Search results for: animal protein (AP)

Authors: Sulhee Lee, Geon Kim, Young-Seo Park

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Unlike the traditional milk sterilization methods (LTLT, HTST, or UHT), pulsed electric field (PEF) technology is a non-thermal pasteurization process. This technology minimizes energy required for heat treatment in food processing, changes in sensory properties, and physical losses. In this study, structural changes of bovine milk proteins, the amount of immunoproteins such as IgG, and their storability by PEF treatment were examined. When the changes of protein content in PEF-treated milk were examined using HPLC, the amounts of α-casein and β-lactoglobulin were reduced over 40% each, whereas those of κ-casein and β-casein did not change. The amount of α-casein in HTST milk was reduced to 50%. When PEF was applied to milk at the energy level of 250 kJ, the amounts of IgG, IgA, β-lactoglobulin (β-LG), lactoferrin, and α-lactalbumin (α-LA) decreased by 43, 41, 35, 63, and 45%, respectively. When milk was sterilized by LTLT process followed by PEF process at the level of 150 kJ, the concentrations of IgG, IgA, β-LG, lactoferrin, and α-LA were 56.6, 10.6, 554, 2.8 and 660.1 μg/mL, respectively. When the bovine milk was sterilized by LTLT process followed by PEF process at the energy level of 180 kJ, storability of immunoproteins of milk was the highest and the concentrations of IgG, IgA, and β-LG decreased by 79.5, 6.5, and 134.5 μg/mL, respectively, when compared with the initial concentrations of those proteins. When bovine milk was stored at 4℃ after sterilization through HTST sterilizer followed by PEF process at the energy level of 200 kJ, the amount of lactoferrin decreased 7.3% after 36 days of storage, whereas that of lactoferrin of raw milk decreased 16.4%. Our results showed that PEF treatment did not change the protein structure nor induce protein denaturation in milk significantly when compared with LTLT or HTST sterilization. Also, LTLT or HTST process in combination with PEF were more effective than LTLT only or HTST only process in the conservation of immunoproteins in bovine milk.

Keywords: pulsed electric field, bovine milk, immunoproteins, sterilization

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Authors: Bello Muhammad Dogon Kade

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The study was conducted to determine the effect of a treated groundnut (Arachis hypogaea) shell in a complete diet on blood metabolites and microbial load in fattening Yankasa rams. The study was conducted at the Teaching and Research Farm (Small Ruminants Unit of Animal Science Department, Faculty of Agriculture, Ahmadu Bello University, Zaria. Each kilogram of groundnut shell was treated with 5% urea and 5% lime for treatments 2 (UTGNS) and 3 (LTGNS), respectively. For treatment 4 (ULTGNS), 1 kg of groundnut shell was treated with 2.5% urea and 2.5% lime, but the shell in treatment 1 was not treated (UNTGNS). Sixteen Yankasa rams were used and randomly assigned to the four treatment diets with four animals per treatment in a completely randomized design (CRD). The diet was formulated to have 14% crude protein (CP) content. Rumen fluid was collected from each ram at the end of the experiment at 0 and 4 hours post-feeding. The samples were then put in a 30 ml bottle and acidified with 5 drops of concentrated sulphuric (0.1N H₂SO4) acid to trap ammonia. The results of the blood metabolites showed that the mean values of NH₃-N differed significantly (P<0.05) among the treatment groups, with rams in the ULTGNS diet having the highest significant value (31.96 mg/L). TVFs were significantly (P<0.05) higher in rams fed UNTGNS diet and higher in total nitrogen; the effect of sampling periods revealed that NH3N, TVFs and TP were significantly (P<0.05) higher in rumen fluid collected 4hrs post feeding among the rams across the treatment groups, but rumen fluid pH was significantly (p<0.05) higher in 0-hour post-feeding in all the rams in the treatment diets. In the treatment and sampling period’s interaction effects, animals on the ULTGNS diet had the highest mean values of NH3N in both 0 and 4 hours post-feeding and were significantly (P<0.5) higher compared to rams on the other treatment diets. Rams on the UTGNS diet had the highest bacteria load of 4.96X105/ml, which was significantly (P<0.05) higher than a microbial load of animals fed UNTGNS, LTGNS and ULTGNS diets. However, protozoa counts were significantly (P<0.05) higher in rams fed the UTGNS diet than those followed by the ULTGNS diet. The results showed that there was no significant difference (P>0.05) in the bacteria count of the animals at both 0 and 4 hours post-feeding. But rumen fungi and protozoa load at 0 hours were significantly (P<0.05) higher than at 4 hours post-feeding. The use of untreated ground groundnut shells in the diet of fattening Yankasa ram is therefore recommended.

Keywords: blood metabolites, microbial load, volatile fatty acid, ammonia, total protein

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

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Authors: Hainan Chen, Jina Qing, Xiao Zhu, Ling Gao, Ampadu O. Jackson, Min Zhang, Kai Yin

