Search results for: replication protein A

Authors: Yonggyu Lee, Byungkyu Jung, Bom Yoon, Jongil Yoon

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MR-based simulation devices have been recently used in various fields such as entertainment, medicine, manufacturing, and education. Different simulation devices are also being developed for military equipment training. This is to address the concerns regarding safety accidents as well as cost issues associated with training with expensive equipment. An important aspect of developing simulation devices to replicate military training is that trainees experience the same effect as training with real devices. In this study, the criteria for performance evaluation are established to compare the training effect of an MR-based simulation device to that of an actual device. K-9 Self-propelled artillery (SPA) operators are selected as training subjects. First, MR-based software is developed to simulate the training ground and training scenarios currently used for training SPA operators in South Korea. Hardware that replicates the interior of SPA is designed, and a simulation device that is linked to the software is developed. Second, criteria are established to evaluate the simulation device based on real-life training scenarios. A total of nine performance evaluation criteria were selected based on the actual SPA operation training scenarios. Evaluation items were selected to evaluate whether the simulation device was designed such that trainees would experience the same effect as training in the field with a real SPA. To eval-uate the level of replication by the simulation device of the actual training environments (driving and passing through trenches, pools, protrusions, vertical obstacles, and slopes) and driving conditions (rapid steering, rapid accelerating, and rapid braking) as per the training scenarios, tests were performed under the actual training conditions and in the simulation device, followed by the comparison of the results. In addition, the level of noise felt by operators during training was also selected as an evaluation criterion. Due to the nature of the simulation device, there may be data latency between HW and SW. If the la-tency in data transmission is significant, the VR image information delivered to trainees as they maneuver HW might not be consistent. This latency in data transmission was also selected as an evaluation criterion to improve the effectiveness of the training. Through this study, the key evaluation metrics were selected to achieve the same training effect as training with real equipment in a training ground during the develop-ment of the simulation device for military equipment training.

Keywords: K-9 self-propelled artillery, mixed reality, simulation device, synchronization

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Authors: Sourav Ray, Christoph Stein, Marcus Weber

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A new class of drug candidates, initially derived from mathematical modeling of ligand-receptor interactions, activate the μ-opioid receptor (MOR) preferentially at acidic extracellular pH-levels, as present in injured tissues. This is of commercial interest because it may preclude the adverse effects of conventional MOR agonists like fentanyl, which include but are not limited to addiction, constipation, sedation, and apnea. Animal studies indicate the importance of taking the pH value of the chemical environment of MOR into account when designing new drugs. Hydrogen bonds (HBs) play a crucial role in stabilizing protein secondary structure and molecular interaction, such as ligand-protein interaction. These bonds may depend on the pH value of the chemical environment. For the MOR, antagonist naloxone and agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) form HBs with ionizable residue HIS 297 at physiological pH to modulate signaling. However, such interactions were markedly reduced at acidic pH. Although fentanyl-induced signaling is also diminished at acidic pH, HBs with HIS 297 residue are not observed at either acidic or physiological pH for this strong agonist of the MOR. Molecular dynamics (MD) simulations can provide greater insight into the interaction between the ligand of interest and the HIS 297 residue. Amino acid protonation states are adjusted to the model difference in system acidity. Unbiased and unrestrained MD simulations were performed, with the ligand in the proximity of the HIS 297 residue. Ligand-receptor complexes were embedded in 1-palmitoyl-2-oleoyl-sn glycero-3-phosphatidylcholine (POPC) bilayer to mimic the membrane environment. The occurrence of HBs between the different ligands and the HIS 297 residue of MOR at acidic and physiological pH values were tracked across the various simulation trajectories. No HB formation was observed between fentanyl and HIS 297 residue at either acidic or physiological pH. Naloxone formed some HBs with HIS 297 at pH 5, but no such HBs were noted at pH 7. Interestingly, DAMGO displayed an opposite yet more pronounced HB formation trend compared to naloxone. Whereas a marginal number of HBs could be observed at even pH 5, HBs with HIS 297 were more stable and widely present at pH 7. The HB formation plays no and marginal role in the interaction of fentanyl and naloxone, respectively, with the HIS 297 residue of MOR. However, HBs play a significant role in the DAMGO and HIS 297 interaction. Post DAMGO administration, these HBs might be crucial for the remediation of opioid tolerance and restoration of opioid sensitivity. Although experimental studies concur with our observations regarding the influence of HB formation on the fentanyl and DAMGO interaction with HIS 297, the same could not be conclusively stated for naloxone. Therefore, some other supplementary interactions might be responsible for the modulation of the MOR activity by naloxone binding at pH 7 but not at pH 5. Further elucidation of the mechanism of naloxone action on the MOR could assist in the formulation of cost-effective naloxone-based treatment of opioid overdose or opioid-induced side effects.

Keywords: effect of system acidity, hydrogen bond formation, opioid action, receptor activation

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Authors: Janet Modu, Jean-Francois Georgin, Laurent Briancon, Eric Antoinet

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The current practice during the repowering phase of wind turbines is deconstruction of existing foundations and construction of new foundations to accept larger wind loads or once the foundations have reached the end of their service lives. The ongoing research project FUI25 FEDRE (Fondations d’Eoliennes Durables et REpowering) therefore serves to propose scalable wind turbine foundation designs to allow reuse of the existing foundations. To undertake this research, numerical models and laboratory-scale models are currently being utilized and implemented in the GEOMAS laboratory at INSA Lyon following instrumentation of a reference wind turbine situated in the Northern part of France. Sensors placed within both the foundation and the underlying soil monitor the evolution of stresses from the foundation’s early age to stresses during service. The results from the instrumentation form the basis of validation for both the laboratory and numerical works conducted throughout the project duration. The study currently focuses on the effect of coupled mechanisms (Thermal-Hydro-Mechanical-Chemical) that induce stress during the early age of the reinforced concrete foundation, and scale factor considerations in the replication of the reference wind turbine foundation at laboratory-scale. Using THMC 3D models on COMSOL Multi-physics software, the numerical analysis performed on both the laboratory-scale and the full-scale foundations simulate the thermal deformation, hydration, shrinkage (desiccation and autogenous) and creep so as to predict the initial damage caused by internal processes during concrete setting and hardening. Results show a prominent effect of early age properties on the damage potential in full-scale wind turbine foundations. However, a prediction of the damage potential at laboratory scale shows significant differences in early age stresses in comparison to the full-scale model depending on the spatial position in the foundation. In addition to the well-known size effect phenomenon, these differences may contribute to inaccuracies encountered when predicting ultimate deformations of the on-site foundation using laboratory scale models.

