Search results for: recombinant protein purification

Authors: Radovan Čobanović, Milica Rankov Šicar

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New attitudes about nutrition impose new styles, and therefore a neNew attitudes about nutrition impose new styles, and therefore a new kind of food. The goal of our work was to define the shelf life of new extruded soya product with minimum 65% of protein based on the analyses. According to the plan it was defined that a certain quantity of the same batch of new product (soybean flakes) which had predicted shelf life of 2 years had to be stored for 24 months in storage and analyzed at the beginning and end of sustainability plan on instrumental analyses (heavy metals, pesticides and mycotoxins) and every month on sensory analyses (odor, taste, color, consistency), microbiological analyses (Salmonella spp., Escherichia coli, Enterobacteriaceae, sulfite-reducing clostridia, Listeria monocytogenes), chemical analyses (protein, ash, fat, crude cellulose, granulation) and at the beginning on GMO analyses. All analyses were tested according to: sensory analyses ISO 6658, Salmonella spp ISO 6579, Escherichia coli ISO 16649-2, Enterobacteriaceae ISO 21528-2, sulfite-reducing clostridia ISO 15213 and Listeria monocytogenes ISO 11290-2, chemical and instrumental analyses Serbian ordinance on the methods of physico-chemical analyses and GMO analyses JRC Compendium. The results obtained after the analyses which were done according to the plan during the 24 months indicate that are no changes of products concerning both sensory and chemical analyses. As far as microbiological results are concerned Salmonella spp was not detected and all other quantitative analyses showed values <10 cfu/g. The other parameters for food safety (heavy metals, pesticides and mycotoxins) were not present in analyzed samples and also all analyzed samples were negative concerning genetic testing. On the basis of monitoring the sample under defined storage conditions and analyses of quality control, GMO analyses and food safety of the sample during the shelf within two years, the results showed that all the parameters of the sample during defined period is in accordance with Serbian regulative so that indicate that predicted shelf life can be adopted.w kind of food. The goal of our work was to define the shelf life of new extruded soya product with minimum 65% of protein based on the analyses. According to the plan it was defined that a certain quantity of the same batch of new product (soybean flakes) which had predicted shelf life of 2 years had to be stored for 24 months in storage and analyzed at the beginning and end of sustainability plan on instrumental analyses (heavy metals, pesticides and mycotoxins) and every month on sensory analyses (odor, taste, color, consistency), microbiological analyses (Salmonella spp., Escherichia coli, Enterobacteriaceae, sulfite-reducin clostridia, Listeria monocytogenes), chemical analyses (protein, ash, fat, crude cellulose, granulation) and at the beginning on GMO analyses. All analyses were tested according: sensory analyses ISO 6658, Salmonella spp ISO 6579, Escherichia coli ISO 16649-2, Enterobacteriaceae ISO 21528-2, sulfite-reducing clostridia ISO 15213 and Listeria monocytogenes ISO 11290-2, chemical and instrumental analyses Serbian ordinance on the methods of physico-chemical analyses and GMO analyses JRC Compendium. The results obtained after the analyses which were done according to the plan during the 24 months indicate that are no changes of products concerning both sensory and chemical analyses. As far as microbiological results are concerned Salmonella spp was not detected and all other quantitative analyses showed values <10 cfu/g. The other parameters for food safety (heavy metals, pesticides and mycotoxins) were not present in analyzed samples and also all analyzed samples were negative concerning genetic testing. On the basis of monitoring the sample under defined storage conditions and analyses of quality control, GMO analyses and food safety of the sample during the shelf within two years, the results showed that all the parameters of the sample during defined period is in accordance with Serbian regulative so that indicate that predicted shelf life can be adopted.