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Objective: Editing macrophage activation to dampen inflammatory diseases by promoting the repolarization of inflammatory (M1) macrophages to anti-inflammatory (M2) macrophages is highly associated with mitochondrial respiration. Recent studies have suggested that mitochondrial apolipoprotein A-1 binding protein (APOA1BP) was essential for the cellular metabolite NADHX repair to NADH, which is necessary for the mitochondrial function. The exact role of APOA1BP in the repolarization of M1 to M2, however, is uncertain. Material and method: THP-1-derived macrophages were incubated with LPS (10 ng/ml) or/and IL-4 (100 U/ml) for 24 hours. Biochemical parameters of oxidative phosphorylation and M1/M2 markers were analyzed after overexpression of APOA1BP in cells. Results: Compared with control and IL-4-exposed M2 cells, APOA1BP was downregulated in M1 macrophages. APOA1BP restored the decline in mitochondrial function to improve metabolic and phenotypic reprogramming of M1 to M2 macrophages. Blocking oxidative phosphorylation by oligomycin blunts the effects of APOA1BP on M1 to M2 repolarization. Mechanistically, LPS triggered the hydration of NADH and increased its hydrate NADHX which inhibit cellular NADH dehydrogenases, a key component of electron transport chain for oxidative phosphorylation. APOA1BP decreased the level of NADHX via converting R-NADHX to biologically useful S-NADHX. The mutant of APOA1BP aspartate188, the binding site of NADHX, fail to repair oxidative phosphorylation, thereby preventing repolarization. Conclusions: Restoring mitochondrial function by increasing mitochondrial APOA1BP might be useful to improve the reprogramming of inflammatory macrophages into anti-inflammatory cells to control inflammatory diseases.

Keywords: inflammatory diseases, macrophage repolarization, mitochondrial respiration, apolipoprotein A-1 binding protein, NADHX, NADH

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Authors: Radovan Cobanovic, Milica Rankov Sicar

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Biodiesel refers to a vegetable oil - or animal fat-based diesel fuel consisting of long-chain alkyl (methyl, ethyl, or propyl) esters. It is derived by alcoholysis of triacylglycerols (triglycerides) from various lipid based materials that can be traditionally categorized into the following main groups: vegetable oils, animal fats, waste and algal oils. The goal of this study was to evaluate microbiological stability of biodiesel samples since it has been made from vegetable oil or animal fat which was stored on room temperature. For the purposes of this study, analyzes were conducted on six samples of biodiesel first at zero sample at the reception day than fifth, thirtieth, sixtieth, ninetieth and one hundred twentieth day from the day of reception. During this period, biodiesel samples were subjected to microbiological analyses (Salmonella spp., Listeria monocytogenes, Enterobacteriaceae and total plate count). All analyses were tested according to ISO methodology: Salmonella spp ISO 6579, Listeria monocytogenes ISO 11290-2, Enterobacteriaceae ISO 21528-1, total plate count ISO 4833-1. The results obtained after the analyses which were done according to the plan during the 120 days indicate that are no changes of products concerning microbiological analyses. Salmonella spp., Listeria monocytogenes, Enterobacteriaceae were not detected and results for total plate count showed values < 10 cfu/g for all six samples. On the basis of this monitoring under defined storage conditions at room temperatures, the results showed that biodiesel is very stable as far as microbiological analysis were concerned.

Keywords: biodiesel, microbiology, room temperature, stability

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Authors: Hamid Mustafa, Adeela Ajmal, Kim EuiSoo, Noor-ul-Ain

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World wild, buffalo production is considered as most important component of food industry. Efficient buffalo production is related with reproductive performance of this species. Lack of knowledge of reproductive efficiency and its related genes in buffalo species is a major constraint for sustainable buffalo production. In this study, we performed some bioinformatics analysis on Follicle Stimulating Hormone Receptor (FSHR) gene and explored the possible relationship of this gene among different buffalo breeds and with other farm animals. We also found the evolution pattern for this gene among these species. We investigate CDS lengths, Stop codon variation, homology search, signal peptide, isoelectic point, tertiary structure, motifs and phylogenetic tree. The results of this study indicate 4 different motif in this gene, which are Activin-recp, GS motif, STYKc Protein kinase and transmembrane. The results also indicate that this gene has very close relationship with cattle, bison, sheep and goat. Multiple alignment (MA) showed high conservation of motif which indicates constancy of this gene during evolution. The results of this study can be used and applied for better understanding of this gene for better characterization of Follicle Stimulating Hormone Receptor (FSHR) gene structure in different farm animals, which would be helpful for efficient breeding plans for animal’s production.

Keywords: buffalo, FSHR gene, bioinformatics, production

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Authors: Samantha Shallop, Hakinya Karra, Tytus Bernas, Gladys Shaw, Gretchen Neigh, Jeffrey Dupree, Mathula Thangarajh

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Abnormal neuronal homeostasis is considered a structural correlate of cognitive deficits in Duchenne Muscular Dystrophy. Neurons are highly polarized cells with multiple dendrites but a single axon. Trafficking of cellular organelles are highly regulated, with the cargo in the somatodendritic region of the neuron not permitted to enter the axonal compartment. We investigated the molecular mechanisms that regular organelle trafficking in neurons using a multimodal approach, including high-resolution structural illumination, proteomics, immunohistochemistry, and computational modeling. We investigated the expression of ankyrin-G, the master regulator controlling neuronal polarity. The expression of ankyrin G and the morphology of the axon initial segment was profoundly abnormal in the CA1 hippocampal neurons in the mdx52 animal model of DMD. Ankyrin-G colocalized with kinesin KIF5a, the anterograde protein transporter, with higher levels in older mdx52 mice than younger mdx52 mice. These results suggest that the functional trafficking from the somatodendritic compartment is abnormal. Our data suggests that dystrophin deficiency compromised neuronal homeostasis via ankyrin-G-based mechanisms.

Keywords: neurons, axonal transport, duchenne muscular dystrophy, organelle transport

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Authors: Zeiler M., Stadler D., Schmaderer C.

