological processes in the brain. This study understands the influence of DNA methylation, a major epigenetic modification, in the prefrontal cortex and hippocampus of the diabetic brain and its notable effect on the cellular chaperones and synaptic proteins. Method: Chronic high fat diet and STZ-induced diabetic mice were studied for cognitive dysfunction, and global DNA methylation, as well as DNA methyltransferase (DNMT) activity, were assessed. Further, the cellular chaperones and synaptic proteins were examined using DNMT inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC)-via intracerebroventricular injection. Moreover, % methylation of these synaptic proteins were also studied so as to correlate its epigenetic involvement. Computationally, its interaction with the DNMT enzyme were also studied using bioinformatic tools. Histological studies for morphological alterations and neuronal degeneration were also studied. Neurogenesis, a characteristic marker for new learning and memory formation, was also assessed via the BrdU staining. Finally, the most important behavioral studies, including the Morris water maze, Y maze, passive avoidance, and Novel object recognition test, were performed to study its cognitive functions. Results: Altered global DNA methylation and increased levels of DNMTs within the nucleus were confirmed in the cortex and hippocampus of the diseased mice, suggesting hypermethylation at a genetic level. Treatment with AzadC, a global DNA demethylating agent, ameliorated the protein and gene expression of the cellular chaperones and synaptic fidelity. Furthermore, the methylation analysis profile showed hypermethylation of the hsf1 protein, a master regulator for chaperones and thus, confirmed the epigenetic involvement in the diseased brain. Morphological improvements and decreased neurodegeneration, along with enhanced neurogenesis in the treatment group, suggest that epigenetic modulations do participate in learning and memory. This is supported by the improved behavioral test battery seen in the treatment group. Conclusion: DNA methylation could possibly accord in dysregulating the memory-associated proteins at chronic stages in type 2 diabetes. This could suggest a substantial contribution to the underlying pathophysiology of several metabolic syndromes like insulin resistance, obesity and also participate in transitioning this damage centrally, such as cognitive dysfunction.

Keywords: epigenetics, cognition, chaperones, DNA methylation

Procedia PDF Downloads 186

Authors: Nahush Modak, Meena Pangarkar, Anand Pathak, Ankita Tamhane

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Background: Breast cancer is the prominent cause of cancer and mortality among women. This study was done to show the statistical analysis of a cohort of over 250 patients detected with breast cancer diagnosed by oncologists using Immunohistochemistry (IHC). IHC was performed by using ER; PR; HER2; Ki-67 antibodies. Materials and methods: Formalin fixed Paraffin embedded tissue samples were obtained by surgical manner and standard protocol was followed for fixation, grossing, tissue processing, embedding, cutting and IHC. The Ventana Benchmark XT machine was used for automated IHC of the samples. Antibodies used were supplied by F. Hoffmann-La Roche Ltd. Statistical analysis was performed by using SPSS for windows. Statistical tests performed were chi-squared test and Correlation tests with p<.01. The raw data was collected and provided by National Cancer Insitute, Jamtha, India. Result: Luminal B was the most prevailing molecular subtype of Breast cancer at our institute. Chi squared test of homogeneity was performed to find equality in distribution and Luminal B was the most prevalent molecular subtype. The worse prognostic indicator for breast cancer depends upon expression of Ki-67 and her2 protein in cancerous cells. Our study was done at p <.01 and significant dependence was observed. There exists no dependence of age on molecular subtype of breast cancer. Similarly, age is an independent variable while considering Ki-67 expression. Chi square test performed on Human epidermal growth factor receptor 2 (HER2) statuses of patients and strong dependence was observed in percentage of Ki-67 expression and Her2 (+/-) character which shows that, value of Ki depends upon Her2 expression in cancerous cells (p<.01). Surprisingly, dependence was observed in case of Ki-67 and Pr, at p <.01. This shows that Progesterone receptor proteins (PR) are over-expressed when there is an elevation in expression of Ki-67 protein. Conclusion: We conclude from that Luminal B is the most prevalent molecular subtype at National Cancer Institute, Jamtha, India. There was found no significant correlation between age and Ki-67 expression in any molecular subtype. And no dependence or correlation exists between patients’ age and molecular subtype. We also found that, when the diagnosis is Luminal A, out of the cohort of 257 patients, no patient shows >14% Ki-67 value. Statistically, extremely significant values were observed for dependence of PR+Her2- and PR-Her2+ scores on Ki-67 expression. (p<.01). Her2 is an important prognostic factor in breast cancer. Chi squared test for Her2 and Ki-67 shows that the expression of Ki depends upon Her2 statuses. Moreover, Ki-67 cannot be used as a standalone prognostic factor for determining breast cancer.

Keywords: breast cancer molecular subtypes , correlation, immunohistochemistry, Ki-67 and HR, statistical analysis

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Authors: Faozia A. S. Abuhtana, Yahia S. Abujnah, Said O. Gnann

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Ultra High Temperature (UHT) processed milk is widely distributed and preferred in numerous countries all over the world due its relatively high quality and long shelf life. Because of the notable high consumption rate of UHT in Libya in addition to negligible studies related to such product on the local level, this study was designed to assess the shelf life of locally produced as well as imported reconstituted sterilized whole milk samples marketed in Tripoli, Libya . Four locally produced vs. three imported brands were used in this study. All samples were stored at room temperature (25± 2C ) for 8 month long period, and subjected to physical, chemical, microbiological and sensory tests. These tests included : measurement of pH, specific gravity, percent acidity, and determination of fat, protein and melamine content. Microbiological tests included total aerobic count, total psychotropic bacteria, total spore forming bacteria and total coliform counts. Results indicated no detection of microbial growth of any type during the study period, in addition to no detection of melamine in all samples. On the other hand, a gradual decline in pH accompanied with gradual increase in % acidity of both locally produced and imported samples was observed. Such changes in both pH and % acidity reached their lowest and highest values respectively during the 24th week of storage. For instance pH values were (6.40, 6.55, 6.55, 6.15) and (6.30, 6.50, 6.20) for local and imported brands respectively. On the other hand, % acidity reached (0.185, 0181, 0170, 0183) and (0180, 0.180, 0.171) at the 24th week for local and imported brands respectively. Similar pattern of decline was also observed in specific gravity, fat and protein content in some local and imported samples especially at later stages of the study. In both cases, some of the recorded pH values, % acidity, sp. gravity and fat content were in violation of the accepted limits set by Libyan standard no. 356 for sterilized milk. Such changes in pH, % acidity and other UHT sterilized milk constituents during storage were coincided with a gradual decrease in the degree of acceptance of the stored milk samples of both types as shown by sensory scores recorded by the panelists. In either case degree of acceptance was significantly low at late stages of storage and most milk samples became relatively unacceptable after the 18th and 20th week for both untrained and trained panelists respectively.