Keywords: cement hydration, early age behavior, reinforced concrete, shrinkage, THMC 3D models, wind turbines

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Authors: L. Strauta, S. Muižniece-Brasava

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Even though legumes possess high nutritional value and have a rather high protein content for plant origin products, they are underutilized mostly due to their lengthy cooking time. To increase the presence of legume-based products in human diet, new extruded products were made of cow peas (Vigna unguiculata (L.) Walp.). But as it is known, adding different moisture content to flour before extrusion can change the physical properties of the extruded product. Experiments were carried out to estimate the optimal moisture content for cow pea extrusion. After extrusion, the pH level had dropped from 6.7 to 6.5 and the lowest hardness rate was observed in the samples with additional 9 g 100g^-1 of moisture - 28±4N, but the volume mass of the samples with additional 9 g100g^-1 of water was 263±3 g L^-1; all samples were approximately 7±1mm long.

Keywords: cow pea, extrusion–cooking, moisture, size

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Authors: Vilma Sasyte, Vilma Viliene, Asta Raceviciute-Stupeliene, Agila Dauksiene, Romas Gruzauskas, Virginijus Slausgalvis, Jamal Al-Saifi

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The aim of the present study was to investigate the effect of butyrate, acids, medium-chain fatty acids and plant extract mixture on performance, blood and gastrointestinal tract characteristics of laying hens’. For the period of 8 weeks, 24 Hisex Brown laying hens were randomly assigned to 2 dietary treatments: 1) control wheat-corn-soybean meal based diet (Control group), 2) control diet supplemented with the mixture of butyrate, acids, medium chain fatty acids and plant extract (Lumance®) at the level of 1.5 g/kg of feed (Experimental group). Hens were fed with a crumbled diet at 125 g per day. Housing and feeding conditions were the same for all groups and met the requirements of growth for laying hens of Hisex Brown strain. In the blood serum total protein, bilirubin, cholesterol, DTL- and MTL- cholesterol, triglycerides, glucose, GGT, GOT, GPT, alkaline phosphatase, alpha amylase, contents of c-reactive protein, uric acid, and lipase were analyzed. Development of intestines and internal organs (intestinal length, intestinal weight, the weight of glandular and muscular stomach, pancreas, heart, and liver) were determined. The concentration of short chain fatty acids in caecal content was measured using the method of HPLC. The results of the present study showed that 1.5 g/kg supplementation of feed additive affected egg production and feed conversion ratio for the production of 1 kg of egg mass. Dietary supplementation of analyzed additive in the diets increased the concentration of triglycerides, GOT, alkaline phosphatase and decreased uric acid content compared with the control group (P<0.05). No significant difference for others blood indices in comparison to the control was observed. The addition of feed additives in laying hens’ diets increased intestinal weight by 11% and liver weight by 14% compared with the control group (P<0.05). The short chain fatty acids (propionic, acetic and butyric acids) in the caecum of laying hens in experimental groups decreased compared with the control group. The supplementation of the mixture of butyrate, acids, medium-chain fatty acids and plant extract at the level of 1.5 g/kg in the laying hens’ diets had the effect on the performance, some gastrointestinal tract function and blood parameters of laying hens.

Keywords: acids, butyrate, laying hens, MCFA, performance, plant extract, psysiological state

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Authors: L. García, N. Cobas, M. López

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Acrylamide, a probable carcinogen, is formed in high-temperature processed food (>120ºC) when the free amino acid asparagine reacts with reducing sugars, mainly glucose and fructose. Cane juices' repeated heating would potentially form acrylamide during brown sugar production. This study aims to determine if using panela in yogurt cake preparation increases acrylamide formation. A secondary aim is to analyze the acrylamide concentration in four cake confections with different caloric sweetener ingredients: beet sugar (BS), cane sugar (CS), panela (P), and a panela and chocolate mix (PC). The doughs were obtained by combining ingredients in a planetary mixer. A model system made up of flour (25%), caloric sweeteners (25 %), eggs (23%), yogurt (15.7%), sunflower oil (9.4%), and brewer's yeast (2 %) was applied to BS, CS and P cakes. The ingredients of PC cakes varied: flour (21.5 %), panela chocolate (21.5 %), eggs (25.9 %), yogurt (18 %), sunflower oil (10.8 %), and brewer’s yeast (2.3 %). The preparations were baked for 45' at 180 ºC. Moisture was estimated by AOAC. Protein was determined by the Kjeldahl method. Ash percentage was calculated by weight loss after pyrolysis (≈ 600 °C). Fat content was measured using liquid-solid extraction in hydrolyzed raw ingredients and final confections. Carbohydrates were determined by difference and total sugars by the Luff-Schoorl method, based on the iodometric determination of copper ions. Finally, acrylamide content was determined by LC-MS by the isocratic system (phase A: 97.5 % water with 0.1% formic acid; phase B: 2.5 % methanol), using a standard internal procedure. Statistical analysis was performed using SPSS v.23. One-way variance analysis determined differences between acrylamide content and compositional analysis, with caloric sweeteners as fixed effect. Significance levels were determined by applying Duncan's t-test (p<0.05). P cakes showed a lower energy value than the other baked products; sugar content was similar to BS and CS, with 6.1 % mean crude protein. Acrylamide content in caloric sweeteners was similar to previously reported values. However, P and PC showed significantly higher concentrations, probably explained by the applied procedure. Acrylamide formation depends on both reducing sugars and asparagine concentration and availability. Beet sugar samples did not present acrylamide concentrations within the detection and quantification limit. However, the highest acrylamide content was measured in the BS. This may be due to the higher concentration of reducing sugars and asparagine in other raw ingredients. The cakes made with panela, cane sugar, or panela with chocolate did not differ in acrylamide content. The lack of asparagine measures constitutes a limitation. Cakes made with panela showed lower acrylamide formation than products elaborated with beet or cane sugar.