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Over the years of dialysis treatment, most patients experience significant weight loss. The primary emphasis in earlier research was the underlying mechanism of protein energy wasting and the subsequent malnutrition inflammation syndrome. In the interest to provide an effective and rapid solution for the patients, the aim of this study is identifying individual influences of their assumed reduced dietary intake, such as nausea, appetite loss and taste changes, and to determine whether the patients adhere to their nutrition guidelines. A prospective, controlled study with 38 end-stage renal disease patients was performed using a questionnaire to reflect their diet within the last 12 months. Thereby, the daily intake for the most important macro-and micronutrients was calculated to be compared with the individual KDQOI-guideline value, as well as controls matched in age and gender. The majority of the study population did not report symptoms commonly associated with dialysis, such as nausea or inappetence, and denied any change in dietary behavior since receiving renal replacement therapy. The patients’ daily intake of energy (3080kcal ± 1266) and protein (89,9g [53,4-142,0]) did not differ significantly from the controls (energy intake: 3233kcal ± 1046, p=0,597; protein intake: 103,7g [90,1-125,5], p=0,120). The average difference to the individual calculated KDQOI-guideline was +176,0kcal ± 1156 (p=0,357) for energy intake and -1,75g ± 45,9 (p=0,491) for protein intake. However, there was an observed imbalance in the distribution of macronutrients, with a preference for fats over proteins. The patients’ daily intake of sodium (5,4g [ 2,95-10,1]) was higher than in the controls (4,1g [2,04-5,99], p= 0,058) whereas both values for potassium (3,7g ± 1,84) and phosphorous (1,79g ± 0,91) went significantly below the controls’ values (potassium intake: 4,89g ± 1,74, p=0,014; phosphorous intake: 2,04g ± 0,64, p=0,038). Thus, the values exceeded the calculated KDQOI-recommendation by + 3,3g [0,63-7,90] (p<0,001) for sodium, +1,49g ± 1,84 (p<0,001) for potassium and +0,89g ± 0,91 (p<0,001) for phosphorous. Contrary to the assumption, the patients did not under-eat. Nevertheless, their diets did not align with the recommended values. These findings highlight the need for intervention and education among patients and that regular dietary monitoring could prevent unhealthy nutrition habits. The elaboration of individual references instead of standardized guidelines could increase the compliance to the advised diet so that interdisciplinary comorbidities do not develop or worsen.

Keywords: compliance, dialysis, end-stage renal disease, KDQOI, malnutrition, nutrition guidelines, questionnaire, salt intake

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Authors: Ajith K. A. Gedara, Iva Chianella, Jose L. Endrino, Qi Zhang

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The olive seed is a by-product of the olive oil production industry. Biuret test and ferric chloride test revealed that water or alkali NaOH extractions of olive seed flour are rich in proteins and tannins. Both protein and tannins are well-known bio-based wood adhesives in the wood-based panel industry. In general, tannins-based adhesives show better mechanical and physical properties than protein wood adhesives. This paper explores different methods of extracting tannins from olive seed flour against the tannins yield and their applications as bio-based adhesives in wood-based panels. Once investigated, the physical and the mechanical properties of wood-based panels made using bio-adhesives based tannins extracted from olive seed flour revealed that the resulting products seemed to satisfy the Japanese Industrial Standards JIS A 5908:2015.

Keywords: bio-adhesives, olive seed flour, tannins, wood-based panels

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Authors: Maha Mohamed Hady Ali, M. A. El-Sayed

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Camel (Camelus dromedarius) is very important animal in many arid and semi-arid zones of tropical and subtropical regions as it serves as dual purpose providing meat and milk for human and as draft animal. Camel, like other animal must receive all essential nutrients despite the hostile environment. A study was conducted to evaluate the nutritional status of some micro-minerals of camel under Egyptian environmental condition. Forty five blood samples were collected from apparently healthy male camels with an average age between 2-6 years at the slaughter house in Cairo province, Egypt. The animals were fed mainly on berseem (Trifolium alexandrinum) or concentrate with straw before slaughtering. The collected serum and feedstuff samples were subjected to copper, iron, selenium and zinc analysis using Atomic absorption spectrophotometer. The data showed variation in the level of copper, iron, selenium and zinc in the serum of the dromedary camel as well as in the feedstuffs. Furthermore, the results indicated that the micro- minerals status of feeds may not always reflected as such in camel blood suggesting some role of bioavailability. The main reason for the lack of such reflection seems to be the wide diversity exists in the surrounding environment (forages and plants) as well as the bioavailability of such minerals. Since the requirement of micro-minerals have not been established for camel, more researches must be focused on this topic.

Keywords: camel, copper, egypt, feed stuff, iron, selenium, zinc

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Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

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Authors: Sarah Elsayed Mohammed, Lamiaa Ahmed Ahmed, Mahmoud Mohammed Khattab

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Aim: The aim of the present study was to investigate whether combined nicorandil and bone marrow-derived mesenchymal stem cells (BMDMSC) treatment could offer an additional benefit in ameliorating isoproterenol (ISO)-induced heart failure in rats. Methods: ISO (85 and 170 mg/kg/day) was injected subcutaneously for 2 successive days, respectively. By day 3, electrocardiographic changes were recorded and serum was separated for determination of CK-MB level for confirmation of myocardial damage. Nicorandil (3 mg/kg/day) was then given orally with or without a single i.v. BMDMSC administration. Electrocardiography and echocardiography were recorded 2 weeks after beginning of treatment. Rats were then sacrificed and ventricles were isolated for estimation of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) contents, caspase-3 activity as well as inducible nitric oxide synthase (iNOS) and connexin-43 protein expressions. Moreover, histological analysis of myocardial fibrosis was performed and cryosections were done for estimation of homing of BMDMSC. Results: ISO induced a significant increase in ventricles/body weight ratio, left ventricular end diastolic (LVEDD) and systolic dimensions (LVESD), ST segment and QRS duration. Moreover, myocardial fibrosis as well as VEGF, TNF-α and TGF-β contents were significantly increased. On the other hand, connexin-43 protein expression was significantly decreased, while caspase-3 and iNOS protein expressions were significantly increased. Combined therapy provided additional improvement compared to cell treatment alone towards reducing cardiac hypertrophy, fibrosis and inflammation. Furthermore, combined therapy induced significant increase in angiogenesis and BMDMSC homing and prevented ISO induced changes in iNOS, connexin-43 and caspase-3 protein expressions. Conclusion: Combined nicorandil/BMDMSC treatment was superior to BMDMSC alone towards preventing ISO-induced heart failure in rats.