Keywords: UHT milk, shelf life, quality, gravity, bacteria

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Authors: Supatchalee Sirichokworrakit, Pannin Intasen, Chansuda Angkawut

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Hom Nin rice (Oryza Sativa L.) was processed into flour and used to substitute wheat flour in donuts. The donuts were prepared with 0, 20, 40, 60, and 80% Hom Nin rice flour (HNF). The donuts were subjected to proximate, texture, color and sensory evaluations. The results of the study revealed that the ash, moisture, crude fiber contents increased while crude fat and protein contents decreased as the level of HNF increased. The hardness and chewiness of donut increased as the HNF increased but the cohesiveness, springiness, and specific volume decreased. Color of donut (L*, a*, and b* values) decreased with the addition of HNF. Overall acceptability for the 20-40% HNF additions did not differ significantly from the score of the 100% wheat flour.

Keywords: Hom Nin rice, donut, texture evaluation, sensory evaluation

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Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy

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Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.

Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract

Procedia PDF Downloads 317

Authors: Young June Yoon

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We will present four different topics in estimating the mechanical properties of biological tissues. First we elucidate the viscoelastic behavior of collagen molecules whose diameter is a couple of nanometers. By using the molecular dynamics simulation, we observed the viscoelastic behavior in different pulling velocity. Second, the protein layer, so called ‘sheath’ in enamel microstructure reduces the stress concentration in enamel minerals. We examined the result by using the finite element methods. Third, the anisotropic elastic constants of dentin are estimated by micromechanical analysis and estimated results are close to the experimentally measured data. Last, new formulation between the fabric tensor and the wave velocity is established for calcaneus by employing the poroelasticity. This formulation can be simply used for future experiments.

Keywords: tissues, mechanics, mechanical properties, wave propagation

Procedia PDF Downloads 355

Authors: Kuate Tueguem William Norbert, Ngoh Dooh Jules Patrice, Kone Sangou Abdou Nourou, Mboussi Serge Bertrand, Chewachang Godwill Mih, Essome Sale Charles, Djuissi Tohoto Doriane, Ambang Zachee

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The objectives of this study were to evaluate the effects of foliar application of 24-epibrassinolide (EBR) on the development of rice helminthosporiosis caused by Bipolaris oryzae and its influence on the improvement of growth parameters and induction of the synthesis of defense substances in the rice plants. The experimental asset up involved a multifactorial split-plot with two varieties (NERICA 3 and local variety KAMKOU) and five treatments (T0: control, T1: EBR, T2: BANKO PLUS (fungicide), T3: NPK (chemical fertilizer), T4: mixture: NPK + BANKO PLUS + EBR) with three repetitions. Agro-morphological and epidemiological parameters, as well as substances for plant resistance, were evaluated over two growing seasons. The application of the EBR induced significant growth of the rice plants for the 2015 and 2016 growing seasons on the two varieties tested compared to the T0 treatment. At 74 days after sowing (DAS), NERICA 3 showed plant heights of 58.9 ± 5.4; 83.1 ± 10.4; 86.01 ± 9.4; 69.4 ± 11.1 and 87.12 ± 7.4 cm at T0; T1; T2; T3, and T4, respectively. Plant height for the variety KAMKOU varied from 87,12 ± 8,1; 88.1 ± 8.1 and 92.02 ± 6.3 cm in T1, T2, and T3 to 74.1 ± 8.6 and 74.21 ± 11.4 cm in T0 and T3. In accordance with the low rate of expansion of helminthosporiosis in experimental plots, EBR (T1) significantly reduced the development of the disease with severities of 0.0; 1.29, and 2.04%, respectively at 78; 92, and 111 DAS on the variety NERICA 3 compared with1; 3.15 and 3.79% in the control T0. The reduction of disease development/severity as a result of the application of EBR is due to the induction of acquired resistance of rice varieties through increased phenol (13.73 eqAG/mg/PMF) and total protein (117.89 eqBSA/mg/PMF) in the T1 treatment against 5.37 eqAG/mg/PMF and 104.97 eqBSA/mg/PMF in T0 for the NERICA 3 variety. Similarly, on the KAMKOU variety, 148.53 eqBSA/mg/PMF were protein and 6.10 eqAG/mg/PMF of phenol in T1. In summary, the results show the significant effect of EBR on plant growth, yield, synthesis of secondary metabolites and defense proteins, and disease resistance. The EBR significantly reduced losses of rice grains by causing an average gain of about 1.55 t/ha compared to the control and 1.00 t/ha compared to the NPK-based treatment for the two varieties studied. Further, the enzymatic activities of PPOs, POXs, and PR2s were higher in leaves from treated EBR-based plants. These results show that 24-epibrassinolide can be used in the control of helminthosporiosis of rice to reduce disease and increase yields.

Keywords: Oryza sativa, 24-epibrassinolide, helminthosporiosis, secondary metabolites, PR proteins, acquired resistance

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Authors: Elizalde D. Bana

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Healing restores continuity and form through cell replication; hence, conserving structural integrity. In response to the worldwide pressing problem of chronic wounds in the healthcare delivery system, the researcher aims to provide effective intervention to preserve the structural integrity of the person. The wound healing effects of cryopreserved and lyophilized amniotic membrane (AM) of a term fetus embedded into two (2) concentrations (1.5 % and 1.0 %) of absorption-based ointment has been evaluated in vivo using the excision wound healing model 1x1 cm size. The total protein concentration in full term fetus was determined by the Biuret and Bradford methods, which are based on UV-visible spectroscopy. The percentages of protein presence in 9.5 mg (Mass total sample) of Amniotic membrane ranges between 14.77 – 14.46 % in Bradford method, while slightly lower to 13.78 – 13.80 % concentration in Biuret method, respectively. Bradford method evidently showed higher sensitivity for proteins than Biuret test. Overall, the amniotic membrane is composed principally of proteins in which a copious amount of literature substantially proved its healing abilities. After which, an area of 1 cm by 1 cm skin tissue was excised to its full thickness from the dorsolateral aspect of the isogenic mice and was applied twice a day with the ointment formulation having two (2) concentrations for the diabetic group and non-diabetic group. The wounds of each animal were left undressed and its area was measured every other day by a standard measurement formula from day 2,4,6,8,10,12 and 14. By the 14th day, the ointment containing 1.5 % of AM in absorption-based ointment applied to non-diabetic and diabetic group showed 100 % healing. The wound areas in the animals treated with the standard antibiotic, Mupirocin Ointment (Brand X) showed a 100% healing by the 14th day but with traces of scars, indicating that AM prepared from cryopreservation and lyophilization, at that given concentration, had a better wound healing property than the standard antibiotic. Four (4) multivariate tests were used which showed a significant interaction between days and treatments, meaning that the ointments prepared in two differing concentrations and induced in different groups of the mice had a significant effect on the percent of contraction over time. Furthermore, the evaluations of its effectiveness to wound healing were all significant although in differing degrees. It is observed that the higher the concentrations of amniotic membrane, the more effective are the results.

