Search results for: whey protein isolated
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3772

Search results for: whey protein isolated

922 Protective Effect of Saponin Extract from the Root of Garcinia kola (Bitter Kola) against Paracetamol-Induced Hepatotoxicity in Albino Rats

Authors: Alli Smith Yemisi Rufina, Adanlawo Isaac Gbadura

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Liver disorders are one of the major problems of the world. Despite its frequent occurrence, high morbidity, and high mortality, its medical management is currently inadequate. This study was designed to evaluate the Hepatoprotective effect of saponin extract of the root of Garcinia kola on the integrity of the liver of paracetamol induced Wistar albino rats. Twenty-five male adult Wistar albino rats were divided into five (5) groups. Group I, was the Control group that received distilled water only, group II was the negative control that received 2 g/kg of paracetamol on the 13th day, and group III, IV, and V were pre-treated with 100, 200 and 400 mg/kg of the saponin extract before inducing the liver damage on the 13th day with 2 g/kg of paracetamol. Twenty-four hours after administration, the rats were sacrificed, and blood samples were collected. The serum Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP) activities, Bilirubin and Conjugated Bilirubin, Glucose and Protein concentrations were evaluated. The liver was fixed immediately in Formalin and was processed and stained with Haematoxylin and Eosin (H&E). Administration of saponin extract from the root of Garcinia kola significantly decreased paracetamol induced elevated enzymes in the test group. Also, histological observations showed that saponin extract of the root of Garcinia kola exhibited a significant liver protection against the toxicant as evident by the cells trying to return to normal. Saponin extract from the root of Garcinia kola indicated a protection of the structural integrity of the hepatocytic cell membrane and regeneration of the damaged liver.

Keywords: hepatoprotective, liver damage, Garcinia kola, saponin, paracetamol

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921 Comparison Study of 70% Ethanol Effect on Direct and Retrival Culture of Contaminated Umblical Cord Tissue for Expansion of Mesenchymal Stem Cells

Authors: Ganeshkumar, Ashika, Valavan, Ramesh, Thangam, Chirayu

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MSCs are found in much higher concentration in the Wharton’s jelly compared to the umbilical cord blood, which is a rich source of hematopoietic stem cells. Umbilical cord tissue is collected at the time of birth; it is processed and stored in liquid nitrogen for future therapeutical purpose. The source of contamination might be either from vaginal tract of mother or from hospital environment or from personal handling during cord tissue sample collection. If the sample were contaminated, decontamination procedure will be done with 70% ethanol (1 minute) in order to avoid sample rejection. Ethanol is effective against a wide range of bacteria, protozoa and fungi and has low toxicity to humans. Among the 1954 samples taken for the study, 24 samples were found to be contaminated with microorganism. The organisms isolated from the positive samples were found to be E. coli, Stenotrophomonas maltophilia, Pseudomonas aueroginosa, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, Enterobacter cloacae, and Proteus mirabilis. Among these organisms 70% ethanol successfully eliminated E. coli, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, and Proteus mirabilis. 70% ethanol was unsuccessful in eliminating Stenotrophomonas maltophilia, Pseudomonas aueroginosa, and Enterobacter cloacae. Stenotrophomonas maltophilia and Pseudomonas aueroginosa have the ability to form biofilm that make them resistant to alcohol. Biofilm act as protective layer for bacteria and which protects them from host defense and antibiotic wash. Finally it was found 70% ethanol wash saved 58.3% cord tissue samples from rejection and it is ineffective against 41% of the samples. The contamination rate can be reduced by maintaining proper aseptic techniques during sample collection and processing.

Keywords: umblical cord tissue, decontamination, 70% ethanol effectiveness, contamination

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920 Localisation of Fluorescently Labelled Drug-Free Phospholipid Vesicles to the Cartilage Surface of Rat Synovial Joints

Authors: Sam Yurdakul, Nick Baverstock, Jim Mills

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TDT 064 (FLEXISEQ®) is a drug-free gel used to treat osteoarthritis (OA)-associated pain and joint stiffness. It contains ultra-deformable phospholipid Sequessome™ vesicles, which can pass through the skin barrier intact. In six randomized OA studies, topical TDT 064 was well tolerated and improved joint pain, physical function and stiffness. In the largest study, these TDT 064-mediated effects were statistically significantly greater than oral placebo and equivalent to celecoxib. To understand the therapeutic effects of TDT 064, we investigated the localisation of the drug-free vesicles within rat synovial joints. TDT 064 containing DiO-labelled Sequessome™ vesicles was applied to the knees of four 6-week-old CD® hairless rats (10 mg/kg/ joint), 2–3 times/day, for 3 days (representing the recommended clinical dose). Eighteen hours later, the animals and one untreated control were sacrificed, and the knee joints isolated, flash frozen and embedded in Acrytol Mounting Media™. Approximately 15 sections (10 µm) from each joint were analysed by fluorescence microscopy. To investigate whether the localisation of DiO fluorescence was associated with intact vesicles, an anti-PEG monoclonal antibody (mAb) was used to detect Tween, a constituent of Sequessome™ vesicles. Sections were visualized at 484 nm (DiO) and 647 nm (anti-PEG mAb) and analysed using inForm 1.4 (Perkin Elmer, Inc.). Significant fluorescence was observed at 484 nm in sections from TDT 064-treated animals. No non-specific fluorescence was observed in control sections. Fluorescence was detected as discrete vesicles on the cartilage surfaces, inside the cartilaginous matrix and within the synovial space. The number of DiO-labelled vesicles in multiple fields of view was consistent and >100 in sections from four different treated knees. DiO and anti-PEG mAb co-localised within the collagenous tissues in four different joint sections. Under higher magnification (40x), vesicles were seen in the intercellular spaces of the synovial joint tissue, but no fluorescence was seen inside cells. These data suggest that the phospholipid vesicles in TDT 064 localize at the surface of the joint cartilage; these vesicles may therefore be supplementing the phospholipid deficiency reported in OA and acting as a biolubricant within the synovial joint.