Keywords: fibrosis, isoproterenol, mesenchymal stem cells, nicorandil

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Authors: Nurcan Cetinkaya, Nadide Hulya Ozdemir

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The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to the small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. . The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinin levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in µmol/W0. 75 (p<0.05). Mean creatinine values in µmol/W0. 75 and allantoin:creatinin ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage: concentrate ratio in Kıvırcık Yearlings.

Keywords: allantoin, creatinin, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisia

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Authors: Jill Steel

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Introduction: Animal-assisted learning (AAL) interventions are increasing exponentially, yet a paucity of quality research in the field exists. The aim of this study was to evaluate how the promotion of emotional wellbeing, through AAL, in this case, a dog, may support children’s engagement with reading in a Primary 1 classroom. Research indicates that dogs can provide emotional support to children; by forming a trusting attachment with a non-critical ‘friend’ who confers unconditional positive regard on the child, confidence may be boosted and anxiety reduced. By promoting emotional wellbeing through interactions with the dog, it is hoped that children begin to associate reading with feelings of wellbeing, which then results in increased engagement with reading. Methodology: A review of the literature was conducted. The relationship between emotional wellbeing and learning was explored, followed by an examination of the literature relating to Animal-Assisted Therapy and AAL. Scottish educational policy and legislation were analysed to establish the extent to which AAL might be suitable for the Scottish pedagogical context. An empirical study was conducted in a mainstream Primary 1 classroom over a four-week period. An inclusive approach was adopted whereby all children that wanted to interact with the dog were given the opportunity to do so, and all 25 children subsequently chose to participate. Children were not withdrawn from the classroom. Primary methods included interviews, observations, and questionnaires. Three focus children were selected for closer study. Main Results: Results were remarkably close to previous research and literature. Children’s emotional wellbeing was boosted, and engagement in reading improved. Principal Conclusions and Implications for Field: It was concluded that AAL could support emotional wellbeing and, in turn, promote children’s engagement with reading. The main limitation of the study was its short-term nature, and a longer randomised controlled trial with a larger sample, currently being undertaken by the author, would provide a fuller answer to the research question. Barriers to AAL include health and safety concerns and steps to ensure the welfare of the dog.

Keywords: animal-assisted learning, emotional wellbeing, reading, reading to dogs

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Authors: Sonaal Ramsookmohan

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Legume seeds are common foods in human diet and have been identied as a valuable source of human nutritonn Since they are useful sources of protein; legume proteins are used in many food applicatonsn Critcal functonal propertes are recognized to impact the quality of foodn Cowpea (Vigna unguiculata), has been well documented for its immense potental in contributng to food security forming part of daily staple diets in most developing countriesn. In this study, cowpea seeds were used to prepare cowpea four, protein isolates by the salt extractonndialysis method and peptdes by enzymatc hydrolysis using Alcalase and Flavourzymen Functonal analyses such as water absorpton capacity, oil absorpton capacity, emulsifying and foaming propertes were conducted on the cowpea peptdesn The physicochemical propertes determine their potental applicaton in food industries as functonal ingredientsn Cowpea peptdes could increase the value of cowpea by expanding its use, as well as contribute to the legume grain sector.

Keywords: physicochemical, peptides, Cowpea, alcalase, flavourzyme

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Authors: Rezvan Najafi, Korosh Heydari, Massoud Saidijam

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5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.

Keywords: colorectal cancer, miR-200c, 5-FU resistance, E-cadherin, PTEN

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Authors: Xi Chen, King-Yip Cheng

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Hypercholesterolemia, characterized by high low-density lipoprotein cholesterol (LDL-C), raises cardiovascular events in patients with type 2 diabetes (T2D). Although several drugs, such as statin and PCSK9 inhibitors, are available for the treatment of hypercholesterolemia, they exert detrimental effects on glucose metabolism and hence increase the risk of T2D. On the other hand, the drugs used to treat T2D have minimal effect on improving the lipid profile. Therefore, there is an urgent need to develop treatments that can simultaneously improve glucose and lipid homeostasis. Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 2 (APPL2) causes insulin resistance in the liver and skeletal muscle via inhibiting insulin and adiponectin actions in animal models. Single-nucleotide polymorphisms in the APPL2 gene were associated with LDL-C, non-alcoholic fatty liver disease, and coronary artery disease in humans. The aim of this project is to investigate whether APPL2 antisense oligonucleotide (ASO) can alleviate dietary-induced T2D and hypercholesterolemia. High-fat diet (HFD) was used to induce obesity and insulin resistance in mice. GalNAc-conjugated APPL2 ASO (GalNAc-APPL2-ASO) was used to silence hepatic APPL2 expression in C57/BL6J mice selectively. Glucose, lipid, and energy metabolism were monitored. Immunoblotting and quantitative PCR analysis showed that GalNAc-APPL2-ASO treatment selectively reduced APPL2 expression in the liver instead of other tissues, like adipose tissues, kidneys, muscle, and heart. The glucose tolerance test and insulin sensitivity test revealed that GalNAc-APPL2-ASO improved glucose tolerance and insulin sensitivity progressively. Blood chemistry analysis revealed that the mice treated with GalNAc-APPL2-ASO had significantly lower circulating levels of total cholesterol and LDL cholesterol. However, there was no difference in circulating levels of high-density lipoprotein (HDL) cholesterol, triglyceride, and free fatty acid between the mice treated with GalNac-APPL2-ASO and GalNAc-Control-ASO. No obvious effect on food intake, body weight, and liver injury markers after GalNAc-APPL2-ASO treatment was found, supporting its tolerability and safety. We showed that selectively silencing hepatic APPL2 alleviated insulin resistance and hypercholesterolemia and improved energy metabolism in the dietary-induced obese mouse model, indicating APPL2 as a therapeutic target for metabolic diseases.