Keywords: wounds, healing, amniotic membrane ointments, biomedical, stem cell

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Authors: Ikhlas Ahmed, Jamillah Zamoon

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The human receptor for advanced glycation end product is a plasma membrane receptor with an intrinsically disordered region. The protein consists of three extracellular domains, a single membrane spanning transmembrane domain, and a cytosolic domain which is intrinsically disordered and responsible for signaling. The disordered nature of the cytosolic domain allows it to be dynamic in solution. This receptor self-associates to higher forms. The association is triggered by ligand, metal or by the extracellular domain. Fluorescence spectroscopy technique is used to test the self-association of the different concentrations of the cytosolic domain. This work has concluded that the cytosolic domain of this receptor also self-associates. Moreover, the self-association does not require ligand or metal.

Keywords: fluorescence spectroscopy, hRAGE, IDP, Self-association

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Authors: Jeongyeob Hong, Myeonghoon Park, Taeson Yoon

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After the advent of Achaeans which utilize different metabolism pathway and contain conspicuously different cellular structure, they have been recognized as possible materials for developing quality of human beings. Among diverse Achaeans, in this paper, we compared 16s RNA Sequences of four different species of Thermococcus: Achaeans genus specialized in sulfur-dealing metabolism. Four Species, Barophilus, Kodakarensis, Hydrothermalis, and Onnurineus, live near the hydrothermal vent that emits extreme amount of sulfur and heat. By comparing ribosomal sequences of aforementioned four species, we found similarities in their sequences and expressed protein, enabling us to expect that certain ribosomal sequence or proteins are vital for their survival. Apriori algorithms and Decision Tree were used. for comparison.

Keywords: Achaeans, Thermococcus, apriori algorithm, decision tree

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Authors: M. Ekiert, T. Uhl, A. Mlyniec

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Tendons are the biological soft tissue structures composed of collagen, proteoglycan, glycoproteins, water and cells of extracellular matrix (ECM). Tendons, which primary function is to transfer force generated by the muscles to the bones causing joints movement, are exposed to many micro and macro damages. In fact, tendons and ligaments trauma are one of the most numerous injuries of human musculoskeletal system, causing for many people (particularly for athletes and physically active people), recurring disorders, chronic pain or even inability of movement. The number of tendons reconstruction and transplantation procedures is increasing every year. Therefore, studies on soft tissues storage conditions (influencing i.e. tissue aging) seem to be an extremely important issue. In this study, an atomic-scale investigation on the kinetics of decomposition of two selected tendon components – collagen type I (which forms a 60-85% of a tendon dry mass) and elastin protein (which combine with ECM creates elastic fibers of connective tissues) is presented. A molecular model of collagen and elastin was developed based on crystal structure of triple-helical collagen-like 1QSU peptide and P15502 human elastin protein, respectively. Each model employed 4 linear strands collagen/elastin strands per unit cell, distributed in 2x2 matrix arrangement, placed in simulation box filled with water molecules. A decomposition phenomena was simulated with molecular dynamics (MD) method using ReaxFF force field and periodic boundary conditions. A set of NVT-MD runs was performed for 1000K temperature range in order to obtained temperature-depended rate of production of decomposition by-products. Based on calculated reaction rates activation energies and pre-exponential factors, required to formulate Arrhenius equations describing kinetics of decomposition of tested soft tissue components, were calculated. Moreover, by adjusting a model developed for collagen, system scalability and correct implementation of the periodic boundary conditions were evaluated. An obtained results provide a deeper insight into decomposition of selected tendon components. A developed methodology may also be easily transferred to other connective tissue elements and therefore might be used for further studies on soft tissues aging.

Keywords: decomposition, molecular dynamics, soft tissue, tendons

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Authors: Yuanyuan Zhang, Yujuan Zheng, Giuseppina Barutello, Sumako Kameishi, Kungchun Chiu, Katharina Hennig, Martial Balland, Federica Cavallo, Lars Holmgren

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Angiogenesis describes that new blood vessels migrate from pre-existing ones to form 3D lumenized structure and remodeling. During directional migration toward the gradient of pro-angiogenic factors, the endothelial cells, especially the tip cells need filopodia to sense the environment and exert the pulling force. Of particular interest are the integrin proteins, which play an essential role in focal adhesion in the connection between migrating cells and extracellular matrix (ECM). Understanding how these biomechanical complexes orchestrate intrinsic and extrinsic forces is important for our understanding of the underlying mechanisms driving angiogenesis. We have previously identified Angiomotin (Amot), a member of Amot scaffold protein family, as a promoter for endothelial cell migration in vitro and zebrafish models. Hence, we established inducible endothelial-specific Amot knock-out mice to study normal retinal angiogenesis as well as tumor angiogenesis. We found that the migration ratio of the blood vessel network to the edge was significantly decreased in Amotec- retinas at postnatal day 6 (P6). While almost all the Amot defect tip cells lost migration advantages at P7. In consistence with the dramatic morphology defect of tip cells, there was a non-autonomous defect in astrocytes, as well as the disorganized fibronectin expression pattern correspondingly in migration front. Furthermore, the growth of transplanted LLC tumor was inhibited in Amot knockout mice due to fewer vasculature involved. By using MMTV-PyMT transgenic mouse model, there was a significantly longer period before tumors arised when Amot was specifically knocked out in blood vessels. In vitro evidence showed that Amot binded to beta-actin, Integrin beta 1 (ITGB1), Fibronectin, FAK, Vinculin, major focal adhesion molecules, and ITGB1 and stress fibers were distinctly induced by Amot transfection. Via traction force microscopy, the total energy (force indicater) was found significantly decreased in Amot knockdown cells. Taken together, we propose that Amot is a novel partner of the ITGB1/Fibronectin protein complex at focal adhesion and required for exerting force transition between endothelial cell and extracellular matrix.