Keywords: joint pain, osteoarthritis, phospholipid vesicles, TDT 064

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919 The Effect of Low and High Dose Curcumin Supplementation on Prevention and Treatment of Sarcopenia: The Concept of Hormesis

Authors: Sevana Daneghian, Leila Chodari, Sahar Mehranfar, Shadi Mohammadpour-Asl, Diman Mahdi

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Sarcopenia is an age-related muscle disease. Lack of antioxidant protection, and cumulative oxidative damage to skeletal muscle are recognized mechanisms. Curcumin is a hormetin as it has a stimulating effect in low doses and an inhibitory effect in high doses. The purpose of this study was to examine the effects of four weeks of curcumin supplementation in presarcopenic and sarcopenic rats, and likelihood of potential negative effects while co-exist with sarcopenia. The rats were divided into 7 groups: young sham rats, 18 months old; presarcopenic control, supplemented with 400 and 1500 mg/kg/day, 24 months old; Sarcopenia control, sarcopenia supplemented with 400 and 1500 mg/kg/day. MDA levels were significantly reduced in the low-dose pre-sarcopenic group compared to the control group. Compared to the corresponding control groups, SOD levels decreased in the groups treated with low-dose presarcopenic supplementation and increased in the high-dose sarcopenic supplemented. GPx levels increased at both doses only in the sarcopenic group compared to the control group. SIRT-1 only increased at low doses in the sarcopenic groups and PGC-1α in both pre-sarcopenia groups compared to the corresponding control. IGF-1 increased compared to the control group at both doses in the pre-sarcopenic group and at high doses in sarcopenic group. Considering the hormetic effects of curcumin, it can be argued that, curcumin supplementation has some positive effects not only at low but also at high doses in both groups. This means that the high doses of curcumin have no negative synergistic effects in addition to oxidative stress during sarcopenia and high-dose supplementation in patients already suffering from high oxidative stress due to sarcopenia is safe and could be considered hormetic.

Keywords: curcumin, hormesis, sarcopenia, muscular atrophy, PGC protein, Sirtuins

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918 Genomic Surveillance of Bacillus Anthracis in South Africa Revealed a Unique Genetic Cluster of B- Clade Strains

Authors: Kgaugelo Lekota, Ayesha Hassim, Henriette Van Heerden

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Bacillus anthracis is the causative agent of anthrax that is composed of three genetic groups, namely A, B, and C. Clade-A is distributed world-wide, while sub-clades B has been identified in Kruger National Park (KNP), South Africa. KNP is one of the endemic anthrax regions in South Africa with distinctive genetic diversity. Genomic surveillance of KNP B. anthracis strains was employed on the historical culture collection isolates (n=67) dated from the 1990’s to 2015 using a whole genome sequencing approach. Whole genome single nucleotide polymorphism (SNPs) and pan-genomics analysis were used to define the B. anthracis genetic population structure. This study showed that KNP has heterologous B. anthracis strains grouping in the A-clade with more prominent ABr.005/006 (Ancient A) SNP lineage. The 2012 and 2015 anthrax isolates are dispersed amongst minor sub-clades that prevail in non-stabilized genetic evolution strains. This was augmented with non-parsimony informative SNPs of the B. anthracis strains across minor sub-clades of the Ancient A clade. Pan-genomics of B. anthracis showed a clear distinction between A and B-clade genomes with 11 374 predicted clusters of protein coding genes. Unique accessory genes of B-clade genomes that included biosynthetic cell wall genes and multidrug resistant of Fosfomycin. South Africa consists of diverse B. anthracis strains with unique defined SNPs. The sequenced B. anthracis strains in this study will serve as a means to further trace the dissemination of B. anthracis outbreaks globally and especially in South Africa.

Keywords: bacillus anthracis, whole genome single nucleotide polymorphisms, pangenomics, kruger national park

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917 The Role of Leukocyte-Derived IL-10 on Postoperative ileus and Intestinal Macrophage Differentiation in Mice

Authors: Kathy Stein, Mariola Lysson, Anja Schmidt, Beatrix Schumak, Sabine Specht, Hicham Bouabe, Jürgen Heesemann, Axel Roers, Joerg C. Kalff, Sven Wehner

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Objective: Postoperative ileus (POI) is a common complication of abdominal surgery. Monocyte infiltration is a hallmark of POI. The polarization of macrophages/monocytes in this process is not well understood. We aimed to investigate if and how M2 macrophage/monocyte differentiation is involved in POI pathogenesis. Design: POI was induced by intestinal manipulation (IM). C57Bl/6, CCR2-/-, IL-10 reporter (ITIB), IL-10-/- and LysMcre/IL-10fl/fl mice underwent IM. At various points in time leukocyte influx, gene and protein expression of cytokines, chemokines and M2 differentiation markers and intestinal motility were analyzed. Results: IM induced the postoperative expression of the M2 markers Arginase-1 and YM-1, predominantly in F4/80+Ly6C+ monocytes. Gene expression analyses indicated an IL-10-dependent, IL-4-independent M2 polarization of these monocytes. IL-10 dependency of M2 differentiation was confirmed in IL-10 deficient mice. Leukocytes, in the order of infiltrating monocytes, neutrophils, and resident macrophages were the main IL-10 producers during POI. IL-10 producing monocytes as well as M2 marker expression were almost absent in CCR2-deficient mice. However, postoperative IL-10 expression was not altered in CCR2-/- mice. The loss of M2 polarized monocytes neither protected CCR2-/- mice from nor affected resolution of POI. In contrast, IL-10 deficiency reduced postoperative neutrophil numbers and ameliorated POI. IL-10Ra expression was strongly induced in neutrophils but not in monocytes. Conclusion: We conclude that IL-10 counteracts POI resolution by activating IL-10Ra-expressing neutrophils in the late phase of disease while IL-10-dependent M2 differentiation is not pivotal to POI manifestation and resolution.

Keywords: interleukin-10, macrophages, neutrophils, postoperative ileus

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916 A Systematic Review with Meta-Analyses Investigating the Association between Binge Eating and Poor Weight Loss Outcomes in People with Obesity

Authors: Isabella Lobo Sasaoka, Felipe Q. da Luz, Zubeyir Salis, Phillipa Hay, Tamiris Gaeta, Paula Costa Teixeira, Táki Cordás, Amanda Sainsbury

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Background: A significant number of people with obesity that seek weight loss treatments experience binge eating episodes. Nonetheless, it is unknown whether binge eating episodes can hinder weight loss outcomes. Objective: To compare weight change in people with or without binge eating submitted to bariatric surgery, pharmacotherapy, nutritional orientation, and/or psychological therapies. Method: We conducted a systematic review with meta-analyses by searching studies in PubMed, American Psychological Association (APA), and Embase. Results: Thirty-four studies were included in our systematic review, and 17 studies were included in the meta-analyses. Overall, we found no significant difference in weight loss between people with or without binge eating submitted to any type of weight loss treatment. Additionally, we found no statistically significant differences in body weight between people with or without binge eating at short and long follow-up assessments following any type of weight loss treatment. We also examined changes in body weight in people with or without binge eating in three additional meta-analyses categorized by the type of weight loss treatment (i.e., behavioural and/or nutritional interventions; bariatric surgery; pharmacotherapy isolated or combined with behavior interventions) and found no difference in weight loss. Eleven out of the 17 studies that were assessed qualitatively (i.e., not included in meta-analyses) did not show differences in weight loss in people with or without binge eating submitted to any type of weight loss treatment. Conclusion: This systematic review with meta-analyses showed no difference in weight loss in people with or without binge eating submitted to a variety of weight loss treatments. Nonetheless, specialized therapies can be required to address eating disorder psychopathology and recurrent binge eating in people with obesity that seek weight loss.