Keywords: extruded soya product, food safety analyses, GMO analyses, shelf life

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Authors: Govindan Sudha, Mathumitha Subramaniam, Alamelu Govindasamy, Sasikala Gunasekaran

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The aim of this study was to investigate the protective effect of Hypsizygus ulmarius polysaccharides (HUP) on reducing oxidative stress, cognitive impairment and neurotoxicity in D-galactose induced aging mice. Mice were subcutaneously injected with D-galactose (150 mg/kg per day) for 6 weeks and were administered HUP simultaneously. Aged mice receiving vitamin E (100 mg/kg) served as positive control. Chronic administration of D-galactose significantly impaired cognitive performance oxidative defence and mitochondrial enzymes activities as compared to control group. The results showed that HUP (200 and 400 mg/kg) treatment significantly improved the learning and memory ability in Morris water maze test. Biochemical examination revealed that HUP significantly increased the decreased activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), mitochondrial enzymes-NADH dehydrogenase, malate dehydrogenase (MDH), isocitrate dehydrogenase (ICDH), Na+K+, Ca2+, Mg2+ATPase activities, elevated the lowered total anti-oxidation capability (TAOC), glutathione (GSH), vitamin C and decreased the raised acetylcholinesterase (AChE) activities, malondialdehyde (MDA), hydroperoxide (HPO), protein carbonyls (PCO), advanced oxidation protein products (AOPP) levels in brain of aging mice induced by D-gal in a dose-dependent manner. In conclusion, present study highlights the potential role of HUP against D-galactose induced cognitive impairment, biochemical and mitochondrial dysfunction in mice. In vitro studies on the effect of HUP on scavenging DPPH, ABTS, DMPD, OH radicals, reducing power, B-carotene bleaching and lipid peroxidation inhibition confirmed the free radical scavenging and antioxidant activity of HUP. The results suggest that HUP possesses anti-aging efficacy and may have potential in treatment of neurodegenerative diseases.

Keywords: aging, antioxidants, mushroom, neurotoxicity

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Authors: Manjusha Choudhary

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Objective: Cassia occidentalis Linn. belongs to Family Caesalpiniaceae is a common weed scattered from the foothills of Himalayas to West Bengal, South India, Burma, and Sri Lanka. It is used widely in folklore medicine in India as laxative, expectorant, analgesic, anti-malarial, hepatoprotective, relaxant, anti-inflammatory and antidiabetic. The present study was carried out to investigate the hypoglycaemic and hypolipidemic activities of ethanolic extract of Cassia occidentalis stem bark. Methods: Stem bark extract of Cassia occidentalis (SBCO) was administered orally at 250 and 500 mg/kg doses to normal and streptozotocin (STZ) induced type-2 diabetic mice. Various parameters like fasting blood glucose (FBG) level, serum cholesterol, high density lipoprotein (HDL) cholesterol, triglycerides (TG), total protein, urea, creatinine, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) levels and physical parameters like change in body weight, food intake, water intake were performed for the evaluation of antidiabetic effects. Results: Both the doses of extract caused a marked decrease in FBG levels in STZ induced type 2 diabetic mice. Administration of SBCO led to the decrease in the blood glucose, food intake, water intake, organ weight, SGOT, SGPT levels with significant value and increased the levels of TG, HDL cholesterol, creatinine, cholesterol, total protein with a significant value (p < 0.05-0.01). The decrease in body weight induced by STZ was restored to normal with a significant value (p < 0.01) at both doses. Conclusion: Present study reveals that SBCO possess potent hypoglycaemic and hypolipidemic activities and supports the folklore use of the stem bark of plant as antidiabetic agent.

Keywords: Cassia occidentalis, diabetes, folklore, herbs, hypoglycemia, streptozotocin

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Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández

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Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.

Keywords: immunology, vaccines, pathogens, infectious disease

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Authors: Nuray Ucar, Pelin Altay, Ilkay Ozsev Yuksek

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Graphene oxide fibers have recently received increasing attention due to their excellent properties such as high specific surface area, high mechanical strength, good thermal properties and high electrical conductivity. They have shown notable potential in various applications including batteries, sensors, filtration and separation and wearable electronics. Carbon nanotubes (CNTs) have unique structural, mechanical, and electrical properties and can be used together with graphene oxide fibers for several application areas such as lithium ion batteries, wearable electronics, etc. Metals salts that can be converted into metal ions and metal oxide can be also used for several application areas such as battery, purification natural gas, filtration, absorption. This study investigates the effects of CNT and metal complex compounds (MnCl₂, metal salts) on the morphological structure of graphene oxide fibers. The graphene oxide dispersion was manufactured by modified Hummers method, and continuous graphene oxide fibers were produced with wet spinning. The CNT and MnCl₂ were incorporated into the coagulation baths during wet spinning process. Produced composite continuous fibers were analyzed with SEM, SEM-EDS and AFM microscopies and as spun fiber counts were measured.