Keywords: angiogenesis, angiomotin, endothelial cell migration, focal adhesion, integrin beta 1

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Authors: Misheck Musokwa, Paramu Mafongoya, Simon Lorentz

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Seasonal shortages of fodder during the dry season is a major constraint to smallholder livestock farmers in South Africa. To mitigate the shortage of fodder, legume trees can be intercropped with pastures which can diversify the sources of feed and increase the amount of protein for grazing animals. The objective was to evaluate dry matter yield of Panicum maximum and land productivity under different fodder production systems during 2016/17-2017/18 seasons at Empangeni (28.6391° S and 31.9400° E). A randomized complete block design, replicated three times was used, the treatments were sole Panicum maximum, Panicum maximum + Sesbania sesban, Panicum maximum + pigeonpea, sole Sesbania sesban, Sole pigeonpea. Three months S.sesbania seedlings were transplanted whilst pigeonpea was direct seeded at spacing of 1m x 1m. P. maximum seeds were drilled at a respective rate of 7.5 kg/ha having an inter-row spacing of 0.25 m apart. In between rows of trees P. maximum seeds were drilled. The dry matter yield harvesting times were separated by six months’ timeframe. A 0.25 m² quadrant randomly placed on 3 points on the plot was used as sampling area during harvesting P. maximum. There was significant difference P < 0.05 across 3 harvests and total dry matter. P. maximum had higher dry matter yield as compared to both intercrops at first harvest and total. The second and third harvest had no significant difference with pigeonpea intercrop. The results was in this order for all 3 harvest: P. maximum (541.2c, 1209.3b and 1557b) kg ha¹ ≥ P. maximum + pigeonpea (157.2b, 926.7b and 1129b) kg ha¹ > P. maximum + S. sesban (36.3a, 282a and 555a) kg ha¹. Total accumulation of dry matter yield of P. maximum (3307c kg ha¹) > P. maximum + pigeonpea (2212 kg ha¹) ≥ P. maximum + S. sesban (874 kg ha¹). There was a significant difference (P< 0.05) on seed yield for trees. Pigeonpea (1240.3 kg ha¹) ≥ Pigeonpea + P. maximum (862.7 kg ha¹) > S.sesbania (391.9 kg ha¹) ≥ S.sesbania + P. maximum. The Land Equivalent Ratio (LER) was in the following order P. maximum + pigeonpea (1.37) > P. maximum + S. sesban (0.84) > Pigeonpea (0.59) ≥ S. Sesbania (0.57) > P. maximum (0.26). Results indicates that it is beneficial to have P. maximum intercropped with pigeonpea because of higher land productivity. Planting grass with pigeonpea was more beneficial than S. sesban with grass or sole cropping in terms of saving the shortage of arable land. P. maximum + pigeonpea saves a substantial (37%) land which can be subsequently be used for other crop production. Pigeonpea is recommended as an intercrop with P. maximum due to its higher LER and combined production of livestock feed, human food, and firewood. Panicum grass is low in crude protein though high in carbohydrates, there is a need for intercropping it with legume trees. A farmer who buys concentrates can reduce costs by combining P. maximum with pigeonpea this will provide a balanced diet at low cost.

Keywords: fodder, livestock, productivity, smallholder farmers

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Authors: Mayuva Youngsabanant, Chanikarn Srinark, Supanyika Sengsai, Soratorn Kerdkriangkrai, Nongnuch Gumlungpat, Mayuree Pumipaiboon

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The follicular fluid proteins of healthy medium size follicles (4-6 mm in diameters) and large size follicles (7-8 mm in diameter) of large white pig ovaries were collected by using sterile technique. They were used for testing the effect on primary in vitro cell culture growth of porcine oviductal epithelial cells (pOEC). Porcine oviductal epithelial cells of luteal phase was culture in M199 and added with 10% fetal calf serum 2.2 mg/mL, NaHCO₃, 0.25 mM pyruvate, 15 µg/mL and 50 µg/mL, gentamycin sulfate at high humidified atmosphere with 5% CO₂ in 95% air atmosphere at 37°C for 96 h before testing. The optimized concentration of pFF of two follicle sizes (at concentration of 2, 4, 20, 40, 200, 400, 500, and 600 µg proteins) in culture medium was observed for 24 h using MTT assay. Results were analyzed with a one-way ANOVA in SPSS statistic. Moreover, pOEC was also studied in morphological characteristic on long-term culture. The results of long-term study revealed that pOEC showed 70-80 percentage of healthy morphology on epithelial-like character and contained 30 percentage of an elongated shape (fibroblast-like morphology) at 4 weeks of culture time. MTT assay reviewed an increase in the percentage of viability of pOEC in 2 treated of follicular fluid groups. Two treatment concentration groups were higher than control group (p < 0.05) but not in positive control group. Interestingly, at 200 µg protein of 2 treated follicular fluid groups were reached the highest cell viability which is higher than a positive control and it is significantly different form control group (P < 0.05). These cells are developed and had fibroblast elongate shape which is longer than the cells in control group and positive control group. This report implies that pFF of medium follicle size at 200 µg proteins and large follicle size at 200 and 500 µg proteins could be optimized concentration for using as a supplement in culture medium to promote cell growth and development instead of growth hormone from fetal calf serum. It could be applied in cell biotechnology researches. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: in vitro, porcine follicular fluid protein (pFF), porcine oviductal epithelial cells (pOEC), MTT

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Authors: Selvaprakash Ramalingam, Rabi N. Sahoo, Dharmendra Saraswat, A. Kumar, Rajeev Ranjan, Joydeep Mukerjee, Viswanathan Chinnasamy, K. K. Chaturvedi, Sanjeev Kumar