Keywords: obesity, binge eating, weight loss, systematic review, meta-analysis

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915 Comparative Parametric Analysis on the Dynamic Response of Fibre Composite Beams with Debonding

Authors: Indunil Jayatilake, Warna Karunasena

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Fiber Reinforced Polymer (FRP) composites enjoy an array of applications ranging from aerospace, marine and military to automobile, recreational and civil industry due to their outstanding properties. A structural glass fiber reinforced polymer (GFRP) composite sandwich panel made from E-glass fiber skin and a modified phenolic core has been manufactured in Australia for civil engineering applications. One of the major mechanisms of damage in FRP composites is skin-core debonding. The presence of debonding is of great concern not only because it severely affects the strength but also it modifies the dynamic characteristics of the structure, including natural frequency and vibration modes. This paper deals with the investigation of the dynamic characteristics of a GFRP beam with single and multiple debonding by finite element based numerical simulations and analyses using the STRAND7 finite element (FE) software package. Three-dimensional computer models have been developed and numerical simulations were done to assess the dynamic behavior. The FE model developed has been validated with published experimental, analytical and numerical results for fully bonded as well as debonded beams. A comparative analysis is carried out based on a comprehensive parametric investigation. It is observed that the reduction in natural frequency is more affected by single debonding than the equally sized multiple debonding regions located symmetrically to the single debonding position. Thus it is revealed that a large single debonding area leads to more damage in terms of natural frequency reduction than isolated small debonding zones of equivalent area, appearing in the GFRP beam. Furthermore, the extents of natural frequency shifts seem mode-dependent and do not seem to have a monotonous trend of increasing with the mode numbers.

Keywords: debonding, dynamic response, finite element modelling, novel FRP beams

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914 Studies on Knockdown Resistance Mutations in Aedes aegypti and Aedes albopictus in India

Authors: Neera Kapoor

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Background: Knockdown Resistance (KDR) is one of the mechanisms of insecticide resistance in insects caused by the reduced target site sensitivity i.e. voltage gated sodium channel (VGSC) rendering it less sensitive to the toxic effects of DDT and pyrethroids. In this study, we evaluated insecticide susceptibility and its underlying KDR mechanism in eight Ae. aegypti and five Ae. albopictus field populations. Methodology: Field population was collected from four different geographical regions of India covering 18 districts of ten states. For genotyping of twelve KDR alleles in Ae. aegypti field populations, three PCR based assays were used; with DNA sequencing; ASPCR; PCR-RFLP. Genomic DNA was isolated, and three partial domains (II, III, and IV) of VGSC were amplified and sequenced. Results: Molecular screening for common KDR mutations, revealed the presence of five mutations viz. S989P, V1016G, T1520I, F1534C/L. Two novel mutations were observed, first at T1520 (ACC) residue where a C > T substitution at the second position of codon results in amino acid change to Isoleucine (ATC). Second mutation was an alternative point mutation at F1534 (TTC) residue where a substitution of T > C at the first position of codon results in an amino acid change to Leucine (CTC). ASPCRs were not accurate, so three PCR-RFLP assays were developed for genotyping of five KDR alleles in Ae. aegypti; viz. T1520I, F1534C/L. Representative samples of all genotypes (n=200) were sequenced to validate the newly developed PCR based assays for Ae. aegypti. Genotyping results showed that 989P is linked to 1016G and novel mutation 1520I was always found with 1534C allele. Conclusion: Present study confirmed the presence of DDT and pyrethroid resistance among Ae. aegypti populations in India and for the first time reported KDR mutations in this species from India including two novel mutations. Results of present study lead us to infer that, at least five KDR mutations (S989P, V1016G, T1530I, F1534C, and F1534L) can be seen as a potential marker for DDT/pyrethroid resistance.

Keywords: F1534C, F1534L, S989P, T1530I, V1016G

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913 Effect of Madecassoside on the Antioxidant Status of Streptozotocin-Nicotinamide Induced Diabetes in Sprague-Dawley Rats

Authors: C. Mayuren, C. K. Paul Wang, K. Purushotham, C. Dinesh Kumar

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Diabetes Mellitus (DM) is one of the most common non-communicable diseases globally. Although significant advances have led to better understanding of the condition and the development of effective therapies and preventive strategies, the pathway to cure remains elusive and DM prevails as a serious medical challenge in the 21st century. Oxidative stress has been suggested to contribute to the progression and pathophysiological conditions of diabetes. Madecassoside (MA) a major pentacyclic triterpenoid, has been demonstrated to possess various biological activities. However, no attempt has been made to study the antioxidant activity in diabetic rats. Therefore, the present study is aimed to evaluate the antioxidant effect of MA on streptozotocin-nicotinamide induced type-2 diabetes in Sprague-Dawley rats. The study protocol was approved by the institutional ethical committee prior to the conduct of research. Adult male Sprague-Dawley rats weighing 250-300 g were used in the study. The animals were rendered diabetic with a single intraperitoneal dose of streptozotocin (65 mg/kg) and nicotinamide (110 mg/kg). The diabetic animals after a stabilisation period of 14 days received various treatments (Madecassoside 50 mg/kg; Glimepiride 2.5 mg/kg) suspended in 0.5% carboxymethyl cellulose orally, for a period of 28 days. The animals fasted overnight after the last treatment were sacrificed and the pancreas, liver and kidneys were isolated. The weighted quantity of the samples of various treatments were homogenised in ice-cold condition and were subjected to lipid peroxidation, catalase and superoxide dismutase assay. The data’s obtained were subjected to statistical analysis. Diabetic rats showed significant increase in lipid peroxidation and decrease in enzymatic antioxidant levels. All the treated groups had significantly higher SOD, CAT and reduced LPO activity in the pancreas, liver and kidney. Results suggest madecassoside to have potential antioxidant effect against the diabetic model. However further investigations are necessary to study the mechanism at the cellular level.