Keywords: beet sugar, cane sugar, panela, yogurt cake

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Authors: Wirot Likittrakulwong, Pisit Poolprasert, Tossaporn Incharoen

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Germinated paddy rice (GPR) has the potential to be used as a feed ingredient. However, their properties have not been fully investigated. This paper examined the nutrient digestibility and the relationship to immune activity in Roman hens fed with GPR. It was found that true and apparent metabolizable energy (ME) values of GPR were 3.20 and 3.28 kcal/g air dry, respectively. GPR exhibited high content of phytonutrients, especially GABA. GPR showed similar protein profiles in comparison to non-germinated paddy rice. For immune activity, the feed with GPR enhanced the immune activity of Roman hens under high stocking density stress as evidenced by the activity of superoxide dismutase (SOD) and lysozyme activity. In this study, GPR is proved to be a good source of functional ingredient for chicken feed.

Keywords: germinated paddy rice, nutrient digestibility, immune activity, functional property

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Authors: Vanessa Funk, Steven Blum, Stephanie Cole, Jorge Maciel, Matthew Lux

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Paper-based synthetic gene circuits offer a new paradigm for programmable, fieldable biodetection. We demonstrate that by freeze-drying gene circuits with in vitro expression machinery, we can use complimentary RNA sequences to trigger colorimetric changes upon rehydration. We have successfully utilized both green fluorescent protein and luciferase-based reporters for easy visualization purposes in solution. Through several efforts, we are aiming to use this new platform technology to address a variety of needs in portable detection by demonstrating several more expression and reporter systems for detection functions on paper. In addition to RNA-based biodetection, we are exploring the use of various mechanisms that cells use to respond to environmental conditions to move towards all-hazards detection. Examples include explosives, heavy metals for water quality, and toxic chemicals.

Keywords: cell-free lysates, detection, gene circuits, in vitro

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Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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Authors: Nouha Bouayed Abdelmoula, Rim Louati, Nawel Abdellaoui, Balkiss Abdelmoula, Oldez Kaabi, Walid Smaoui, Samir Aloulou

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Background and Aims: Cardiofaciocutaneous syndrome (CFC) is an autosomal dominant disorder with the vast majority of cases arising by a new mutation of BRAF, MEK1, MEK2, or rarely, KRAS genes. Here, we report a rare Tunisian case of CFC syndrome for whom we identify SOS1 mutation. Methods: Genomic DNA was obtained from peripheral blood collected in an EDTA tube and extracted from leukocytes using the phenol/chloroform method according to standard protocols. High resolution melting (HRM) analysis for screening of mutations in the entire coding sequence of PTPN11 was conducted first. Then, HRM assays to look for hot spot mutations coding regions of the other genes of the RAS-MAPK pathway (RAt Sarcoma viral oncogene homolog Mitogen-Activated Protein Kinases Pathway): SOS1, SHOC2, KRAS, RAF1, KRAS, NRAS, CBL, BRAF, MEK1, MEK2, HRAS, and RIT1, were applied. Results: Heterozygous SOS1 point mutation clustered in exon 10, which encodes for the PH domain of SOS1, was identified: c.1655 G > A. The patient was a 9-year-old female born from a consanguineous couple. She exhibited pulmonic valvular stenosis as congenital heart disease. She had facial features and other malformations of Noonan syndrome, including macrocephaly, hypertelorism, ptosis, downslanting palpebral fissures, sparse eyebrows, a short and broad nose with upturned tip, low-set ears, high forehead commonly associated with bitemporal narrowing and prominent supraorbital ridges, short and/or webbed neck and short stature. However, the phenotype is also suggestive of CFC syndrome with the presence of more severe ectodermal abnormalities, including curly hair, keloid scars, hyperkeratotic skin, deep plantar creases, and delayed permanent dentition with agenesis of the right maxillary first molar. Moreover, the familial history of the patient revealed recurrent brain malignancies in the paternal family and epileptic disease in the maternal family. Conclusions: This case report of an overlapping RASopathy associated with SOS1 mutation and familial history of brain tumorigenesis is exceptional. The evidence suggests that RASopathies are truly cancer-prone syndromes, but the magnitude of the cancer risk and the types of cancer partially overlap.

Keywords: cardiofaciocutaneous syndrome, CFC, SOS1, brain cancer, germline mutation

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Authors: M. Iqbal Hossain, Azharul Islam Khan, S. M. Rafiqul Islam, Tahmeed Ahmed