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Rice is one of the most important staple food crops in the world. Among the various diseases that affect rice crops, rice blast is particularly significant, causing crop yield and economic losses. While the plant has defense mechanisms in place, such as chemical indicators (proteins, salicylic acid, jasmonic acid, ethylene, and azelaic acid) and resistance genes in certain varieties that can protect against diseases, susceptible varieties remain vulnerable to these fungal diseases. Early prediction of rice blast (RB) disease is crucial, but conventional techniques for early prediction are time-consuming and labor-intensive. Hyperspectral remote sensing techniques hold the potential to predict RB disease at its asymptomatic stage. In this study, we aimed to demonstrate the prediction of RB disease at the asymptomatic stage using non-imaging hyperspectral ASD spectroradiometer under controlled laboratory conditions. We applied statistical spectral discrimination theory to identify unknown spectra of M. Oryzae, the fungus responsible for rice blast disease. The infrared (IR) region was found to be significantly affected by RB disease. These changes may result in alterations in the absorption, reflection, or emission of infrared radiation by the affected plant tissues. Our research revealed that the protein spectrum in the IR region is impacted by RB disease. In our study, we identified strong correlations in the region (Amide group - I) around X 1064 nm and Y 1300 nm with the Lambda / Lambda derived spectra methods for protein detection. During the stages when the disease is developing, typically from day 3 to day 5, the plant's defense mechanisms are not as effective. This is especially true for the PB-1 variety of rice, which is highly susceptible to rice blast disease. Consequently, the proteins in the plant are adversely affected during this critical time. The spectral contour plot reveals the highly correlated spectral regions 1064 nm and Y 1300 nm associated with RB disease infection. Based on these spectral sensitivities, we developed new spectral disease indices for predicting different stages of disease emergence. The goal of this research is to lay the foundation for future UAV and satellite-based studies aimed at long-term monitoring of RB disease.

Keywords: rice blast, asymptomatic stage, spectral sensitivity, IR

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Authors: Mojdeh Chelemal Dezfoul Nejad, Masomeh Moradi, Mehrzad Mesbah, Mehran Javaheri Babouli

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This study was conducted in order to characterize the different levels of dietary vitamin C on some of biochemical parameters of Barbuas grypus. For this purpose 300 Barbuas grypus were divided into 15 groups. five levels of vitamin C (0, 200 ,400,800,1600 mg kg-1 diet) and their combination were used to prepare five experimental diets. The fish were fed 3% of their wet b.wt. per day for a 60 days period. Blood samples were obtained from six fish of each tank at the end of experiment. The results reveal that fish fed diets containing 1600 mg kg^-1 vitamin C had the significant decreased in the mean amount of cholesterol, glucose and triglyceride (p<0.05). Also, there was no significant difference in the mean amount of total protein with the different diets designed for this experiment (p>0.05).

Keywords: Barbuas, grypus, vitamin C, biochemical parameters

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Authors: Somayeh Fani, Mahmood Ameen Abdulla

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Costus speciosus is an important Malaysian medicinal plant commonly used traditionally in the treatment of many aliments. The present investigation is designed to elucidate preventive effects of ethanolic extracts of C. speciosus rhizome against absolute ethanol-induced gastric mucosal injury in Sprague-Dawley rats. Five groups of rats were orally pre-treated with vehicle, carboxymethylcellulose (CMC) as normal control group (Group 1), ethanol as ulcer control group (Group 2), omeprazole 20 mg/kg (reference group) (Group 3), and 250 and 500 mg/kg of C. speciosus extract (experimental groups) (Group 4 and 5), respectively. An hour later, CMC was given orally to Group 1 rats and absolute ethanol was given orally to Group 2-5 rats to generate gastric mucosal injury. After an additional hour, the rats were sacrificed. Grossly, ulcer control group exhibited severe of gastric mucosal hemorrhagic injury and increased in ulcer area, whereas groups pre-treated with omeprazole or plant’s rhizomes exhibited the significant reduction of gastric mucosal injury. Significant increase in the pH and mucous of gastric content was observed in rats re-treated with C. speciosus rhizome. Histology, ulcer control rats, demonstrated remarkable disruption of gastric mucosa, increased in edema and inflammatory cells infiltration of submucosal layer compared to rats pre-treated with rhizomes extract. Periodic acid Schiff staining for glycoprotein, rats pre-fed with C. speciosus C. displayed remarkably intense uptake of magenta color by glandular gastric mucosa compared with ulcer control rats. Immunostaining of gastric epithelium, rats pre-treatment with rhizome extract provide evidence of up-regulation of HSP70 and down-regulation of Bax proteins compared to ulcer control animals. Gastric tissue homogenate, C. speciosus significantly increased the activity of superoxide dismutase (SOD), and catalase (CAT), increased the level of non-protein sulfhydryl (NP-SH) and decreased the level of lipid peroxidation after ethanol administration. Acute toxicity test did not show any signs of toxicity. The mechanisms implicated the gasrtoprotective property of C. speciosus depend upon the antisecretory activity, increased in gastric mucus glycoprotein, up-regulation of HSP70 protein and down-regulation of Bax proteins, reduction in the lipid peroxidation and increase in the level of NP-SH and antioxidant enzymes activity in gastic homogenate.

Keywords: antioxidant, Costus speciosus, gastric ulcer, histology, omeprazole

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Authors: Luz Stella Cadavid-Rodriguez, Viviana E. Castro-Lopez

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Fish waste (FW) has a high content of potentially biodegradable components, so it is amenable to be digested anaerobically. In this line, anaerobic digestion (AD) of FW has been studied for biogas production. Nevertheless, intermediate products such as volatile fatty acids (VFA), generated during the acidogenic stage, have been scarce investigated, even though they have a high potential as a renewable source of carbon. In the literature, there are few studies about the Inoculum-Substrate (I/S) ratio on acidogenesis. On the other hand, it is well known that pH is a critical factor in the production of VFA. The optimum pH for the production of VFA seems to change depending on the substrate and can vary in a range between 5.25 and 11. Nonetheless, the literature about VFA production from protein-rich waste, such as FW, is scarce. In this context, it is necessary to deepen on the determination of the optimal operating conditions of acidogenic fermentation for VFA production from protein-rich waste. Therefore, the aim of this research was to optimize the volatile fatty acid production from artisanal fishing waste, studying the effect of pH and the I/S ratio on the acidogenic process. For this research, the inoculum used was a methanogenic sludge (MS) obtained from a UASB reactor treating wastewater of a slaughterhouse plant, and the FW was collected in the port of Tumaco (Colombia) from the local artisanal fishers. The acidogenic fermentation experiments were conducted in batch mode, in 500 mL glass bottles as anaerobic reactors, equipped with rubber stoppers provided with a valve to release biogas. The effective volume used was 300 mL. The experiments were carried out for 15 days at a mesophilic temperature of 37± 2 °C and constant agitation of 200 rpm. The effect of 3 pH levels: 5, 7, 9, coupled with five I/S ratios, corresponding to 0.20, 0.15, 0.10, 0.05, 0.00 was evaluated taking as a response variable the production of VFA. A complete randomized block design was selected for the experiments in a 5x3 factorial arrangement, with two repetitions per treatment. At the beginning and during the process, pH in the experimental reactors was adjusted to the corresponding values of 5, 7, and 9 using 1M NaOH or 1M H2SO4, as was appropriated. In addition, once the optimum I/S ratio was determined, the process was evaluated at this condition without pH control. The results indicated that pH is the main factor in the production of VFA, obtaining the highest concentration with neutral pH. By reducing the I/S ratio, as low as 0.05, it was possible to maximize VFA production. Thus, the optimum conditions found were natural pH (6.6-7.7) and I/S ratio of 0.05, with which it was possible to reach a maximum total VFA concentration of 70.3 g Ac/L, whose major components were acetic acid (35%) and butyric acid (32%). The findings showed that the acidogenic fermentation of FW is an efficient way of producing VFA and that the operating conditions can be simple and economical.