Keywords: antioxidant, diabetes, madecassoside, nicotinamide, streptozotocin

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912 The Antibacterial and Anticancer Activity of Marine Actinomycete Strain HP411 Isolated in the Northern Coast of Vietnam

Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

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911 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

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Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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910 Microfluidic Chambers with Fluid Walls for Cell Biology

Authors: Cristian Soitu, Alexander Feuerborn, Cyril Deroy, Alfonso Castrejon-Pita, Peter R. Cook, Edmond J. Walsh

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Microfluidics now stands as an academically mature technology after a quarter of a century research activities have delivered a vast array of proof of concepts for many biological workflows. However, translation to industry remains poor, with only a handful of notable exceptions – e.g. digital PCR, DNA sequencing – mainly because of biocompatibility issues, limited range of readouts supported or complex operation required. This technology exploits the domination of interfacial forces over gravitational ones at the microscale, replacing solid walls with fluid ones as building blocks for cell micro-environments. By employing only materials used by biologists for decades, the system is shown to be biocompatible, and easy to manufacture and operate. The method consists in displacing a continuous fluid layer into a pattern of isolated chambers overlaid with an immiscible liquid to prevent evaporation. The resulting fluid arrangements can be arrays of micro-chambers with rectangular footprint, which use the maximum surface area available, or structures with irregular patterns. Pliant, self-healing fluid walls confine volumes as small as 1 nl. Such fluidic structures can be reconfigured during the assays, giving the platform an unprecedented level of flexibility. Common workflows in cell biology are demonstrated – e.g. cell growth and retrieval, cloning, cryopreservation, fixation and immunolabeling, CRISPR-Cas9 gene editing, and proof-of-concept drug tests. This fluid-shaping technology is shown to have potential for high-throughput cell- and organism-based assays. The ability to make and reconfigure on-demand microfluidic circuits on standard Petri dishes should find many applications in biology, and yield more relevant phenotypic and genotypic responses when compared to standard microfluidic assays.

Keywords: fluid walls, micro-chambers, reconfigurable, freestyle

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909 Growth and Nutrient Utilization of Some Citrus Peels and Vitamin Premix as Additives in Clarias Gariepinus Diets

Authors: Eunice Oluwayemisi Adeparusi, Mary Adedolapo Ijadeyila

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The study was carried out at the Federal University of Technology, Akure, Nigeria, West Africa. Seven set of diets were prepared comprising of two sets. The first set consisted of a combination of three diets from a combination of two different citrus peels from Orange (Citrus sinesis), Tangerine (Citrus tangerina / Citrus reticulata) and Tangelo (Citrus tangelo a hybrid of Citrus reticulata and Citrus maxima) at 50:50 while the other three consisted f50:50. Diet with 100% vitamin premix served as the control. Air-dried citrus peels were added in a 40% crude protein diet for the juveniles (4.49±0.05g) Clarias gariepinus. The experiment was carried out for a period of 56 days in triplicate trials. Fish were randomly distributed into twenty-one tanks at ten fish per tanks. The feed was extruded and fed to satiation twice daily. The result shows that fish fed Tangelo and Tangerine (TGL-TGR) had the best growth response in terms of final weight, specific growth rate, feed conversion ratio and feed utilization efficiency when compared with other diets. The FCR of fish in the diet ranges from 0.93-1.62. Fish fed the mixture of Orange peel and Vitamin-mineral premix (ORG-VIT) and those on Tangelo and Vitamin-mineral premix (TGL-VIT) had higher survival rate. There were significant differences (P<0.05) in the mean final weight, weight gain and specific growth rate. The result shows that citrus peels enhance the growth performance and feed utilization of the juvenile of African mud catfish, thereby reducing the cost of fish production.

Keywords: African mud catfish, growth, citrus peels, vitamin-mineral premix, nutrient utilization, additives

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908 Inactivation of Root-Knot Nematode Eggs Meloidogyne enterolobii in Irrigation Water Treated with Ozone

Authors: I. A. Landa-Fernandez, I. Monje-Ramirez, M. T. Orta-Ledesma

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Every year plant-parasitic nematodes diminish the yield of high-value crops worldwide causing important economic losses. Currently, Meloidogyne enterolobii has increased its importance due to its high aggressiveness, increasing geographical distribution and host range. Root-knot nematodes inhabit the rhizosphere soil around plant roots. However, they can come into contact with irrigation water. Thus, plant-parasitic nematodes can be transported by water, as eggs or juveniles. Due to their high resistance, common water disinfection methods are not effective for inactivating these parasites. Ozone is the most effective disinfectant for microbial inactivation. The objective of this study is to demonstrate that ozone treatment is an alternative method control in irrigation water of the root-knot nematode M. enterolobii. It has been shown that ozonation is an effective treatment for the inactivation of protozoan cysts and oocysts (Giardia and Cryptosporidium) and for other species of the genus Meloidogyne (M. incognita), but not for the enterolobii specie. In this study, the strain of M. enterolobii was isolated from tomatoes roots. For the tests, eggs were used and were inoculated in water with similar characteristics of irrigation water. Subsequently, the disinfection process was carried out in an ozonation unit. The performance of the treatments was evaluated through the egg's viability by assessing its structure by optical microscopy. As a result of exposure to ozone, the viability of the nematode eggs was reduced practically in its entirety; with dissolved ozone levels in water close to the standard concentration (equal to 0.4 mgO₃/L), but with high contact times (greater than 4 min): 0.2 mgO₃/L for 15 minutes or 0.55 mgO₃/L for 10 minutes. Additionally, the effect of temperature, alkalinity and organic matter of the water was evaluated. Ozonation is effective and a promising alternative for the inactivation of nematodes in irrigation water, which could contribute to diminish the agricultural losses caused by these organisms.

Keywords: inactivation process, irrigation water treatment, ozonation, plant-parasite nematodes

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907 Effects of Indole on Aerobic Biodegradation of Butanoic Acid by Pseudomonas aeruginosa and Serratia marcescens

Authors: J. B. J. Njalam’mano, E. M. N. Chirwa

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In low resource settings in Africa and other developing regions, pit latrines remain the dominant basic minimum acceptable form of sanitation. However, unpleasant smells-malodours emitted from faecal sludge in the pit latrines, which elicit disgusting or repulsive response, are one of the factors that thwart people to use latrines and instead opt for open defecation as an alternative. This provides an important but often overlooked major impediment, dissuading people from adopting and using the pit latrines hence affecting successful, effective sanitation promotion. The malodours are primarily attributed to four odorants: butanoic acid (C₄H₈O₂), dimethyl trisulphide (C₂H₆S₃), indole (C₈H₇N) and para-cresol (C₇H₈O). Several pit latrine deodorisation methods such as addition of carbonous materials, use of ventilation systems and urine separation are available, and they continue to occupy their niche, but social, economic, environmental and technological shortfalls remain. Bioremediation has been gaining popularity because it is inexpensive, simple to operate and environmentally friendly. Recently, the biodegradation of butanoic acid as individual odorant has been studied. However, to the best of our knowledge, there have been no kinetic studies of the butanoic acid in the presence of other key odorous compounds. In this study, a series of experiments were conducted to investigate the effects of indole on the removal of butanoic acid under aerobic conditions using indigenous bacteria strains, Pseudomonas aeruginosa, and Serratia marcescens isolated from faecal sludge as pure cultures as well as mixed cultures. In this purpose, butanoic acid removal was performed in a batch reactor containing the bacterial strains in mineral salt medium (MSM) amended with 3000 ppm of butanoic acid at the temperature of 30°C, under continuous stirring rate of 150 rpm and the concentration of indole was varied from 50-200 ppm. The initial pH of the solution was in the range of 6.0-7.2. Overall, there were significant differences in the bacterial growth rate and total butanoic acid removal dependent on the concentration of indole in the solution.