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Objective: Moderate acute malnutrition (MAM) is a major cause of morbidity and mortality in under-5 children of low-income countries. Approximately 14.6% of all under-5 mortality worldwide is attributed to MAM with >3 times increased risk of death compared to well-nourished peers. Prevalence of MAM among under-5 children in Bangladesh is ~12% (~1.7 million). Providing a diet containing adequate nutrients is the mainstay of treatment of children with MAM. It is now possible to process fish into fish peptides with longer shelf-life without refrigerator, known as ‘Fish Surimi peptide’ and this could be an attractive alternative to supply fish protein in the diet of children in low-income countries like Bangladesh. We conducted this study to assess the acceptability of Fish Surimi peptide given with various foods/meals in 2-5 years old children with MAM. Design/methods: Fish Surimi peptide is broken down from white fish meat using plant-derived enzyme and the ingredient is just fish meat consisted of 20 different kinds of amino acids including nine essential amino acids. In a convenience sample of 34 children we completed the study ward of Dhaka Hospital of icddr,b in Bangladesh during November 2014 through February 2015. For each child the study was for two consecutive days: i.e. direct observation of food intake of two lunches and two suppers. In a randomly and blinded manner and cross over design an individual child received Fish Surimi peptide (5g at lunch and 5g at supper) mixed meal [e.g. 30g rice and 30g dahl (thick lentil soup) or 60g of a vegetables-lentil-rice mixed local dish known as khichuri in one day and the same meal on other day without any Fish Surimi peptide. We observed the completeness and eagerness of eating and any possible side effect (e.g. allergy, vomiting, diarrhea etc.) over these two days. Results: The mean±SD age of the enrolled children was 38.4±9.4 months, weight 11.22±1.41 kg, height 91.0±6.3 cm, and WHZ was -2.13±0.76. Their mean±SD total feeding time (minutes) for lunch was 25.4±13.6 vs. 20.6±11.1 (p=0.130) and supper was 22.3±9.7 vs. 19.7±11.2 (p=0.297), and total amount (g) of food eaten in lunch and supper was found similar 116.1±7.0 vs. 117.7±8.0 (p=3.01) in A (Fish Surimi) and B group respectively. Score in Hedonic scale by mother on test of food given to children at lunch or supper was 3.9±0.2 vs. 4.0±0.2 (p=0.317) and on overall acceptance (including the texture, smell, and appearance) of food at lunch or supper was 3.9±0.2 vs. 4.0±0.2 (p=0.317) for A and B group respectively. No adverse event was observed in any food group during the study period. Conclusions: Fish Surimi peptide may be a cost effective supplementary food, which should be tested by appropriately designed randomized community level intervention trial both in wasted children and stunted children.

Keywords: protein-energy malnutrition, moderate acute malnutrition, weight-for-height z-score, mid upper arm circumference, acceptability, fish surimi peptide, under-5 children

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Authors: Hategekimana Jean Paul, Irakoze Josiane, Ishimweyizerwe Valentin, Iradukunda Dieudonne, Uwanyirigira Jeannette

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Orange fleshed sweet potatoes (OFSP) are highly grown and are available plenty in rural and urban local markets and its contribution in reduction of food insecurity in Rwanda is considerable. But the postharvest loss of this commodity is a critical challenge due to its high perishability. Several research activities have been conducted on how fresh food commodities can be transformed into extended shelf life food products for prevention of post-harvest losses. However, such activity was not yet well studied in Rwanda. The aim of the present study was the processing of backed products from (OFSP)combined with wheat composite flour and assess the nutritional content and consumer acceptability of new developed products. The perishability of OFSP and their related lack during off season can be eradicated by producing cake, doughnut and bread with OFSP puree or flour. The processing for doughnut and bread were made by making OFSP puree and other ingredients then a dough was made followed by frying and baking while for cake OFSP was dried through solar dryer to have a flour together with wheat flour and other ingredients to make dough cake and baking. For each product, one control and three experimental samples, (three products in three different ratios (30,40 and50%) of OFSP and the remaining percentage of wheat flour) were prepared. All samples including the control were analyzed for the consumer acceptability (sensory attributes). Most preferred samples (One sample for each product with its control sample and for each OFSP variety) were analyzed for nutritional composition along with control sample. The Cake from Terimbere variety and Bread from Gihingumukungu supplemented with 50% OFSP flour or Puree respectively were most acceptable except Doughnut from Vita variety which was highly accepted at 50% of OFSP supplementation. The moisture, ash, protein, fat, fiber, Total carbohydrate, Vitamin C, reducing sugar and minerals (Sodium, Potassium and Phosphorus.) content was different among products. Cake was rich in fibers (14.71%), protein (6.590%), and vitamin c(19.988mg/100g) compared to other samples while bread found to be rich in reducing sugar with 12.71mg/100g compared to cake and doughnut. Also doughnut was found to be rich in fat content with 6.89% compared to other samples. For sensory analysis, doughnut was highly accepted in ratio of 60:40 compared to other products while cake was least accepted at ratio of 50:50. The Proximate composition and minerals content of all the OFSP products were significantly higher as compared to the control samples.

Keywords: post-harvest loss, OFSP products, wheat flour, sensory evaluation, proximate composition

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Authors: V. Villa-Cruz, H. Pérez-Ladron de Guevara, J. E. Diaz-Díaz

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Static Light Scattering (SLS) was used as a method to analyse cow's milk raw, coming from the town of Lagos de Moreno, Jalisco, Mexico. This method is based on the analysis of the dispersion of light laser produced by a set of particles in solution. Based on the above, raw milk, which contains particles of fat globules, with a diameter of 2000 nm and particles of micelles of protein with 300 nm in diameter were analyzed. For this, dilutions of commercial milk were made (1.0%, 2.0% and 3.3%) to obtain a pattern of laser light scattering and also made measurements of raw cow's milk. Readings were taken in a sweep initial angle 10° to 170°, results were analyzed with the program OriginPro 7. The SLS method gives us an estimate of the percentage of fat content in milk samples. It can be concluded that the SLS method, is a quick method of analysis to detect adulteration in raw cow's milk.

Keywords: light scattering, milk analysis, adulteration in milk, micelles, OriginPro

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Authors: Ahmed M. Eed, Adam H. Burgoyne