Keywords: acidogenesis, artisanal fishing waste, inoculum to substrate ratio, volatile fatty acids

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Authors: K. V. Ramanaiah, R. Jagan, N. N. Murthy

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Trinuclear oxo-centered carboxylate-bridged iron complexes, [Fe3(µ3-O)(µ2-O2CR)L¬3]+/0 (where R = alkyl or aryl; L = H2O, ROH, Py, solvent) have attracted tremendous attention because of their interesting structural and magnetic properties, exhibit mixed-valent trapped and de-trapped states, and have bioinorganic relevance. The presence of a trinuclear iron binding center has been implicated in the formation of both bacterial and human iron storage protein, Ft. They are used as precursors for the synthesis of models for the active-site structures of non-heme proteins, hemerythrin (Hr), methane monooxygenase (MMO) and polyiron storage protein, ferritin (Ft). Used as important building blocks for the design and synthesis of supramolecules this can exhibit single molecular magnetism (SMM). Such studies have often employed simple and compact carboxylate ligands and the use of bulky carboxylates is scarce. In the present study, we employed two different type of sterically hindered carboxylates and synthesized a series of novel oxo-centered, carboxylate-bridged triiron complexes of general formula [Fe3(O)(O2CCPh3)6L3]X (L = H2O, 1; py, 2; 4-NMe2py, 3; X = ClO4; L = CH3CN, 4; X = FeCl4) and [Fe3(O)(O2C-anth)6L3]X (L = H2O, 5; X = ClO4; L = CH3OH, 6; X = Cl). Along with complex [Fe(OMe)2(O2CCPh3)]10, 7 was prepared by the self-assemble of anhydrous FeCl3, sodium triphenylacetate and sodium methoxide at ratio of 1:1:2 in CH3OH. The Electronic absorption spectra of these complexes 1-6, in CH2Cl2 display weak bands at near FTIR region (970-1135 nm, ε > 15M-1cm-1). For complex 7, one broad band centered at ~670nm and also an additional intense charge transfer (L→M or O→M) bands between 300 to 550nm observed for all the complexes. Paramagnetic 1H NMR is introduced as a good probe for the characterization of trinuclear oxo - cantered iron compounds in solution when the L ligand coordinated to iron varies as: H2O, py, 4-NMe2py, and CH3OH. The solution state magnetic moment values calculated by using Evans method for all the complexes and also solid state magnetic moment value of complex, 7 was calculated by VSM method, which is comparable with solution state value. These all magnetic moment values indicate there is a spin exchange process through oxo and carboxylate bridges in between two irons (d5). The ESI-mass data complement the data obtained from single crystal X-ray structure. Further purity of the compounds was confirmed by elemental analysis. Finally, structural determination of complexes 1, 3, 4, 5, 6 and 7 were unambiguously conformed by single crystal x-ray studies.

Keywords: decanuclear, paramagnetic NMR, trinuclear, uv-visible

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Authors: Claudia Cabezas-Zabala, Jairo Lindarte-Artunduaga, Carlos Mario Zuluaga-Dominguez

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Amaranth is an emerging pseudocereal rich in such essential nutrients as protein and dietary fiber, which was employed as an ingredient in the formulation of crackers to evaluate the rheological performance and sensory acceptability of the obtained food. A completely randomized factorial design was used with two factors: (A) ratio of wheat and amaranth flour used in the preparation of the dough, in proportion 90:10 and 80:20 (% w/w) and (B) two levels of inulin addition of 8.4% and 16.7 %, having two control doughs made from amaranth and wheat flour, respectively. Initially, the functional properties of the formulations mentioned were measured, showing no significant differences in the water absorption capacity (WAC) and swelling power (SP), having mean values between 1.66 and 1.81 g/g for WAC and between 1.75 and 1.86 g/g for SP, respectively. The amaranth flour had the highest water holding capacity (WHR) of 8.41 ± 0.15 g/g and emulsifying activity (EA) of 74.63 ± 1.89 g/g. Moreover, the rheological behavior, measured through the use of farinograph, extensograph, Mixolab, and falling index, showed that the formulation containing 20% of amaranth flour and 7.16% of inulin had a rheological behavior similar to the control produced exclusively with wheat flour, being the former, the one selected for the preparation of crackers. For this formulation, the farinograph showed a mixing tolerance index of 11 UB, indicating a strong and cohesive dough; likewise, the Mixolab showed dough reaches stability at 6.47 min, indicating a good resistance to mixing. On the other hand, the extensograph exhibited a dough resistance of 637 UB, as well as extensibility of 13.4 mm, which corresponds to a strong dough capable of resisting the laminate. Finally, the falling index was 318 s, which indicates the crumb will retain enough air to enhance the crispness of a characteristic cracker. Finally, a sensory consumer test did not show significant differences in the evaluation of aroma between the control and the selected formulation, while this latter had a significantly lower rating in flavor. However, a purchase intention of 70 % was observed among the population surveyed. The results obtained in this work give perspectives for the industrial use of amaranth in baked goods. Additionally, amaranth has been a product typically linked to indigenous populations in the Andean South American countries; therefore, the search for diversification and alternatives of use for this pseudocereal has an impact on the social and economic conditions of such communities. The technological versatility and nutritional quality of amaranth is an advantage for consumers, favoring the consumption of healthy products with important contributions of dietary fiber and protein.