Keywords: APPL2, antisense oligonucleotide, hypercholesterolemia, type 2 diabetes

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Authors: Zarmina Gillani, Mulazim Hussain Bukhari, Nuzhat Huma, Aqsa Qayyum

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Whey is a valuable by-product of dairy industry comprising of precious nutrients lactose, protein, vitamins and minerals for the human food but considered as a pollutant due to its biological activity. So, there is a need to develop nutritious whey products to overcome the problem of environmental pollution. This project was planned to develop a whey drink at different pasteurization temperatures and its quality was evaluated during storage. The result indicated that pH, acidity, total soluble solids and lactose content changed significantly (p < 0.01) due to lactic acid production during storage. Non-significant (p > 0.05) effects were detected on the protein and ash content of whey drink. Fat and viscosity changed significantly with respect to storage only. Sensory evaluation of whey drink revealed that both treatments remained acceptable while whey drink pasteurized at 75°C/30 minutes (WD2) gained more sensory score compared to whey drink pasteurized at 65°C/30minutes (WD1).

Keywords: pasteurization, sensory evaluation, storage, whey

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Authors: Viviane P. Romani, Bradley D. Olsen, Vilásia G. Martins

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Biodegradable films for food packaging have gained growing attention due to environmental pollution caused by synthetic films and the interest in the better use of resources from nature. Important research advances were made in the development of materials from proteins, polysaccharides, and lipids. However, the commercial use of these new generation of sustainable materials for food packaging is still limited due to their low mechanical and barrier properties that could compromise the food quality and safety. Thus, strategies to improve the performance of these materials have been tested, such as chemical modifications, incorporation of reinforcing structures and others. Cold plasma is a versatile, fast and environmentally friendly technology. It consists of a partially ionized gas containing free electrons, ions, and radicals and neutral particles able to react with polymers and start different reactions, leading to the polymer degradation, functionalization, etching and/or cross-linking. In the present study, biodegradable films from fish protein prepared through the casting technique were plasma treated using an AC glow discharge equipment. The reactor was preliminary evacuated to ~7 Pa and the films were exposed to air plasma for 2, 5 and 8 min. The films were evaluated by their mechanical and water vapor permeability (WVP) properties and changes in the protein structure were observed using Scanning Electron Microscopy (SEM) and X-ray diffraction (XRD). Potential cross-links and elimination of surface defects by etching might be the reason for the increase in tensile strength and decrease in the elongation at break observed. Among the times of plasma application tested, no differences were observed when higher times of exposure were used. The X-ray pattern showed a broad peak at 2θ = 19.51º that corresponds to the distance of 4.6Å by applying the Bragg’s law. This distance corresponds to the average backbone distance within the α-helix. Thus, the changes observed in the films might indicate that the helical configuration of fish protein was disturbed by plasma treatment. SEM images showed surface damage in the films with 5 and 8 min of plasma treatment, indicating that 2 min was the most adequate time of treatment. It was verified that plasma removes water from the films once weight loss of 4.45% was registered for films treated during 2 min. However, after 24 h in 50% of relative humidity, the water lost was recovered. WVP increased from 0.53 to 0.65 g.mm/h.m².kPa after plasma treatment during 2 min, that is desired for some foods applications which require water passage through the packaging. In general, the plasma technology affects the properties and structure of fish protein films. Since this technology changes the surface of polymers, these films might be used to develop multilayer materials, as well as to incorporate active substances in the surface to obtain active packaging.

Keywords: fish protein films, food packaging, improvement of properties, plasma treatment

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Authors: Keyur Raval, Steffen Krohn, Bruno Moerschbacher

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Chitin, the biopolymer of the N-acetylglucosamine, is the most abundant biopolymer on the planet after cellulose. Industrially, chitin is isolated and purified from the shell residues of shrimps. A deacetylated derivative of chitin i.e. chitosan has more market value and applications owing to it solubility and overall cationic charge compared to the parent polymer. This deacetylation on an industrial scale is performed chemically using alkalis like sodium hydroxide. This reaction not only is hazardous to the environment owing to negative impact on the marine ecosystem. A greener option to this process is the enzymatic process. In nature, the naïve chitin is converted to chitosan by chitin deacetylase (CDA). This enzymatic conversion on the industrial scale is however hampered by the crystallinity of chitin. Thus, this enzymatic action requires the substrate i.e. chitin to be soluble which is technically difficult and an energy consuming process. We in this project wanted to address this shortcoming of CDA. In lieu of this, we have modeled a fusion protein with CDA and an auxiliary protein. The main interest being to increase the accessibility of the enzyme towards crystalline chitin. A similar fusion work with chitinases had improved the catalytic ability towards insoluble chitin. In the first step, suitable partners were searched through the protein data bank (PDB) wherein the domain architecture were sought. The next step was to create the models of the fused product using various in silico techniques. The models were created by MODELLER and evaluated for properties such as the energy or the impairment of the binding sites. A fusion PCR has been designed based on the linker sequences generated by MODELLER and would be tested for its activity towards insoluble chitin.