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Jojoba (Simmondsia chinensis (Link) Schneider), is a desert shrub which tolerates saline, alkyle soils and drought. The seeds contain a characteristic liquid wax of economic importance in industry as a machine lubricant and cosmetics. A major problem in seed propagation is that jojoba is a dioecious plant whose sex is not easily determined prior to flowering (3-4 years from germination). To overcome this phenomenon, asexual propagation using vegetative methods such as cutting can be used. This research was conducted to find out the effect of different Plant Growth Regulators (PGRs) and rooting media on Jojoba rhizogenesis. An experiment was carried out in a Factorial Completely Randomized Design (FCRD) with three replications, each with sixty cuttings per replication in fiberglass house of Natural Jojoba Corporation at Yemen. The different rooting media used were peat moss + perlite + vermiculite (1:1:1), peat moss + perlite (1:1) and peat moss + sand (1:1). Plant materials used were semi-hard wood cuttings of jojoba plants with length of 15 cm. The cuttings were collected in the month of June during 2012 and 2013 from the sub-terminal growth of the mother plants of Amman farm and introduced to Yemen. They were wounded, treated with Indole butyric acid (IBA), α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) all @ 4000 ppm (part per million) and cultured on different rooting media under intermittent mist propagation conditions. IBA gave significantly higher percentage of rooting (66.23%) compared to NAA and IAA in all media used. However, the lowest percentage of rooting (5.33%) was recorded with IAA in the medium consisting of peat moss and sand (1:1). No significant difference was observed at all types of PGRs used with rooting media in respect of root length. Maximum number of roots was noticed in medium consisting of peat moss, perlite and vermiculite (1:1:1); peat moss and perlite (1:1) and peat moss and sand (1:1) using IBA, NAA and IBA, respectively. The interaction among rooting media was statistically significant with respect to rooting percentage character. Similarly, the interactions among PGRs were significant in terms of rooting percentage and also root length characters. The results demonstrated suitability of propagation of jojoba plants by semi-hard wood cuttings.

Keywords: cutting, IBA, Jojoba, propagation, rhizogenesis

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Authors: Asim Cengiz, Dede Basturk, Hakan Ozalp

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Practicing recurring resistance workout such as may cause changes in human muscle. These changes may be because combination if several factors determining physical fitness. Thus, it is important to identify these changes. Several studies were reviewed to investigate these changes. As a result, the changes included positive modifications in amplified citrate synthase (CS) maximal activity, increased capacity for pyruvate oxidation, improvement on molecular signaling on human performance, amplified resting muscle glycogen and whole GLUT4 protein content, better health outcomes such as enhancement in cardiorespiratory fitness. Sprint training also have numerous long long-term changes inhuman body such as better enzyme action, changes in muscle fiber and oxidative ability. This is important because SV is the critical factor influencing maximal cardiac output and therefore oxygen delivery and maximal aerobic power.

Keywords: sprint, training, performance, exercise

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Authors: Ashleigh Hillier, Jody Goldstein, Lauren Tornatore, Emily Byrne

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As individuals with disabilities attend higher education in greater numbers, universities are seeking ways to support the retention and success of these students, beyond the academically based accommodations. Although mentoring programs for this population are being implemented more frequently, there is a lack of empirically validated outcomes which could promote program replication. The research objective of this exploratory study was to examine outcomes for students with disabilities participating in a peer-to-peer mentoring program. Mentees (students with disabilities) met with their mentor (trained upperclassman) once a week for an hour for one semester (14-weeks). Mentors followed a curriculum structured by monthly and weekly goals to guide the sessions. Curriculum topics included socializing on campus, peer pressure, time management, communicating with peers and professors, classroom etiquette, study skills, and seeking help and campus resources. Data was collected over a period of seven semesters resulting in seven separate cohorts (n=46). The impact of the program was measured using quantitative self-report measures as well as qualitative content analysis of focus groups. Academic outcomes (retention, credits earned, and GPA) were compared between those in the mentoring program and a matched group of students registered with Disability Services who did not receive mentoring. In addition, a one-year follow up was conducted to examine the longer term impact of participation. Findings indicated that mentoring had the most impact in knowing how things work at the university, knowing how and where to find opportunities to meet people on campus, and knowing how to access supports. Mentors also provided a supportive relationship to the mentees and helped with social skills. There were no significant differences in academic outcomes between those who were mentored and those in the comparison group. Most mentees reported continuing to benefit from the program one year on, providing support for the retention of knowledge gained and maintenance of positive outcomes over time. In conclusion, while a range of positive outcomes were evidenced, the model was limited in its impact more broadly, particularly with regards to academic success and impacting more complex challenges.

Keywords: mentor, outcomes, students with disabilities, university

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Authors: Mohamed Fawzy Elzarei, Yousef Mohammed Al-Dakheel, Ali Mohamed Alseaf

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Modern molecular techniques, like single marker analysis for linked traits to these markers, can provide us with rapid and accurate genetic results. In the last two decades of the last century, the applications of molecular techniques were reached a faraway point in cattle, sheep, and pig. In goats, especially in our region, the application of molecular techniques is still far from other species. As reported by many researchers, microsatellites marker is one of the suitable markers for lie studies. The single marker linked to traits of interest is one technique allowed us to early select animals without the necessity for mapping the entire genome. Simplicity, applicability, and low cost of this technique gave this technique a wide range of applications in many areas of genetics and molecular biology. Also, this technique provides a useful approach for evaluating genetic differentiation, particularly in populations that are poorly known genetically. The expected breeding value (EBV) and yield deviation (YD) are considered as the most parameters used for studying the linkage between quantitative characteristics and molecular markers, since these values are raw data corrected for the non-genetic factors. A total of 17 microsatellites markers (from chromosomes 6, 14, 18, 20 and 23) were used in this study to search for areas that could be responsible for genetic variability for some milk traits and search of chromosomal regions that explain part of the phenotypic variance. Results of single-marker analyses were used to identify the linkage between microsatellite markers and variation in EBVs of these traits, Milk yield, Protein percentage, Fat percentage, Litter size and weight at birth, and litter size and weight at weaning. The estimates of the parameters from forward and backward solutions using stepwise regression procedure on milk yield trait, only two markers, OARCP9 and AGLA29, showed a highly significant effect (p≤0.01) in backward and forward solutions. The forward solution for different equations conducted that R2 of these equations were highly depending on only two partials regressions coefficient (βi,) for these markers. For the milk protein trait, four marker showed significant effect BMS2361, CSSM66 (p≤0.01), BMS2626, and OARCP9 (p≤0.05). By the other way, four markers (MCM147, BM1225, INRA006, andINRA133) showed highly significant effect (p≤0.01) in both backward and forward solutions in association with milk fat trait. For both litter size at birth and at weaning traits, only one marker (BM143(p≤0.01) and RJH1 (p≤0.05), respectively) showed a significant effect in backward and forward solutions. The estimates of the parameters from forward and backward solution using stepwise regression procedure on litter weight at birth (LWB) trait only one marker (MCM147) showed highly significant effect (p≤0.01) and two marker (ILSTS011, CSSM66) showed a significant effect (p≤0.05) in backward and forward solutions.