Keywords: biodegradation, butanoic acid, indole, pit latrine

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906 Development of Nanoparticulate Based Chimeric Drug Delivery System Using Drug Bioconjugated Plant Virus Capsid on Biocompatible Nanoparticles

Authors: Indu Barwal, Shloka Thakur, Subhash C. Yadav

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The plant virus capsid protein based nanoparticles are extensively studied for their application in biomedical research for development of nanomedicines and drug delivery systems. We have developed a chimeric drug delivery system by controlled in vitro assembly of separately bioconjugated fluorescent dye (as reporting molecule), folic acid (as receptor binding biomolecule for targeted delivery) and doxorubicin (as anticancer drug) using modified EDC NHS chemistry on heterologously overexpressed (E. coli) capsid proteins of cowpea chlorotic mottle virus (CCMV). This chimeric vehicle was further encapsidated on gold nanoparticles (20nm) coated with 5≠ thiolated DNA probe to neutralize the positive charge of capsid proteins. This facilitates the in vitro assembly of modified capsid subunits on the gold nanoparticles to develop chimeric GNPs encapsidated targeted drug delivery system. The bioconjugation of functionalities, number of functionality on capsid subunits as well as virus like nanoparticles, structural stability and in vitro assembly were confirmed by SDS PAGE, relative absorbance, MALDI TOF, ESI-MS, Circular dichroism, intrinsic tryptophan fluorescence, zeta particle size analyzer and TEM imaging. This vehicle was stable at pH 4.0 to 8.0 suitable for many organelles targeting. This in vitro assembled chimeric plant virus like particles could be suitable for ideal drug delivery vehicles for subcutaneous cancer treatment and could be further modified for other type of cancer treatment by conjugating other functionalities (targeting, drug) on capsids.

Keywords: chimeric drug delivery vehicles, bioconjugated plant, virus, capsid

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905 Quantification and Identification of the Main Components of the Biomass of the Microalgae Scenedesmus SP. – Prospection of Molecules of Commercial Interest

Authors: Carolina V. Viegas, Monique Gonçalves, Gisel Chenard Diaz, Yordanka Reyes Cruz, Donato Alexandre Gomes Aranda

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To develop the massive cultivation of microalgae, it is necessary to isolate and characterize the species, improving genetic tools in search of specific characteristics. Therefore, the detection, identification and quantification of the compounds that compose the Scenedesmus sp. were prerequisites to verify the potential of these microalgae. The main objective of this work was to carry out the characterization of Scenedesmus sp. as to the content of ash, carbohydrates, proteins and lipids as well as the determination of the composition of their lipid classes and main fatty acids. The biomass of Scenedesmus sp, showed 15,29 ± 0,23 % of ash and CaO (36,17 %) was the main component of this fraction, The total protein and carbohydrate content of the biomass was 40,74 ± 1,01 % and 23,37 ± 0,95 %, respectively, proving to be a potential source of proteins as well as carbohydrates for the production of ethanol via fermentation, The lipid contents extracted via Bligh & Dyer and in situ saponification were 8,18 ± 0,13 % and 4,11 ± 0,11 %, respectively. In the lipid extracts obtained via Bligh & Dyer, approximately 50 % of the composition of this fraction consists of fatty compounds, while the other half is composed of an unsaponifiable fraction composed mainly of chlorophylls, phytosterols and carotenes. From the lowest yield, it was possible to obtain a selectivity of 92,14 % for fatty components (fatty acids and fatty esters) confirmed through the infrared spectroscopy technique. The presence of polyunsaturated acids (~45 %) in the lipid extracts indicated the potential of this fraction as a source of nutraceuticals. The results indicate that the biomass of Scenedesmus sp, can become a promising potential source for obtaining polyunsaturated fatty acids, carotenoids and proteins as well as the simultaneous obtainment of different compounds of high commercial value.

Keywords: microalgae, Desmodesmus, lipid classes, fatty acid profile, proteins, carbohydrates

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904 Field Prognostic Factors on Discharge Prediction of Traumatic Brain Injuries

Authors: Mohammad Javad Behzadnia, Amir Bahador Boroumand

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Introduction: Limited facility situations require allocating the most available resources for most casualties. Accordingly, Traumatic Brain Injury (TBI) is the one that may need to transport the patient as soon as possible. In a mass casualty event, deciding when the facilities are restricted is hard. The Extended Glasgow Outcome Score (GOSE) has been introduced to assess the global outcome after brain injuries. Therefore, we aimed to evaluate the prognostic factors associated with GOSE. Materials and Methods: In a multicenter cross-sectional study conducted on 144 patients with TBI admitted to trauma emergency centers. All the patients with isolated TBI who were mentally and physically healthy before the trauma entered the study. The patient’s information was evaluated, including demographic characteristics, duration of hospital stays, mechanical ventilation on admission laboratory measurements, and on-admission vital signs. We recorded the patients’ TBI-related symptoms and brain computed tomography (CT) scan findings. Results: GOSE assessments showed an increasing trend by the comparison of on-discharge (7.47 ± 1.30), within a month (7.51 ± 1.30), and within three months (7.58 ± 1.21) evaluations (P < 0.001). On discharge, GOSE was positively correlated with Glasgow Coma Scale (GCS) (r = 0.729, P < 0.001) and motor GCS (r = 0.812, P < 0.001), and inversely with age (r = −0.261, P = 0.002), hospitalization period (r = −0.678, P < 0.001), pulse rate (r = −0.256, P = 0.002) and white blood cell (WBC). Among imaging signs and trauma-related symptoms in univariate analysis, intracranial hemorrhage (ICH), interventricular hemorrhage (IVH) (P = 0.006), subarachnoid hemorrhage (SAH) (P = 0.06; marginally at P < 0.1), subdural hemorrhage (SDH) (P = 0.032), and epidural hemorrhage (EDH) (P = 0.037) were significantly associated with GOSE at discharge in multivariable analysis. Conclusion: Our study showed some predictive factors that could help to decide which casualty should transport earlier to a trauma center. According to the current study findings, GCS, pulse rate, WBC, and among imaging signs and trauma-related symptoms, ICH, IVH, SAH, SDH, and EDH are significant independent predictors of GOSE at discharge in TBI patients.