Keywords: amaranth, crackers, rheology, pseudocereals, kneaded products

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Authors: Marcin Stasiulewicz, Sebastian Filipkowski, Aneta Panuszko

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Background: The hallmark of neurodegenerative diseases, including Alzheimer's and Parkinson's diseases, is an aggregation of the abnormal forms of peptides and proteins. Water is essential to functioning biomolecules, and it is one of the key factors influencing protein folding and misfolding. However, the hydration studies of proteins are complicated due to the complexity of protein systems. The use of model compounds can facilitate the interpretation of results involving larger systems. Objectives: The goal of the research was to characterize the properties of the hydration water surrounding the two three-residue K peptide fragments INS (Isoleucine - Asparagine - Serine) and NSR (Asparagine - Serine - Arginine). Methods: Fourier-transform infrared spectra of aqueous solutions of the tripeptides were recorded on Nicolet 8700 spectrometer (Thermo Electron Co.) Measurements were carried out at 25°C for varying molality of solute. To remove oscillation couplings from water spectra and, consequently, obtain narrow O-D semi-heavy water bands (HDO), the isotopic dilution method of HDO in H₂O was used. The difference spectra method allowed us to isolate the tripeptide-affected HDO spectrum. Results: The structural and energetic properties of water affected by the tripeptides were compared to the properties of pure water. The shift of the values of the gravity center of bands (related to the mean energy of water hydrogen bonds) towards lower values with respect to the ones corresponding to pure water suggests that the energy of hydrogen bonds between water molecules surrounding tripeptides is higher than in pure water. A comparison of the values of the mean oxygen-oxygen distances in water affected by tripeptides and pure water indicates that water-water hydrogen bonds are shorter in the presence of these tripeptides. The analysis of differences in oxygen-oxygen distance distributions between the tripeptide-affected water and pure water indicates that around the tripeptides, the contribution of water molecules with the mean energy of hydrogen bonds decreases, and simultaneously the contribution of strong hydrogen bonds increases. Conclusions: It was found that hydrogen bonds between water molecules in the hydration sphere of tripeptides are shorter and stronger than in pure water. It means that in the presence of the tested tripeptides, the structure of water is strengthened compared to pure water. Moreover, it has been shown that in the vicinity of the Asparagine - Serine - Arginine, water forms stronger and shorter hydrogen bonds. Acknowledgments: This work was funded by the National Science Centre, Poland (grant 2017/26/D/NZ1/00497).

Keywords: amyloids, K-peptide, hydration, FTIR spectroscopy

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Authors: Ž. Vaštag, Lj. Popović, S. Popović

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After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilized as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity

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Authors: Asma Bashir, Neha Farid, Kashif Ali, Kiran Fatima

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Background: Bacteriocins are a subclass of antimicrobial peptides that are becoming extremely important in treatments. It is possible to utilise bacteriocins in place of or in addition to traditional antibiotics. It is possible to treat a variety of infections, including Vancomycin-Resistant Staphylococcus aureus (VRSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), using the targeted spectrum of activity of these microorganisms. Method: This study aimed to examine the efficiency of antibiotics and bacteriocin against VRSA and MRSA. The effects of bacteriocins, such as enterocin KAE01, enterocin KAE03, enterocin KAE05, and enterocin KAE06 isolated from Enterococcus faecium strains, alone and in combination with vancomycin and methicillin antibiotics were examined. The selection technique utilized the minimum inhibitory concentrations (MICs) against Gram-positive indicator strain ATCC 6538 Methicillin-Resistant Staphylococcus aureus (MRSA) and indicator strain KSA 02 Vancomycin-Resistant Staphylococcus aureus (VRSA). Results: We report the isolation and identification of enterocins KAE01, KAE03, KAE05, and KAE06 from food isolates of Enterococcus faecium (KAE01, KAE03, KAE05, and KAE06). After isolating the protein, it was partially purified with ammonium sulphate precipitation and purified with fast protein liquid chromatography (FPLC) procedures. Combinations of enterocin KAE01, 1 citric acid, 1 lactic acid, and microcin J25, 1 reuterin, 1 citric acid, and microcin J25, 1 reuterin, 1 lactic acid shown synergistic benefits (FIC index = 0.5) against Vancomycin-Resistant Staphylococcus aureus (VRSA). In addition, a moderately synergistic (FIC index = 0.75) interaction was seen between pediocin PA-1, 1 citric acid, 1 lactic acid, and reuterin 1 citric acid, 1 lactic acid against L. ivanovii HPB28. In the presence of acids, nisin Z exhibited a modestly synergistic effect (FIC index = 0.625-0.75); however, it exhibited additive effects (FIC index = 1) when combined with reuterin or pediocin PA-1 against L. ivanovii HPB28. The efficacy of synergistic consortiums against Gram-positive bacteria was examined. Conclusion: Combining antimicrobials with various modes of action boosted efficacy and expanded the spectrum of inhibition, particularly against multidrug-resistant pathogens, according to our research.

Keywords: Enterococcus faecium, bacteriocin, antimicrobial resistance, antagonistic activity, vancomycin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus

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Authors: Hatice Akbıyık, Hülya Gökmen Özel, Nagehan Emiralioğlu, Elmas Ebru Güneş Yalçın, Deniz Doğru Ersöz, Hayriye Uğur Özçelik, Nural Kiper

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Background: Dietary recommendations in cystic fibrosis (CF) are based on the need to compensate for the increased energy needs of infection, the increased energy cost of breathing and the losses, incurred from malabsorption. Studies in CF indicate that dietary recommendations for CF patients can be difficult to achieve Aim: The aim of this study was to evaluate the energy and nutrient intakes and to compare in accordance with CF dietary guidelines in CF. Methods: One-hundred sixty patients with CF, aged between 2 to 20 years (mean±SD= 7.4±4.8 years) attending Hacettepe University, Faculty of Medicine, Department of Pediatric Pulmonary Diseases were included. Energy and nutrient intakes from foods and enteral products were calculated using a-24-hour dietary recall method with BEBIS 7.2 programme. Percentages of energy and nutrient intakes were compared in accordance with CF dietary guidelines. Patients or/and parents completed a questionnaire showing mealtime problems, usage of alternative therapies and type of nutrition. Statistical analyses were done using SPSS 16.0 programme. Results: It was obtained that 14.5% and 46.9% of the total energy intake were from proteins and carbohydrates, respectively. The actual contribution of total, saturated, monounsaturated and polyunsaturated fats to the total caloric intake was 37.5%, 14.3%, 14.9%, 9.9%, respectively. It was found that 87.7% of energy, 85% of protein 91.7% of carbohydrate, 81.1% of fat intakes were met, when compared CF recommended intakes of 120% RDA. Additionally 67%, 69.5%, 68.2% and 68.9% of the subjects did not achieve CF recommended intakes of 120% RDA for energy, protein, carbohydrate and fat, respectively. Patients with CF had low intakes for age for almost all vitamins and minerals, although supplementation was given. Especially most patients did not achieve the minimum recommended vitamin K intake of 120% RDA. The percentage meeting 120% RDA was 75.9% for vitamin K. It was shown that 41% of the patients had mealtime problems and they skipped the breakfast. Moreover 25.4% of the patients used alternative products outside the standard treatment (such as omega-3, ginger, turmeric, local honey). It was also showed that 60.8% of patients were using enteral products in addition to normal foods, the remaining patients were on only normal foods. Conclusion: The aims of improving nutritional status in children are to achieve normal weight gain and growth; optimize vitamin and mineral status; and slow the rate of clinical decline. In this study although enteral products were used in patients with CF, it was found that energy and nutrient requirements were unable to meet. Because dietary assessment is essential to identify the need for earlier nutritional intervention, in each visit patients need to be referred to CF specialist dietitian.