Keywords: microsatellites marker, estimated breeding value, stepwise regression, milk traits

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Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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Authors: Shasha Lv, Zhengcao Li

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Due to the CMOS compatibility, silicon-based field emission (FE) devices as potential electron sources have attracted much attention. The geometrical arrangement and dimensional features of aligned silicon nanowires (SiNWs) have a determining influence on the FE properties. We discuss a multistep template replication process of Ag-assisted chemical etching combined with polystyrene (PS) spheres to fabricate highly periodic and well-aligned silicon nanowires, then their diameter, aspect ratio and density were further controlled via dry oxidation and post chemical treatment. The FE properties related to proximity and aspect ratio were systematically studied. A remarkable improvement of FE propertiy was observed with the average nanowires tip interspace increasing from 80 to 820 nm. On the basis of adjusting SiNWs dimensions and morphology, addition of a secondary material whose properties complement the SiNWs could yield a combined characteristic. Three different nanoheterostructures were fabricated to control the FE performance, they are: NiSi/Si core-shell structures, ZnO/Si nanotrees, and Graphene/SiNWs. We successfully fabricated the high-quality NiSi/Si heterostructured nanowires with excellent conformality. First, nickle nanoparticles were deposited onto SiNWs, then rapid thermal annealing process were utilized to form NiSi shell. In addition, we demonstrate a new and simple method for creating 3D nanotree-like ZnO/Si nanocomposites with a spatially branched hierarchical structure. Compared with the as-prepared SiNRs and ZnO NWs, the high-density ZnO NWs on SiNRs have exhibited predominant FE characteristics, and the FE enhancement factors were attributed to band bending effect and geometrical morphology. The FE efficiency from flat sheet structure of graphene is low. We discussed an effective approach towards full control over the diameter of uniform SiNWs to adjust the protrusions of large-scale graphene sheet deposited on SiNWs. The FE performance regarding the uniformity and dimensional control of graphene protrusions supported on SiNWs was systematically clarified. Therefore, the hybrid SiNWs/graphene structures with protrusions provide a promising class of field emission cathodes.

Keywords: field emission, silicon nanowires, heterostructures, controllable synthesis

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Authors: Syed Sameer Aga, Saniya Nissar

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The Xeroderma pigmentosum group D gene (XPD) plays a key role in nucleotide excision repair (NER) pathway of the damaged DNA. Genetic polymorphisms in the coding region of the XPD gene may alter DNA repair capacity of the protein and hence can modulate the risk of colorectal cancer (CRC) risk. The aim of the study was to determine the genetic association of XPD Lys751Gln polymorphism with the risk of colorectal cancer (CRC) development. 120 CRC patients and 160 normal controls were assessed for genotype frequencies of XPD Lys751Gln polymorphism using PCR-RFLP technique. We observed a significant association (p < 0.05) between the XPD Lys751Gln polymorphism and the risk of developing CRC (p < 0.05). Additionally, Gln/Gln genotype of the XPD gene doubled the risk for the development of CRC [p < 0.05; OR=2.25 95% CI (1.07-4.7)]. Our results suggest that there is a significant association between the XPD Lys751Gln polymorphism and the risk of CRC.

Keywords: colorectal cancer, polymorphism, RFLP, DNA Repair, NER, XPD

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Authors: P. Vosynek, T. Návrat, M. Peč, J. Pořízka, P. Diviš

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Changes of mechanical properties and response behavior of bones is an important external sign of medical problems like osteoporosis, bone remodeling after fracture or surgery, osteointegration, or bone tissue loss of astronauts in space. Measuring of mechanical behavior of bones in physiological and osteoporotic states, quantified by different degrees of protein (collagen) and mineral loss, is thus an important topic in biomechanical research. This contribution deals with the relation between mechanical properties of the turkey long bone–tibia in physiological, demineralized, and deproteinized state. Three methods for comparison were used: densitometry, three point bending and harmonic response analysis. The results help to find correlations between the methods and estimate their possible application in medical practice.

Keywords: bone properties, long bone, osteoporosis, response behavior

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Authors: Zeina Bou Diab, Arij Daou

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The neural mechanisms of sequential behaviors are intensively studied, with songbirds a focus for learned vocal production. We are studying the premotor nucleus HVC at a nexus of multiple pathways contributing to song learning and production. The HVC consists of multiple classes of neuronal populations, each has its own cellular, electrophysiological and functional properties. During singing, a large subset of motor cortex analog-projecting HVCRA neurons emit a single 6-10 ms burst of spikes at the same time during each rendition of song, a large subset of basal ganglia-projecting HVCX neurons fire 1 to 4 bursts that are similarly time locked to vocalizations, while HVCINT neurons fire tonically at average high frequency throughout song with prominent modulations whose timing in relation to song remains unresolved. This opens the opportunity to define models relating explicit HVC circuitry to how these neurons work cooperatively to control learning and singing. We developed conductance-based Hodgkin-Huxley models for the three classes of HVC neurons (based on the ion channels previously identified from in vitro recordings) and connected them in several physiologically realistic networks (based on the known synaptic connectivity and specific glutaminergic and gabaergic pharmacology) via different architecture patterning scenarios with the aim to replicate the in vivo firing patterning behaviors. We are able, through these networks, to reproduce the in vivo behavior of each class of HVC neurons, as shown by the experimental recordings. The different network architectures developed highlight different mechanisms that might be contributing to the propagation of sequential neural activity (continuous or punctate) in the HVC and to the distinctive firing patterns that each class exhibits during singing. Examples of such possible mechanisms include: 1) post-inhibitory rebound in HVCX and their population patterns during singing, 2) different subclasses of HVCINT interacting via inhibitory-inhibitory loops, 3) mono-synaptic HVCX to HVCRA excitatory connectivity, and 4) structured many-to-one inhibitory synapses from interneurons to projection neurons, and others. Replication is only a preliminary step that must be followed by model prediction and testing.