Keywords: chitin deacetylase, modeling, chitin binding domain, chitinases

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Authors: Lali Shanshiashvili, Marika Chikviladze, Nino Mamulashvili, Maia Sepashvili, Nana Narmania, David Mikeladze

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Purpose: During demyelinating inflammatory diseases and after the damage of the myelin sheet, myelin-derived proteins, including myelin basic protein (MBP), are secreted into the extracellular space. MBP shows extensive post-translational modifications, including the deimination of arginine residues. Deiminated MBP is structurally less ordered, susceptible to proteolytic attack, and more immunogenic than the unmodified one. It is hypothesized that MBP could change the inflammatory response in astrocytes. Methods: MBP was isolated and purified from bovine brain white matter. Primary astrocyte cultures were prepared from whole brains of 2-day-old Wistar rats. For evaluation of glutamate uptake/release in astrocytes following treatment of cells with MBP charge isomers, Glutamate Assay Kit was used. The expression of EAAT-2 (excitatory amino acid transporters), peroxisome proliferator-activated receptor gamma (PPAR- γ), inhibitor of nuclear factor kappa B (IkB), and high mobility group protein B1 (HMGB1) in astrocytes were assayed by Western Blot analysis. Results: This study investigated the action of deiminated isomer (C8) on the cultured primary astrocytes and compared its effects with the effects of unmodified C1 isomers. The study found that C8 and C1 MBP differently act on the uptake and release of glutamate in astrocytes: nonmodified C1 MBP increases the uptake of glutamate and does not change the release, whereas C8 decreases the release of glutamate but does not alter the uptake. Nevertheless, both isomers increased the expression of PPAR-γ and EAAT2 in the same intensity. However, immunostaining and Western Blots of cell lysates showed a decrease of IkB and increased expression of HMGB1 after the treatment of astrocytes by C8. Moreover, in the presence of C8, astrocytes release more nitric oxide than unmodified C1 isomers. Conclusion: These data suggest that the deiminated isomer of MBP evokes an inflammatory response and enhances the ability of astrocytes to release proinflammatory mediators through activation of NF-kB after the breakdown of myelin sheets. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.

Keywords: myelin basic protein, glutamate, deimination, astrocytes, inflammation

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Authors: Satoshi Furukawa, Satomu Morita, Lisa Wingenfeld, Katsuji Nishi, Masahito Hitosugi

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We studied the expression of hypoxia-related antigens (e.g., cold-inducible antigens and apoptotic antigens) in the myocardium and the cerebellumthat were obtained from individuals after death by carbon monoxide or hypothermia. The immunohistochemistry results revealed that expression of cold-inducible RNA binding protein (CIRBP) and RNA-binding protein 3 (RBM3) may be associated with hpyothermic and the hypoxic conditions. The expression of CIRBP and RBM3 in the myocardium was different from their expression in the cerebellum, especially in the Purkinje cells. The results indicate that agonal duration influences antigen expression. In the hypothermic condition, the myocardium uses more ATP since the force of the excitation-contraction coupling of the myocardium increases by more than 400% when the experimental temperature is reduced from 35°C to 25°C. The results obtained in this study indicate that physicians should pay attention to the myocardium when cooling the patient’s body to protect the brain.

Keywords: carbon monoxide death, cerebellum, CIRBP, hypothermic death, myocardium, RBM3

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Authors: Azwan Hamdan, Mohd Qayyum Ab Latip, Hasliza Abu Hassim, Tengku Rinalfi Putra Tengku Azizan, Hafandi Ahmad

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Animal cognition provides some evidence for self-recognition, which is described as the ability to recognize oneself as an individual separate from the environment and other individuals. The mirror self-recognition (MSR) or mark test is a behavioral technique to determine whether an animal have the ability of self-recognition or self-awareness in front of the mirror. It also describes the capability for an animal to be aware of and make judgments about its new environment. Thus, the objectives of this study are to measure and to compare the ability of wild and captive wildlife in mirror self-recognition. Wild animals from the Royal Belum Rainforest Malaysia were identified based on the animal trails and salt lick grounds. Acrylic mirrors with wood frame (200 x 250cm) were located near to animal trails. Camera traps (Bushnell, UK) with motion-detection infrared sensor are placed near the animal trails or hiding spot. For captive wildlife, animals such as Malayan sun bear (Helarctos malayanus) and chimpanzee (Pan troglodytes) were selected from Zoo Negara Malaysia. The captive animals were also marked using odorless and non-toxic white paint on its forehead. An acrylic mirror with wood frame (200 x 250cm) and a video camera were placed near the cage. The behavioral data were analyzed using ethogram and classified through four stages of MSR; social responses, physical inspection, repetitive mirror-testing behavior and realization of seeing themselves. Results showed that wild animals such as barking deer (Muntiacus muntjak) and long-tailed macaque (Macaca fascicularis) increased their physical inspection (e.g inspecting the reflected image) and repetitive mirror-testing behavior (e.g rhythmic head and leg movement). This would suggest that the ability to use a mirror is most likely related to learning process and cognitive evolution in wild animals. However, the sun bear’s behaviors were inconsistent and did not clearly undergo four stages of MSR. This result suggests that when keeping Malayan sun bear in captivity, it may promote communication and familiarity between conspecific. Interestingly, chimp has positive social response (e.g manipulating lips) and physical inspection (e.g using hand to inspect part of the face) when they facing a mirror. However, both animals did not show any sign towards the mark due to lost of interest in the mark and realization that the mark is inconsequential. Overall, the results suggest that the capacity for MSR is the beginning of a developmental process of self-awareness and mental state attribution. In addition, our findings show that self-recognition may be based on different complex neurological and level of encephalization in animals. Thus, research on self-recognition in animals will have profound implications in understanding the cognitive ability of an animal as an effort to help animals, such as enhanced management, design of captive individuals’ enclosures and exhibits, and in programs to re-establish populations of endangered or threatened species.