Keywords: continuous graphene oxide fiber, Hummers' method, CNT, MnCl₂

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Authors: Mulugeta Gurum Gerechal

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Nowadays, the photocatalytic mechanism of water purification using nanoparticles has gained wider acceptance. For this purpose, the crystal form of N- TiO₂ and Ag-TiO₂ was prepared from TiCl₄, urea, NH₄OH, and AgNO₃ by sol-gel method and simple solid phase reaction followed by calcination at a temperature of 400°C for 4h at each. The synthesized photocatalysts were characterized using XRD, SEM, and UV-visible diffuse reflectance spectra. In the experiment, it was found that the absorption edge of N-TiO₂ was an efficient shift to visible light as compared to Ag-TiO₂. The XRD diffraction makes the particle size of N-TiO₂ smaller than Ag-TiO₂. The effect of catalyst loading and the effect of temperature on the photocatalytic efficiency of the prepared samples was tested using methylene blue as a target pollutant. The photocatalytic degradation efficiency of the catalysts for methylene blue was increased from 57.05 to 96.02% under solar radiation as the amount of the catalyst increased from 0.15 to 0.45 gram for N-TiO₂. Similarly, photocatalytic degradation of methylene blue was increased from 40.32 to 81.21% as the amount of Ag-TiO₂ increased from 0.05g to 0.1g. In addition, the photocatalytic degradation efficiency of the catalysts for the removal of methylene blue was increased from 58.00 to 98.00 and 47.00 to 81.21% under solar radiation as the calcination temperature of the catalyst increased from 300 to 500 for N-TiO₂ for Ag-TiO₂ 300 to 400⁰C. However, a further increase in catalyst loading and calcination temperature was found to decrease the degradation efficiency.

Keywords: photocatalysis, degradation, nanoparticles, catalyst loading, calcination, methylene blue

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Authors: Nadeem A. Siddique, Mohd Mujeeb, Kahkashan

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Introduction: The objective of the projected work is to study the occurrence of the aflatoxins B1, B2, G1and G2 in remedial plants, exclusively in Delphinium denudatum. The aflatoxins were analysed by high-performance liquid chromatography–tandem quadrupole mass spectrometry with electrospray ionization (HPLC–MS/MS) and immunoaffinity column chromatography were used for extraction and purification of aflatoxins. PDA media was selected for fungal count. Results: A good quality linear relationship was originated for AFB1, AFB2, AFG1 and AFG2 at 1–10 ppb (r > 0.9995). The analyte precision at three different spiking levels was 88.7–109.1 %, by means of low per cent relative standard deviations in each case. Within 5 to7 min aflatoxins can be separated using an Agilent XDB C18-column. We found that AFB1 and AFB2 were not found in D. denudatum. This was reliable through exceptionally low figures of fungal colonies observed after 6 hr of incubation. The developed analytical method is straightforward, be successfully used to determine the aflatoxins. Conclusion: The developed analytical method is straightforward, simple, accurate, economical and can be successfully used to find out the aflatoxins in remedial plants and consequently to have power over the quality of products. The presence of aflatoxin in the plant extracts was interrelated to the least fungal load in the remedial plants examined.