Keywords: cystic fibrosis, energy and nutrient intakes, mealtime problems, malabsorbtion

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Authors: Gaweł Magdalena, Lipkowska Anna, Olbert Magdalena, Frąckiewicz Ewelina, Librowski Tadeusz, Nowak Gabriel, Pilc Andrzej

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Heme oxygenase-1 (HO-1), also known as heat shock protein 32 (HSP32), has been shown to be implicated in cytoprotection in various organs. Its activation plays a significant role in acute and chronic inflammation, protecting cells from oxidative injury and apoptosis. This inducible isoform of HO catalyzes the first and rate-limiting step in heme degradation to produce equimolar quantities of biologically active products: carbon monoxide (CO), free iron and biliverdin. CO has been reported to possess anti-apoptotic properties. Moreover, it inhibits the production of proinflammatory cytokines and stimulates the synthesis of the anti-inflammatory interleukin-10 (IL-10), as well as promotes vasodilatation at sites of inflammation. The second product of catalytic HO-1 activity, free cytotoxic iron, is promptly sequestered into the iron storage protein ferritin, which lowers the pro-oxidant state of the cell. The third product, biliverdin, is subsequently converted by biliverdin reductase into the bile pigment bilirubin, the most potent endogenous antioxidant among the constituents of human serum, which modulates immune effector functions and suppresses inflammatory response. Furthermore, being one of the so-called stress proteins, HO-1 adaptively responds to different stressors, such as reactive oxygen species (ROS), inflammatory cytokines and heavy metals and thus protects cells against such conditions as ischemia, hemorrhagic shock, heat shock or hypoxia. It is suggested that pharmacologic modulation of HO-1 may represent an effective strategy for prevention of stress and drug-induced gastrointestinal toxicity. HO-1 is constitutively expressed in normal gastric, intestinal and colonic mucosa and up-regulated during inflammation. It has been proven that HO-1 up-regulated by hemin, heme and cobalt-protoporphyrin ameliorates experimental colitis. In addition, the up-regulation of HO-1 partially explains the mechanism of action of 5-aminosalicylic acid (5-ASA), which is used clinically as an anti-colitis agent. In 2009 Ueda et al. has reported for the first time that mucosal protection by Polaprezinc, a chelate compound of zinc and L-carnosine used as an anti-ulcer drug in Japan, is also attributed to induction of HO-1 in the stomach. Since then, inducers of HO-1 are desired subject of research, as they may constitute therapeutically effective anti-ulcer drugs.

Keywords: heme oxygenase-1, gastric lesions, gastroprotection, Polaprezinc

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Authors: N. C. Okoronkwo, I. E. Mbaeyi-Nwaoha, C. P. Agbata

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Childhood malnutrition is one of the most persistent public health problems throughout developing countries, including Nigeria. Demographic and Health survey data from twenty-one developing countries indicated that poor complementary feeding of children aged 6- 23 months contributes to negative growth trends. To reduce malnutrition among children in the society, formulation of complimentary food rich in essential nutrient for optimum growth and development of infants is essential. This study focused on the evaluation of complementary food produced by solid-state fermentation of Acha and Soybean using Rhizopus oligosporus (2710) and Orange-fleshed sweet potatoes (OFSP) using Lactobacillus planterum (B-41621). The raw materials were soaked separately, each in four volumes of 0.9M acetic acid for 16 hours, rinsed with clean water, steam cooked and cooled. Solid-state fermentation (SSF) was carried out by inoculating Acha and Soybean with spore suspension (1x 10⁶spores/ml) of Rhizopus oligosporus (2710) and OFSP with spore suspension (1x 106spores/ml) of Lactobacillus planterum (B-41621). Fermentation which lasted for 72hours was carried out with 24hours sampling. The samples were blended in the following ratios: Acha and soybean 100: 100 (AS), Acha/soybean and OFSP 50: 50(ASO), made into gruel and compared with a commercial infant formula (Cerelac) which served as the control (CTRL). The samples were analyzed for proximate composition using AOAC methods and sensory attributes using a hedonic scale. Results showed that moisture, crude protein, fibre and ash content increased significantly (p<0.05) as fermentation progressed, while carbohydrate and fat content decreased. The protein, moisture, fibre and ash content ranged from 17.10-19.02%, 54.97-56.27%, 7.08-7.60% and2.09-2.38%, respectively, while carbohydrate and fat content ranged from 12.95-10.21% and 5.81-4.52%, respectively. In sensory scores, there were no significant (p>0.05) difference between the average mean scores of colours, texture and consistency of the samples. The sensory score for the overall acceptability ranged from 6.20-7.80. Sample CTRL had the highest score, while sample ASO had the least score. There was no significant (p>0.05) difference between samples CTRL and AS. Solid-state fermentation improved the nutritional content and flavour of the developed complementary food, which is needed for infant growth and development.

Keywords: Complementary food, malnutrition, proximate composition, solid-state fermentation

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Authors: Abdulnasir Hossen, Alaa Barhoum, Deepali Jaju, V. Gowri, L. Al-Kharusi, M. Hassan, K. Al-Hashmi

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Preeclampsia is a pregnancy disorder associated with increase in blood pressure and excess amount of protein in the urine. HRV analysis has been used by many researchers to identify preeclamptic pregnancy from normal pregnancy. A study in this regard to identify preeclamptic pregnancy in Oman from normal pregnant was conducted on 40 subjects (20 patients and 20 normal). The subjects were collected from two hospitals in Oman. A Fast Fourier transform (FFT) spectral analysis has shown that patients with preeclamptic pregnancy have a reduction in the power of the HF band and an increase in the power of the LF band of HRV compared with subjects with normal pregnancy. The accuracy of identification obtained was 80%.

Keywords: preelampsia, pregnancy hypertension, normal pregnant, FFT, spectral analysis, HRV

Procedia PDF Downloads 539