Keywords: field, Glasgow outcome score, prediction, traumatic brain injury.

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903 The Retinoprotective Effects and Mechanisms of Fungal Ingredient 3,4-Dihydroxybenzalacetone through Inhibition of Retinal Müller and Microglial Activation

Authors: Yu-Wen Cheng, Jau-Der Ho, Liang-Huan Wu, Fan-Li Lin, Li-Huei Chen, Hung-Ming Chang, Yueh-Hsiung Kuo, George Hsiao

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Retina glial activation and neuroinflammation have been confirmed to cause devastating responses in retinodegenerative diseases. The expression and activation of matrix metalloproteinase (MMP)-9 and inducible nitric oxide synthase (iNOS) could be exerted as the crucial pathological factors in glaucoma- and blue light-induced retinal injuries. The present study aimed to investigate the retinoprotective effects and mechanisms of fungal ingredient 3,4-dihydroxybenzalacetone (DBL) isolated from Phellinus linteus in the retinal glial activation and retinodegenerative animal models. According to the cellular studies, DBL significantly and concentration-dependently abrogated MMP-9 activation and expression in TNFα-stimulated retinal Müller (rMC-1) cells. We found the inhibitory activities of DBL were strongly through the STAT- and ERK-dependent pathways. Furthermore, DBL dramatically attenuated MMP-9 activation in the stimulated Müller cells exposed to conditioned media from LPS-stimulated microglia BV-2 cells. On the other hand, DBL strongly suppressed LPS-induced production of NO and ROS and expression of iNOS in microglia BV-2 cells. Consistently, the phosphorylation of STAT was substantially blocked by DBL in LPS-stimulated microglia BV-2 cells. In the evaluation of retinoprotective functions, the high IOP-induced scotopic electroretinographic (ERG) deficit and blue light-induced abnormal pupillary light response (PLR) were assessed. The deficit scotopic ERG responses markedly recovered by DBL in a rat model of glaucoma-like ischemia/reperfusion (I/R)-injury. DBL also reduced the aqueous gelatinolytic activity and retinal MMP-9 expression in high IOP-injured conditions. Additionally, DBL could restore the abnormal PLR and reduce retinal MMP-9 activation. In summary, DBL could ameliorate retinal neuroinflammation and MMP-9 activation by predominantly inhibiting STAT3 activation in the retinal Müller cells and microglia, which exhibits therapeutic potential for glaucoma and other retinal degenerative diseases.

Keywords: glaucoma, blue light, DBL, retinal Müller cell, MMP-9, STAT, Microglia, iNOS, ERG, PLR

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902 Effect of Ecologic Fertilizers on Productivity and Yield Quality of Common and Spelt Wheat

Authors: Danutė Jablonskytė-Raščė, Audronė MankevičIenė, Laura Masilionytė

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During the period 2009–2015, in Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry, the effect of ecologic fertilizers Ekoplant, bio-activators Biokal 01 and Terra Sorb Foliar and their combinations on the formation of the productivity elements, grain yield and quality of winter wheat, spelt (Triticum spelta L.), and common wheat (Triticum aestivum L.) was analysed in ecological agro-system. The soil under FAO classification – Endocalcari-Endo-hypogleyic-Cambisol. In a clay loam soil, ecological fertilizer produced from sunflower hull ash and this fertilizer in combination with plant extracts and bio-humus exerted an influence on the grain yield of spelt and common wheat and their mixture (increased the grain yield by 10.0%, compared with the unfertilized crops). Spelt grain yield was by on average 16.9% lower than that of common wheat and by 11.7% lower than that of the mixture, but the role of spelt in organic production systems is important because with no mineral fertilization it produced grains with a higher (by 4%) gluten content and exhibited a greater ability to suppress weeds (by on average 61.9% lower weed weight) compared with the grain yield and weed suppressive ability of common wheat and mixture. Spelt cultivation in a mixture with common wheat significantly improved quality indicators of the mixture (its grain contained by 2.0% higher protein content and by 4.0% higher gluten content than common wheat grain), reduced disease incidence (by 2-8%), and weed infestation level (by 34-81%).

Keywords: common and spelt-wheat, ecological fertilizers, bio-activators, productivity elements, yield, quality

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901 Perspectives and Challenges a Functional Bread With Yeast Extract to Improve Human Diet

Authors: Jelena Filipović, Milenko Košutić, Vladimir Filipović

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In the last decades urban population is characterized by sedentary lifestyles, low physical activity and "fast food". These changes in diet and physical non activity have been associated with the increase of chronic non diseases. Bread is one of the most popularly wheat products consumed worldwide. Spelt wheat has shown potential in various food applications, including bread, pasta, breakfast cereal and other products of altered nutritional characteristics compared to conventional wheat products. It has very high protein content and even 30 to 60% higher concentration of mineral elements Fe, Zn, Cu, Mg and P compared to Triticum Aestivum. Spelt wheat is growing without the use of pesticides in harsh ecological conditions and it is an old cultivar. So it can be used for organic and health safe food. Changes in the formulation of bread with the aim to improve their nutritional and functional properties usually lead to changes in the dough properties which is related reflected to the quality of the finished product. The aim of this paper is researching the impact of adding yeast extract to bread on sensory characteristics and consumer acceptance of a new product as a key factor for successful marketing of a new product. The sensory analysis of bread with 5% yeast extract is as follows: the technological quality is very good (3.8) and the color of the product is excellent (4.85). Based on data consumers survey declared that they liked the taste of bread with 5% yeast extract (74%), consumers marked the product as likeable (70%), and 75% of the total number of respondents would buy this new product. This paper is promoting a new type of bread with 5% yeast extract (Z score 0.80) to improve diet and novel functional product which intended for consumers conscious about their health and diet.

Keywords: bread, yeast extract, sensory analysis, consumer survey, score analysis Z

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900 Association of Mir-196a Expression in Esophageal Tissue with Barrett´s Esophagus and Esophageal Adenocarcinoma

Authors: Petra Borilova Linhartova, Michaela Ruckova, Sabina Sevcikova, Natalie Mlcuchova, Jan Bohm, Katerina Zukalova, Monika Vlachova, Jiri Dolina, Lumir Kunovsky, Radek Kroupa, Zdenek Pavlovsky, Zdenek Danek, Tereza Deissova, Lydie Izakovicova Holla, Ondrej Slaby, Zdenek Kala