Keywords: computational modeling, neural networks, temporal neural sequences, ionic currents, songbird

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Authors: Tharwat Ibrahim Rushdy, Warda Youssef Mohammed Morsy, Hanaa Ali Ahmed Elfeky

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Background: Intra-aortic balloon pump (IABP) is the first and the most commonly used mechanical circulatory support for patients with acute coronary syndromes and cardiogenic shock. Therefore, critical care nurses not only have to know how to monitor and operate the IABP, but also to provide interventions for preventing possible complications. Aim of the study: To assess nurses' knowledge and practices regarding care of patients connected to IABP at the ICUs of Cairo University Hospitals. Research design: A descriptive exploratory design was utilized. Sample: Convenience samples of 40 nurses were included in the current study. Setting: This study was carried out at the Intensive Care Units of Cairo University Hospitals. Tools of data collection: Three tools were developed, tested for clarity, and feasibility: a- Nurses' personal background sheet, b- IABP nurses' knowledge self-administered questionnaire, and c- IABP Nurses' practice observational checklist. Results: The majority of the studied sample had unsatisfactory knowledge and practice level (88% & 95%) respectively with a mean of 9.45+2.94 and 30.5+8.7, respectively. Unsatisfactory knowledge was found regarding description and physiological effects, nursing care, indications, contraindications, complications, weaning, and removal of IABP in percentage of 95%, 90%, 72.5%, and 57.5%, respectively, with a mean total knowledge score of 9.45 +2.94. In addition, unsatisfactory practice was found regarding about preparation and initiation of IABP therapy, nursing practice during therapy, weaning, and removal of IABP in percentages of (97.5%, 97.5%, and 90%), respectively. Finally, knowledge level was found to differ significantly in relation to gender (t = 2.46 at P ≤ 0.018). However, gender didn't play a role in relation to practice (t = 0.086 at P≤ 0.932). Conclusion: In spite of having vital role in assessment and management of critically ill patients, critical care nurses in the current study had in general unsatisfactory knowledge and practice regarding care of patients connected to IABP. Recommendation: updating knowledge and practice of ICU nurses through carrying out continuing educational programs about IABP; strict observation of nurses' practice when caring for patients connected to IABP and provision of guidance to correct of poor practices and replication of this study on larger probability sample selected from different geographical locations.

Keywords: knowledge, practice, intra-aortic balloon pump (IABP), ICU nurses, intensive care unit (ICU), introduction

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Authors: Chloe Richards, Adrian Delgado Ollero, Yan Delaure, Fiona Regan

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Growing concern about the natural environment has accelerated the search for non-toxic, but at the same time, economically reasonable, antifouling materials. Bioinspired surfaces, due to their nano and micro topographical antifouling capabilities, provide a hopeful approach to the design of novel antifouling surfaces. Biological organisms are known to have highly evolved and complex topographies, demonstrating antifouling potential, i.e. shark skin. Previous studies have examined the antifouling ability of topographic patterns, textures and roughness scales found on natural organisms. One of the mechanisms used to explain the adhesion of cells to a substrate is called attachment point theory. Here, the fouling organism experiences increased attachment where there are multiple attachment points and reduced attachment, where the number of attachment points are decreased. In this study, an attempt to characterize the microtopography of the common brill fish, Scophthalmus rhombus, was undertaken. Scophthalmus rhombus is a small flatfish of the family Scophthalmidae, inhabiting regions from Norway to the Mediterranean and the Black Sea. They reside in shallow sandy and muddy coastal areas at depths of around 70 – 80 meters. Six engineered surfaces (inspired by the Brill fish scale) produced by a 2-photon polymerization (2PP) process were evaluated for their potential as an antifouling solution for incorporation onto tidal energy blades. The micro-textures were analyzed for their AF potential under both static and dynamic laboratory conditions using two laboratory grown diatom species, Amphora coffeaeformis and Nitzschia ovalis. The incorporation of a surface topography was observed to cause a disruption in the growth of A. coffeaeformis and N. ovalis cells on the surface in comparison to control surfaces. This work has demonstrated the importance of understanding cell-surface interaction, in particular, topography for the design of novel antifouling technology. The study concluded that biofouling can be controlled by physical modification, and has contributed significant knowledge to the use of a successful novel bioinspired AF technology, based on Brill, for the first time.

Keywords: attachment point theory, biofouling, Scophthalmus rhombus, topography

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Authors: Petnamnueng Dettipponpong, Shuen E. Chen

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Heat stress is known to have negative effects on reproductive functions, such as follicular development and ovulation. This study aimed to investigate the specific effects of heat stress on steroid hormone secretion of ovarian follicle cells, particularly in relation to the expression of Apolipoprotein B (ApoB) and microsomal triglyceride transfer protein (MTP). The aim of the study was to understand the impact of heat stress on steroid hormone secretion in ovarian follicle cells and to explore the role of ApoB and MTP in this process. Primary granulosa and theca cells were collected from follicles and cultured under heat stress conditions (42 °C) for various time periods. Controls were maintained under normal conditions (37.5 °C ). The culture medium was collected at different time points to measure levels of progesterone and estradiol using ELISA kits. ApoB and MTP expression levels were analyzed using homemade antibodies and western blot. Data were assessed by a one-way ANOVA comparison test with Duncan’s new multiple-range test. Results were expressed as mean±S.E. Difference was considered significant at P<0.05. The results showed that heat stress significantly increased progesterone secretion in granulosa cells, with the peak observed after 13 hours of recovery under thermoneutral conditions. Estradiol secretion by theca cells was not affected. Heat stress also had a significant negative effect on granulosa cell viability. Additionally, the expression of ApoB and MTP was found to be differentially regulated by heat stress. ApoB expression in theca cells was transiently promoted, while ApoB expression in granulosa cells was consistently suppressed. MTP expression increased after 5 hours of recovery in both cell types. These findings suggest a mechanism by which chicken follicle cells export cellular lipids as very low-density lipoprotein (VLDL) in response to thermal stress. These contribute to our understanding of the role of ApoB and MTP steroidogenesis and lipid metabolism under heat stress conditions. The study involved the collection of primary granulosa and theca cells, culture under different temperature conditions, and analysis of the culture medium for hormone levels using ELISA kits. ApoB and MTP expression levels were assessed using homemade antibodies and western blot. This study aimed to address the effects of heat stress on steroid hormone secretion in ovarian follicle cells, as well as the role of ApoB and MTP in this process. The study demonstrates that heat stress stimulates steroidogenesis in granulosa cells, affecting progesterone secretion. ApoB and MTP expression were found to be differentially regulated by heat stress, indicating a potential mechanism for the export of cellular lipids in response to thermal stress.