Keywords: aflatoxins, delphinium denudatum, liquid chromatography, mass spectrometry

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Authors: Ravi Kumar

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Agricultural wastes are the non-product outcomes of agricultural processing whose monetary value is less as compared to its collection cost, transportation, and processing. When such agricultural waste is not properly disposed of, it may damage the natural environment and cause detrimental pollution in the atmosphere. Agricultural development and intensive farming methods usually result in wastes that remarkably affect the rural environments in particular and the global environment in general. Agricultural waste has toxicity latent to human beings, animals, and plants through various indirect and direct outlets. The present paper explores the various activities that result in agricultural waste and the routes that can utilize the agricultural waste in a manageable manner to reduce its adverse impact on the environment. Presently, the agricultural waste management system for ecological agriculture and sustainable development has emerged as a crucial issue for policymakers. There is an urgent need to consider agricultural wastes as prospective resources rather than undesirable in order to avoid the transmission and contamination of water, land, and air resources. Waste management includes the disposal and treatment of waste with a view to eliminate threats of waste by modifying the waste to condense the microbial load. The study concludes that proper waste utilization and management will facilitate the purification and development of the ecosystem and provide feasible biofuel resources. This proper utilization and management of these wastes for agricultural production may reduce their accumulation and further reduce environmental pollution by improving environmental health.

Keywords: agricultural waste, utilization, management, environment, health

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Authors: Jinhee Kang, Kwangman Park, Ruhee Song, Suhee Kim, Do-Hyeon Yu, Kyoungseong Choi, Jinho Park

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Statement of the Problem: Animal blood components maintain homeostasis when animals are healthy, and changes in chemical composition of the blood and body fluids can be observed if animals have a disease. In particular, newborn calves are susceptible to disease and therefore hematologic tests and serum chemistry tests could become an important guideline to the diagnosis and the treatment of diseases. Diarrhea in newborn calves is the most damaging to cattle ranch, whether dairy or cattle fattening, and is a large part of calf atrophy and death. However, since the study on calf electrophoresis was not carried out, a survey analysis was conducted on it. Methodology and Theoretical Orientation: The calves were divided into healthy calves and disease (diarrhea) calves, and calves were classified by 1-14d, 15-28d, and more than 28d, respectively. The fecal state was classified by solid (0-value), semi-solid (1-value), loose (2-value) and watery (3-value). In the solid (0-value) and semi-solid (1-value) feces valuable pathogen was not detected, but loose (2-value) and watery (3-value) feces were detected. Findings: ALB, α-1, α-2, α-SUM, β and γ (Gamma) were examined by electrophoresis analysis of healthy calves and diarrhea calves. Test results showed that there were age differences between healthy calves and diarrheic calves. When we look at the γ-globulin at 1-14 days of age, we can see that the average calf of healthy calves is 16.8% and the average of diarrheal calves is 7.7%, when we look at the figures for the α-2 at 1-14 days, we found that healthy calves average 5.2% and diarrheal calves 8.7% higher than healthy cows. On α-1, 15-28 days, and after 28 days, healthy calves average 10.4% and diarrheal calves average 7.5% diarrhea calves were 12.6% and 12.4% higher than healthy calves. In the α-SUM, the healthy calves were 21.6%, 16.8%, and 14.5%, respectively, after 1-14 days, 15-28 days and 28 days. diarrheal calves were 23.1%, 19.5%, and 19.8%. Conclusion and Significance: In this study, we examined the electrophoresis results of healthy calves and diseased (diarrhea) calves, gamma globulin at 1-14 days of age were lower than those of healthy calves (diarrhea), indicating that the calf was unable to consume colostrum from the mother when it was a new calf. α-1, α-2, α-SUM may be associated with an acute inflammatory response as a result of increased levels of calves with diarrhea (diarrhea). Further research is needed to investigate the effects of acute inflammatory responses on additional calf-forming proteins. Information on the results of the electrophoresis test will be provided where necessary according to the item.

Keywords: alpha, electrophoretogram, serum protein, γ, gamma

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Authors: S. El Hasini, M. El Azzouzi, M. De Nobili, K. Azim, A. Zouahri