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Esophageal adenocarcinoma (EAC) is a highly aggressive malignancy that frequently develops from Barrett's esophagus (BE), a premalignant pathologic change occurring in the lower end of the esophagus. Specific microRNAs (miRNAs), small non-coding RNAs that function as posttranscriptional regulators of gene expression, were repeatedly proved to play key roles in the pathogenesis of these diseases. This pilot study aimed to analyze four selected miRNAs in esophageal tissues from healthy controls (HC) and patients with reflux esophagitis (RE)/BE/EAC, as well as to compare expression at the site of Barrett's mucosa/adenocarcinoma and healthy esophageal tissue outside the area of the main pathology in patients with BE/EAC. In this pilot study, 22 individuals (3 HC, 8 RE, 5 BE, 6 EAC) were included and endoscopically examined. RNA was isolated from the fresh-frozen esophageal tissue (stored in the RNAlater™ Stabilization Solution −70°C) using the AllPrep DNA/RNA/miRNA Universal Kit. Subsequent RT-qPCR analysis was performed using selected TaqMan MicroRNA Assays for miR-21, miR-34a, miR-196a, miR-196b, and endogenous control (RNU44). While the expression of miR-21 in the esophageal tissue with the main pathology was decreased in BE and EAC patients in comparison to the group of HC and RE patients (p=0.01), the expression of miR-196a was increased in the BE and EAC patients (p<0.01). Correlations between those miRNAs expression in tissue and severity of diagnosis were observed (p<0.05). In addition, miR-196a was significantly more expressed at the site with the main pathology than in paired adjacent esophageal tissue in BE and EAC patients (p<0.01). In conclusion, our pilot results showed that miR-196a, which regulates the proliferation, invasion, and migration (and was previously associated with esophageal squamous cell carcinoma and marked as a potential therapeutic target), could be a diagnostic tissue biomarker for BE and EAC as well.

Keywords: microRNA, barrett´s esophagus, esophageal adenocarcinoma, biomarker

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899 Identification of Bioactive Metabolites from Ficus carica and Their Neuroprotective Effects of Alzheimer's Disease

Authors: Hanan Khojah, RuAngelie Edrada-Ebel

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Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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898 Malignant Idiopathic Intracranial Hypertension Revealed a Hidden Primary Spinal Leptomeningeal Medulloblastoma

Authors: Naim Izet Kajtazi

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Context: Frequently, the cause of raised intracranial pressure remains unresolved and rarely is related to spinal tumors, moreover less to spinal medulloblastoma without primary brain focus. Process: An 18-year-old woman had a 3-month history of headaches and impaired vision. Neurological examination revealed bilateral sixth cranial nerve palsies with bilateral papilloedema of grade III. No focal brain or spine lesion was found on imaging. Consecutive lumbar punctures showed high opening pressure and subsequent increasing protein level. The meningeal biopsy was negative. At one point, she developed an increasing headache, vomiting and back pain. Spine MRI showed diffuse nodular leptomeningeal enhancement with the largest nodule at T6–T7. Malignant cells were detected in cerebrospinal fluid. She underwent laminectomy with excisional biopsy, and pathology showed medulloblastoma WHO grade IV. Outcome: She was treated with chemotherapy and craniospinal irradiation and made a good recovery. Relevance: Primary spinal leptomeningeal medulloblastoma is extremely rare, especially without primary brain focus, but may cause increased intracranial pressure, even in the early microscopic phases, and it should be considered in the differential diagnosis if conventional and aggressive treatment of idiopathic intracranial hypertension fails. We assume that arachnoiditis from tumor seeding caused increased intracranial pressure. Appropriate neurosurgical intervention and surgical biopsy are mandated if a suspicious lesion is detected. Consider proper rescreening of the whole neuroaxis in refractory cases of intracranial hypertension.

Keywords: CNS infection, IIH, headache, primary spinal leptomeningeal medulloblastoma

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897 Analysis of the Impact of Suez Canal on the Robustness of Global Shipping Networks

Authors: Zimu Li, Zheng Wan

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The Suez Canal plays an important role in global shipping networks and is one of the most frequently used waterways in the world. The 2021 canal obstruction by ship Ever Given in March 2021, however, completed blocked the Suez Canal for a week and caused significant disruption to world trade. Therefore, it is very important to quantitatively analyze the impact of the accident on the robustness of the global shipping network. However, the current research on maritime transportation networks is usually limited to local or small-scale networks in a certain region. Based on the complex network theory, this study establishes a global shipping complex network covering 2713 nodes and 137830 edges by using the real trajectory data of the global marine transport ship automatic identification system in 2018. At the same time, two attack modes, deliberate (Suez Canal Blocking) and random, are defined to calculate the changes in network node degree, eccentricity, clustering coefficient, network density, network isolated nodes, betweenness centrality, and closeness centrality under the two attack modes, and quantitatively analyze the actual impact of Suez Canal Blocking on the robustness of global shipping network. The results of the network robustness analysis show that Suez Canal blocking was more destructive to the shipping network than random attacks of the same scale. The network connectivity and accessibility decreased significantly, and the decline decreased with the distance between the port and the canal, showing the phenomenon of distance attenuation. This study further analyzes the impact of the blocking of the Suez Canal on Chinese ports and finds that the blocking of the Suez Canal significantly interferes withChina's shipping network and seriously affects China's normal trade activities. Finally, the impact of the global supply chain is analyzed, and it is found that blocking the canal will seriously damage the normal operation of the global supply chain.

Keywords: global shipping networks, ship AIS trajectory data, main channel, complex network, eigenvalue change

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896 Investigate the Side Effects of Patients With Severe COVID-19 and Choose the Appropriate Medication Regimens to Deal With Them

Authors: Rasha Ahmadi

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In December 2019, a coronavirus, currently identified as SARS-CoV-2, produced a series of acute atypical respiratory illnesses in Wuhan, Hubei Province, China. The sickness induced by this virus was named COVID-19. The virus is transmittable between humans and has caused pandemics worldwide. The number of death tolls continues to climb and a huge number of countries have been obliged to perform social isolation and lockdown. Lack of focused therapy continues to be a problem. Epidemiological research showed that senior patients were more susceptible to severe diseases, whereas children tend to have milder symptoms. In this study, we focus on other possible side effects of COVID-19 and more detailed treatment strategies. Using bioinformatics analysis, we first isolated the gene expression profile of patients with severe COVID-19 from the GEO database. Patients' blood samples were used in the GSE183071 dataset. We then categorized the genes with high and low expression. In the next step, we uploaded the genes separately to the Enrichr database and evaluated our data for signs and symptoms as well as related medication regimens. The results showed that 138 genes with high expression and 108 genes with low expression were observed differentially in the severe COVID-19 VS control group. Symptoms and diseases such as embolism and thrombosis of the abdominal aorta, ankylosing spondylitis, suicidal ideation or attempt, regional enteritis were observed in genes with high expression and in genes with low expression of acute and subacute forms of ischemic heart, CNS infection and poliomyelitis, synovitis and tenosynovitis. Following the detection of diseases and possible signs and symptoms, Carmustine, Bithionol, Leflunomide were evaluated more significantly for high-expression genes and Chlorambucil, Ifosfamide, Hydroxyurea, Bisphenol for low-expression genes. In general, examining the different and invisible aspects of COVID-19 and identifying possible treatments can help us significantly in the emergency and hospitalization of patients.