Keywords: heat stress, granulosa cells, theca cells, steroidogenesis, chicken, apolipoprotein B, microsomal triglyceride transfer protein

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Nur Farah Ain Zainee, Ahmad Ismail, Nazlina Ibrahim, Asmida Ismail

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Seaweeds are economically important as they have the potential of being utilized, the capabilities and opportunities for further expansion as well as the availability of other species for future development. Hence, research on the diversity and distribution of seaweeds have to be expanded whilst the seaweeds are one of the Malaysian marine valuable heritage. The study on the distribution of seaweeds at Pengerang, Johor was carried out between February and November 2015 at Kampung Jawa Darat and Kampung Sungai Buntu. The study sites are located at the south-southeast of Peninsular Malaysia where the Petronas Refinery and Petrochemicals Integrated Project Development (RAPID) are in progress. In future, the richness of seaweeds in Pengerang will vanish soon due to the loss of habitat prior to RAPID project. The research was completed to study the diversity of seaweed and to determine the present of fungal endophyte isolated from the seaweed. The sample was calculated by using quadrat with 25-meter line transect by 3 replication for each site. The specimen were preserved, identified, processed in the laboratory and kept as herbarium specimen in Algae Herbarium, Universiti Kebangsaan Malaysia. The complete thallus specimens for fungal endophyte screening were chosen meticulously, transferred into sterile zip-lock plastic bag and kept in the freezer for further process. A total of 29 species has been identified including 12 species of Chlorophyta, 2 species of Phaeophyta and 14 species of Rhodophyta. From February to November 2015, the number of species highly varied and there was a significant change in community structure of seaweeds. Kampung Sungai Buntu shows the highest diversity throughout the study compared to Kampung Jawa Darat. This evidence can be related to the high habitat preference such as types of shores which is rocky, sandy and having lagoon and bay. These can enhance the existence of the seaweeds community due to variations of the habitat. Eighteen seaweed species were selected and screened for the capability presence of fungal endophyte; Sargassum polycystum marked having the highest number of fungal endophyte compared to the other species. These evidence has proved the seaweed have capable of accommodating a lot of species of fungal endophytes. Thus, these evidence leads to positive consequences where further research should be employed.

Keywords: diversity, fungal endophyte, macroalgae, screening, seaweed

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Authors: Bongmun Kang, Kagnari Yamakawa, Yoshihisa Hagihara, Yuji Ito, Michimasa Kishimoto, Yoichi Kumada

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The PMMA-tag was genetically fused with the C-terminal region of VHH molecules. This antibody, VHH, is known as a single-chain domain, which is devoid of light chains. The PMMA-tag, which could affect the isoelectric point (pI) changeable with a charge of amino acid in VHHs were closely related to the solubility of VHH molecules during refolding. The genetic fusion of PMMA-tag to C-terminal region of VHHs significantly affects the recovery of their soluble protein during refolding by 50 mM TAPS at pH 8.5. It could be refolded with a recovery of more than 95% by dialysis at pH 8.5. A marked difference in the antigen-binding activities in the adsorption state was significantly high in VHH-PM compared to the wild type of VHH. There are approximately 8-fold differences in the antigen-binding activities in the adsorption state between VHH-PM and VHH.

Keywords: VHH, PMMA-tag, isoelectric point, pH, Solubility, refolding, immobilization, ELISA

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Authors: Sudhir Kumar Sharma, Abiy D. Woldetsadik, Mazin Magzoub, Ramesh Jagannathan

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It is well known that bioactive ceramics exhibit specific biological affinities, especially in the area of tissue re-generation. In this context, we report the development of an eminently scalable, novel, supercritical CO₂ based process for the fabrication of hierarchically porous 'soft' CaCO₃ scaffolds. Porosity at the macro, micro, and nanoscales was obtained through process optimization of the so-called 'coffee-ring effect'. Exposure of these CaCO₃ scaffolds to monocytic THP-1 cells yielded negligible levels of tumor necrosis factor-alpha (TNF-α) thereby confirming the lack of immunogenicity of the scaffolds. ECM protein treatment of the scaffolds showed enhanced adsorption comparable to standard control such as glass. In vitro studies using osteoblast precursor cell line, MC3T3, also demonstrated the cytocompatibility of hierarchical porous CaCO₃ scaffolds.

Keywords: supercritical CO2, CaCO3 scaffolds, coffee-ring effect, ECM proteins

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Authors: Vasile Simulescu, Jakub Mondek, Miloslav Pekař

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Hyaluronic acid (HA) is an anionic glycosaminoglycan distributed throughout connective, epithelial and neural tissues. The importance of hyaluronic acid increased in the last decades. It has many applications in medicine and cosmetics. Hyaluronic acid has been used in attempts to treat osteoarthritis of the knee via injecting it into the joint. Bovine serum albumin (also known as BSA) is a protein derived from cows, which has many biochemical applications. The aim of our research work was to compare the thermal degradation of hyaluronic acid and BSA in powder and in solution, by determining changes in molar mass and conformation, by using SEC-MALLS (size exclusion chromatography -multi angle laser light scattering). The aim of our research work was to observe the degradation in powder and in solution of different molar mass hyaluronic acid samples, at different temperatures for certain periods. The degradation of the analyzed samples was mainly observed by modifications in molar mass.

Keywords: thermal degradation, hyaluronic acid, BSA, SEC-MALLS

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