Keywords: mirror self-recognition (MSR), self-recognition, self-awareness, wildlife

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Authors: Amena Omer Syeda, Harita Shyam

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Background: Anorexia a common symptom among patients with prolonged illness leading to anorexia-cachexia syndrome with a prevalence rate of 70%. In order to provide effective health care and better response to treatment, appetite should be assessed on admission and then periodically for earlier nutrition intervention. Functional Assessment of Anorexia/Cachexia Therapy (FAACT) appetite scale is 12 questions, patient-rated, symptom specific measure for appetite, and distress from anorexia. It assigns a score ranging from 0 (worst response) to 4 (best response). Therefore, proposing a total score of ≤24 may be sufficient to make a diagnosis of anorexia. Objectives: To assess the FAACT scale by co-relating the scores with the Nutritional intake and BMI of Cancer Patients. Methods: The FAACT scores of 100 cancer in-patients receiving chemotherapy or radiation as treatment, their 24-hour calorie and protein intake and BMI were recorded. The data was then statistically analyzed. Results: The calorie and protein intake and FAACT scores both showed a significant positive co-relation (p<0.001), inferring that the patients with a FAACT score of ≤24 where not meeting their calorie as well as protein requirements, hence rightly categorizing them as anorexic. The co-relation between BMI and FAACT scores showed a weak co-relation and was not statistically significant (p > 0.05).The FAACT scale thus is not sensitive to distinguish patients being under-weight, normal weight or obese. Conclusion: The FAACT scale helps in providing better palliative and nutritional care as it correctly assessed anorexia /cachexia in cancer patients and co-related significantly with their nutrient intake.

Keywords: appetite, cachexia, cancer, malnutrition

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Authors: Wenfa Yu, Thuva Gnanasampanthan, John Finlay, Jessica Clarke, Charlotte Anderson, Tony Clare, Axel Rosenhahn

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Marine biofouling is a worldwide problem at vast economic and ecological costs. Historically it was combated with toxic coatings such as tributyltin. As those coatings being banned nowadays, finding environmental friendly antifouling solution has become an urgent topic. In this study antifouling coatings consisted of natural occurring polysaccharides hyaluronic acid (HA), alginic acid (AA), chitosan (Ch) and polyelectrolyte polyethylenimine (PEI) are constructed into polyelectrolyte multilayers (PEMs) in a Layer-by-Layer (LbL) method. LbL PEM construction is a straightforward way to assemble biomacromolecular coatings on surfaces. Advantages about PEM include ease of handling, highly diverse PEM composition, precise control over the thickness and so on. PEMs have been widely employed in medical application and there are numerous studies regarding their protein adsorption, elasticity and cell adhesive properties. With the adjustment of coating composition, termination layer charge, coating morphology and cross-linking method, it is possible to prepare low marine biofouling coatings with PEMs. In this study, using spin coating technology, PEM construction was achieved at smooth multilayers with roughness as low as 2nm rms and highly reproducible thickness around 50nm. To obtain stability in sea water, the multilayers were covalently cross-linked either thermally or chemically. The cross-linking method affected surface energy, which was reflected in water contact angle, thermal cross-linking led to hydrophobic surfaces and chemical cross-linking generated hydrophilic surfaces. The coatings were then evaluated regarding its protein resistance and biological species resistance. While the hydrophobic thermally cross-linked PEM had low resistance towards proteins, the resistance of chemically cross-linked PEM strongly depended on the PEM termination layer and the charge of the protein, opposite charge caused high adsorption and same charge low adsorption, indicating electrostatic interaction plays a crucial role in the protein adsorption processes. Ulva linza was chosen as the biological species for antifouling performance evaluation. Despite of the poor resistance towards protein adsorption, thermally cross-linked PEM showed good resistance against Ulva spores settlement, the chemically cross-linked multilayers showed poor resistance regardless of the termination layer. Marine species adhesion is a complex process, although it involves proteins as bioadhesives, protein resistance its own is not a fully indicator for its antifouling performance. The species will pre select the surface, responding to cues like surface energy, chemistry, or charge and so on. Thus making it difficult for one single factors to determine its antifouling performance. Preparing PEM coating is a comprehensive work involving choosing polyelectrolyte combination, determining termination layer and the method for cross-linking. These decisions will affect PEM properties such as surface energy, charge, which is crucial, since biofouling is a process responding to surface properties in a highly sensitive and dynamic way.

Keywords: hyaluronic acid, polyelectrolyte multilayers, protein resistance, Ulva linza zoospores

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Authors: Chang Liang, Weizhi Gong, Yan Zhang

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Purpose: Hepatocellular carcinoma (HCC) is a malignant tumor with a high mortality rate and poor prognosis worldwide. Murine double minute 2 (MDM2) regulates the tumor suppressor p53, increasing cancer risk and accelerating tumor progression. Speckle-type POX virus and zinc finger protein (SPOP), a key of subunit of Cullin-Ring E3 ligase, inhibits tumor genesis and progression by the ubiquitination of its downstream substrates. This study aimed to clarify whether SPOP and MDM2 are mutually regulated in HCC and the correlation between SPOP and MDM2 and the prognosis of HCC patients. Methods: First, the expression of SPOP and MDM2 in HCC tissues were detected by TCGA database. Then, 53 paired samples of HCC tumor and adjacent tissues were collected to evaluate the expression of SPOP and MDM2 using immunohistochemistry. Chi-square test or Fisher’s exact test were used to analyze the relationship between clinicopathological features and the expression levels of SPOP and MDM2. In addition, Kaplan‒Meier curve analysis and log-rank test were used to investigate the effects of SPOP and MDM2 on the survival of HCC patients. Last, the Multivariate Cox proportional risk regression model analyzed whether the different expression levels of SPOP and MDM2 were independent risk factors for the prognosis of HCC patients. Results: Bioinformatics analysis revealed the low expression of SPOP and high expression of MDM2 were related to worse prognosis of HCC patients. The relationship between the expression of SPOP and MDM2 and tumor stem-like features showed an opposite trend. The immunohistochemistry showed the expression of SPOP protein was significantly downregulated while MDM2 protein significantly upregulated in HCC tissue compared to that in para-cancerous tissue. Tumors with low SPOP expression were related to worse T stage and Barcelona Clinic Liver Cancer (BCLC) stage, but tumors with high MDM2 expression were related to worse T stage, M stage, and BCLC stage. Kaplan–Meier curves showed HCC patients with high SPOP expression and low MDM2 expression had better survival than those with low SPOP expression and high MDM2 expression (P < 0.05). A multivariate Cox proportional risk regression model confirmed that a high MDM2 expression level was an independent risk factor for poor prognosis in HCC patients (P <0.05). Conclusion: The expression of SPOP protein was significantly downregulated, while the expression of MDM2 significantly upregulated in HCC. The low expression of SPOP and high expression. of MDM2 were associated with malignant progression and poor prognosis of HCC patients, indicating a potential therapeutic target for HCC patients.