Keywords: phenotypes, drug regimens, gene expression profiles, bioinformatics analysis, severe COVID-19

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895 Prevalence, Antimicrobial Susceptibility Pattern and Associated Risk Factors for Salmonella Species and Escherichia coli from Raw Meat at Butchery Houses in Mekelle, Tigray, Ethiopia

Authors: Haftay Abraha Tadesse, Atsebaha Gebrekidan Kahsay, Mahumd Abdulkader

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Background: Salmonella species and Escherichia coli are important foodborne pathogens affecting humans and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. This study was aimed to determine the prevalence, antimicrobial susceptibility patterns and associated risk factors for Salmonella species and E. coli in raw meat from butchery houses of Mekelle, Northern Ethiopia. Methodology: A cross-sectional study was conducted from January to September 2019. Socio-demographic data and risk factors were collected using a predesigned questionnaire. Meat samples were collected aseptically from the butchery houses and transported using icebox to Mekelle University, College of Veterinary Sciences for the isolation and identification of Salmonella species and E. coli, Antimicrobial susceptibility patterns were determined using Kirby disc diffusion method. Data obtained were cleaned and entered into Statistical Package for the Social Sciences version 22 and logistic regression models with odds ratio were calculated. P-value < 0.05 was considered as statistically significant. Results: A total of 153 out of 384 (39.8%) of the meat specimens were found to be contaminated. The contamination of Salmonella species and E. coli were 15.6% (n=60) and 20.8%) (n=80), respectively. Mixed contamination (Salmonella species and E. coli) was observed in 13 (3.4 %) of the analyzed. Poor washing hands regularly (AOR = 8.37; 95% CI: 2.75-25.50) and not using gloves during meat handling (AOR=11. 28; 95% CI: (4.69 27.10) were associated with an overall bacterial contamination.About 95.5% of the tested isolates were sensitive to chloramphenicol and norfloxacin while the resistance of amoxyclav_amoxicillin and erythromycin were both isolated bacteria species. The overall multidrug resistance pattern for Salmonella and E. coli were 51.4% (n=19) and 31.8% (14), respectively. Conclusion: Of the 153 (153/384) contaminated raw meat, 60 (15.6%) and 80 (20.8%) were contaminated by Salmonella species and E. coli, respectively. Poor hand washing practice and not using glove during meat handling showed significant association with bacterial contamination. Multidrug-resistant showed in Salmonella species and E. coli were 19 (51.4%) and 14 (31.8%), respectively.

Keywords: antimicrobial susceptibility test, butchery houses, e. coli, salmonella species

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894 Calpain-Mediated, Cisplain-Induced Apoptosis in Breast Cancer Cells

Authors: Shadia Al-Bahlani, Khadija Al-Bulushi, Zuweina Al-Hadidi, Buthaina Al-Dhahl, Nadia Al-Abri

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Breast cancer is the most common cancer in women worldwide. Triple-Negative Breast Cancer (TNBC) is an aggressive type of breast cancer, which is defined by the absence of Estrogen (ER), Progesterone (PR) and human epidermal growth factor (Her-2) receptors. The calpain system plays an important role in many cellular processes including apoptosis, necrosis, cell signaling and proliferation. However, the role of calpain in cisplatin (CDDP)-induced apoptosis in TNBC cells is not fully understood. Here, TNBC (MDA-MB231) cells were treated with different concentration of CDDP (0, 20 & 40 µM) and calpain activation and apoptosis were measured by western blot and Hoechst Stain respectively. In addition, calpain modulation by either activation and/or inhibition and its effect on CDDP-induced apoptosis were assessed by the same above approaches. Our findings showed that CDDP induced endoplasmic reticulum stress and thus Calcium release and subsequently activate calpain α-fodrin cleavage indicated by the increase in GRP78 and Calmodulin protein expression and respectively in MDA-MB231 cells. It also induced apoptosis as measured by Hoechst stain and caspase-12 cleavage. Calpain activation by both Cyclopiazonic acid and Thapsigargin showed similar effect and enhanced the sensitivity of these cells to CDDP treatment. On the other hand, calpain inhibition by either specific siRNA and/or exogenous inhibitor (Calpeptin) had an adverse effect where it attenuated calpain activation and thus CDDP- induced apoptosis in these cells. Altogether, these findings suggested that calpain activation play an essential role in sensitizing the TNBC cells to CDDP-induced apoptosis. This might lead to the discovery of novel treatment to over this aggressive type of breast cancer.

Keywords: calpain, cisplatin, apoptosis, breast cancer

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893 Early and Mid-Term Results of Anesthetic Management of Minimal Invasive Coronary Artery Bypass Grafting Using One Lung Ventilation

Authors: Devendra Gupta, S. P. Ambesh, P. K Singh

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Introduction: Minimally invasive coronary artery bypass grafting (MICABG) is a less invasive method of performing surgical revascularization. Minimally invasive direct coronary artery bypass (MIDCAB) provides many anesthetic challenges including one lung ventilation (OLV), managing myocardial ischemia, and pain. We present an early and midterm result of the use of this technique with OLV. Method: We enrolled 62 patients for analysis operated between 2008 and 2012. Patients were anesthetized and left endobronchial tube was placed. During the procedure left lung was isolated and one lung ventilation was maintained through right lung. Operation was performed utilizing off pump technique of coronary artery bypass grafting through a minimal invasive incision. Left internal mammary artery graft was done for single vessel disease and radial artery was utilized for other grafts if required. Postoperative ventilation was done with single lumen endotracheal tube. Median follow-up is 2.5 years (6 months to 4 years). Results: Median age was 58.5 years (41-77) and all were male. Single vessel disease was present in 36, double vessel in 24 and triple vessel disease in 2 patients. All the patients had normal left ventricular size and function. In 2 cases difficulty were encounter in placement of endobronchial tube. In 1 case cuff of endobronchial tube was ruptured during intubation. High airway pressure was developed on OLV in 1 case and surgery was accomplished with two lung anesthesia with low tidal volume. Mean postoperative ventilation time was 14.4 hour (11-22). There was no perioperative and 30 day mortality. Conversion to median sternotomy to complete the operation was done in 3.23% (2 out of 62 patients). One patient had acute myocardial infarction postoperatively and there were no deaths during follow-up. Conclusion: MICABG is a safe and effective method of revascularization with OLV in low risk candidates for coronary artery bypass grafting.

Keywords: MIDCABG, one lung ventilation, coronary artery bypass grafting, endobronchial tube

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