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Soil salinization is one of the most severe environmental hazards which threaten sustainable agriculture in arid and semi-arid regions, including Morocco. In this regard the application of organic matter to saline soil has confirmed its effectiveness. The present study was aimed to examine the effect of humic acid which represent, among others, the important component of organic matter that contributes to reduce soil salinity. In fact, different composts taken from Agadir (Morocco), with different C/N ratio, were tested. After extraction and purification of humic acid, the interaction with Na2CO3 was carried out. The reduction of salinity is calculated as a value expressed in mg Na2CO3 equivalent/g HA. The results showed that humic acid had generally a significant effect on salinity. In that respect, the hypothesis proposed that carboxylic groups of humic acid create bonds with excess sodium in the soil to form a coherent complex which descends by leaching operation. The comparison between composts was based on C/N ratio, it showed that the compost with the lower ratio C/N had the most important effect on salinity reduction, whereas the compost with higher C/N ratio was less effective. The study is attended also to evaluate the quality of each compost by determining the humification index, we noticed that the compost which have the lowest C/N (20) ratio was relatively less stable, where a greater predominance of the humified substances, when the compost with C/N ratio is 35 exhibited higher stability.

Keywords: compost, humic acid, organic matter, salinity

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Authors: P. Annapurna, Cecil Ross, Sudhir Krishna, Sweta Srivastava

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Irradiation plays a pivotal role in cervical cancer treatment, however some tumors exhibit resistance to therapy while some exhibit relapse, due to better repair and enhanced resistance mechanisms operational in their cells. The present study aims to understand the signaling mechanism operational in resistance phenotype and in the present study we report the role of Rho GTPase associated protein kinase (ROCK) signaling in cervical carcinoma radio-resistance. ROCK signaling has been implicated in several tumor progressions and is important for DNA repair. Irradiation of spheroid cultures of SiHa cervical carcinoma derived cell line at 6Gy resulted in generation of resistant cells in vitro which had better clonogenic abilities and formed larger and more colonies, in soft agar colony formation assay, as compared to the non-irradiated cells. These cells also exhibited an enhanced motility phenotype. Cell cycle profiling showed the cells to be blocked in G2M phase with enhanced pCDC2 levels indicating onset of possible DNA repair mechanism. Notably, 3 days post-irradiation, irradiated cells showed increased ROCK2 translocation to the nucleus with enhanced protein expression as compared to the non-irradiated cells. Radio-sensitization of the resistant cells was enhanced using Y27632, an inhibitor to ROCK signaling. The treatment of resistant cells with Y27632 resulted in increased cell death upon further irradiation. This observation has been confirmed using inhibitory antibodies to ROCK1/2. Result show that both ROCK1/2 have a functional contribution in radiation resistance of cervical cancer cells derived from cell lines. Interestingly enrichment of stem like cells (Hoechst negative cells) was also observed upon irradiation and these cells were markedly sensitive to Y27632 treatment. Our results thus suggest the role of ROCK signaling in radio-resistance in cervical carcinoma. Further studies with human biopsies, mice models and mechanistic of ROCK signaling in the context of radio-resistance will clarify the role of this molecule further and allow for therapeutics development.

Keywords: cervical carcinoma, radio-resistance, ROCK signaling, cancer treatment

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Authors: Fei Sun, Meilan Xu, Jianghui Zhu, Maria Stefanie Dwiyanti, Cheolwoo Park, Fanjiang Kong, Baohui Liu, Tetsuya Yamada, Jun Abe

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Reduced or lack of sensitivity to long daylengths is a key character for soybean, a short-day crop, to adapt to higher latitudinal environments. However, the photoperiod-insensitivity often results in a reduction of the duration of vegetative growth and final yield. To overcome this limitation, a photoperiod insensitive line (RIL16) was developed in this study that delayed flowering from the recombinant inbred population derived from a cross between a photoperiod-insensitive cultivar AGS292 and a late-flowering Thai cultivar K3. Expression analyses under SD and LD conditions revealed that the expression levels of FLOWERING LOCUS T (FT) orthologues, FT2a and FT5a, were lowered in RIL16 relative to AGS292, although the expression of E1, a soybean-specific suppressor for FTs, was inhibited in both conditions. A soybean orthologue of TARGET OF EAT1 (TOE1), another suppressor of FT, showed an upregulated expression in RIL16, which appeared to reflect a lower expression of miR172a. Our data suggest that the delayed flowering of RIL16 most likely is controlled by genes involved in an age-dependent pathway in flowering. The QTL analysis based on 1,125 SNPs obtained from Restriction Site Associated DNA Sequencing revealed two major QTLs for flowering dates in Chromosome 16 and two minor QTLs in Chromosome 4, all of which accounted for 55% and 48% of the whole variations observed in natural day length and artificially-induced long day length conditions, respectively. The intervals of the major QTLs harbored FT2a and FT5a, respectively, on the basis of annotated genes in the Williams 82 reference genome. Sequencing analysis further revealed a nonsynonymous mutation in FT2a and an SNP in the 3′ UTR region of FT5a. A further study may elucidate a detailed mechanism underlying the QTL for late flowering. The alleles from K3 at the two QTLs can be used singly or in combination to retain an appropriate duration of vegetative growth to maximize the final yield of photoperiod-insensitive soybeans.