Keywords: hepatocellular carcinoma, murine double minute 2, speckle-type POX virus and zinc finger protein, ubiquitination

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Authors: Fei Xu, Guofan Zhang, Xiao Liu

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Many major marine invertebrate phyla are characterized by indirect development. These animals transit from planktonic larvae to benthic adults via settlement and metamorphosis, which has many advantages for organisms to adapt marine environment. Studying the biological process of metamorphosis is thus a key to understand the origin and evolution of indirect development. Although the mechanism of metamorphosis has been largely studied on their relationships with the marine environment, microorganisms, as well as the neurohormones, little is known on the gene regulation network (GRN) during metamorphosis. We treated competent oyster pediveligers with epinephrine, which was known to be able to effectively induce oyster metamorphosis, and analyzed the dynamics of gene and proteins with transcriptomics and proteomics methods. The result indicated significant upregulation of protein synthesis system, as well as some transcription factors including Homeobox, basic helix-loop-helix, and nuclear receptors. The result suggested the GRN complexity of the transition stage during oyster metamorphosis.

Keywords: indirect development, gene regulation network, protein synthesis, transcription factors

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Authors: Padmanabhan D, Kavitha S

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The purpose of the study is to investigate arsenic resistance ability of indigenous microflora and its ability to utilize arsenic species form containing water source. PS-6 potential arsenic tolerance bacterium was screened from thirty isolates from Pichavaram Mangroves of India having tolerance to grow up to 1000 mg/l of As (V) and 800 mg/l of As (III) and arsenic utilization ability of 98 % of As (V) and 97% of As (III) with initial concentration of 3-5 mg/l within 48 hrs. Optimum pH and temperature was found to be ~7-7.4 and 37°C. Active growth of PS-6 in minimal salt media (MSB) helps in cost effective biomass production. Dry weight analysis of PS-6 has shown significant difference in biomass when exposed to As (III) and As (V). Protein level study of PS-6 after exposing to As (V) and As (III) shown modification in total protein concentration and variation in SDS-PAGE pattern. PS-6 was identified as Bacillus licheniformis based on partially sequenced of 16S rRNA using NCBI Blast. Further investigation will help in using this potential bacterium as a well-grounded source for urgency.

Keywords: arsenite, arsenate, Bacillus licheniformis, utilization

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Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

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Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

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Authors: Julie A. Carberry, Victor Maddalena

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Animal-Assisted Interventions (AAIs) have existed in various forms for centuries. In the past 30 years, there has been a dramatic increase in popularity. AAIs are now part of the lives of persons of all ages in many types of institutions. Anecdotal evidence of the benefits of AAIs have led to widespread adoption, yet there remains a lack of solid research base for support. The research question was, what are the lived experiences of AAI volunteer handlers are? An interpretive phenomenological methodology was used for this qualitative study. Data were collected from 1 - 2 hour-long semi-structured interviews and 1 observational field visit. All interviews were conducted, transcribed, and coded for themes by the principal investigator. Participants must have been an active St. John Ambulance Therapy Dog Program volunteer for a least one year. In total, 14 volunteer handlers, along with some of their dogs, were included. The St. John Ambulance is a not for profit organization that provides training and community services to Canadians. The Therapy Dog Program is 1 of the 4 nationally recognized core community service programs. The program incorporates dogs in the otherwise traditional therapeutic intervention of friendly visitation with clients. The lack of formal objectives and goals, and a trained therapist defines the program as an Animal-Assisted Activity (AAA), which is a type of AAI. Since the animals incorporated are dogs, the program is specifically a Canine-Assisted Activity (CAA), which is a type of Canine-Assisted Intervention (CAI). Six themes emerged from the analysis of the data: (a) a win-win-win situation for all parties involved – volunteer handlers, clients, and the dogs, (b) being on the other end of the leash: functions of the role of volunteer handler, (c) the importance of socialization: from spreading smiles to creating meaningful connections, (d) the role of the dog: initiating interaction and providing comfort, (e) an opportunity to feel good and destress, and (f) altruism versus personal rewards. Other insights were found regarding the program, clients, and staff. Possible implications from this research include increased organizational recruitment and retention of volunteer handlers and as well as increased support for CAAs and other CAIs that incorporate teams of volunteer handlers and their dogs. This support could, in turn, add overall support for the acceptance and broad implementation of AAIs as an alternative and or complementary non-pharmaceutical therapeutic intervention.

Keywords: animal-assisted activity, animal-assisted intervention, canine-assisted activity, canine-assisted intervention, perspective, qualitative, volunteer handler

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