Keywords: FT genes, miR72a, photoperiod-insensitive, soybean flowering

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Authors: German Hernandez-Alonso, Antonio Lazcano, Arturo Becerra

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Until today, viruses remain controversial and poorly understood; about their origin, this problem represents an enigma and one of the great challenges for the contemporary biology. Three main theories have tried to explain the origin of viruses: regressive evolution, escaped host gene, and pre-cellular origin. Under the perspective of the escaped host gene theory, it can be assumed a cellular origin of viral components, like protein RNA-binding domains. These universal distributed RNA-binding domains are related to the RNA metabolism processes, including transcription, processing, and modification of transcripts, translation, RNA degradation and its regulation. In the case of viruses, these domains are present in important viral proteins like helicases, nucleases, polymerases, capsid proteins or regulation factors. Therefore, they are implicated in the replicative cycle and parasitic processes of viruses. That is why it is possible to think that those domains present low levels of divergence due to selective pressures. For these reasons, the main goal for this project is to create a catalogue of the RNA-binding domains found in all the available viral proteomes, using bioinformatics tools in order to analyze its evolutionary process, and thus shed light on the general virus evolution. ProDom database was used to obtain larger than six thousand RNA-binding domain families that belong to the three cellular domains of life and some viral groups. From the sequences of these families, protein profiles were created using HMMER 3.1 tools in order to find distant homologous within greater than four thousand viral proteomes available in GenBank. Once accomplished the analysis, almost three thousand hits were obtained in the viral proteomes. The homologous sequences were found in proteomes of the principal Baltimore viral groups, showing interesting distribution patterns that can contribute to understand the evolution of viruses and their host-virus interactions. Presence of cellular RNA-binding domains within virus proteomes seem to be explained by closed interactions between viruses and their hosts. Recruitment of these domains is advantageous for the viral fitness, allowing viruses to be adapted to the host cellular environment.

Keywords: bioinformatics tools, distant homology, RNA-binding domains, viral evolution

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Authors: Mona Marzouk, Nesrine Hegazi, Aliaa Ragheb, Mona El Shabrawy, Salwa Kawashty

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The whole plant of Matthiola longipetala (Brassicaceae) was extracted by 70% methanol to give the total aqueous methanol extract (AME), which was defatted by hexane yielded hexane extract (HE) and defatted AME (DAME). HE was analyzed through GC/MS assay and revealed the detection of 28 non-polar compounds. In addition, the chemical investigation of DAME led to the isolation and purification of twelve flavonoids and three chlorogenic acids. Their structures were interpreted through chemical (complete and partial acid hydrolysis) and spectroscopic analysis (MS, UV, 1D and 2D NMR). Among them, nine compounds have been isolated for the first time from M. longipetala. Moreover, LC-ESI-MS analysis of DAME was achieved to detect additional 46 metabolites, including phospholipids, organic acids, phenolic acids and flavonoids. The biological activity of AME, HE and DAME against alloxan inducing oxidative stress and diabetes in male rats was investigated. Diabetes was induced using a single dose of Alloxan (150 mg/kg b.wt.). HE and DAME significantly increased serum GSH content in rats (37.3±0.7 and 35.9±0.6 mmol/l) compared to diabetic rats (21.8±0.3) and vitamin E (36.2±1.1) at P<0.01. Also, HE, DAME and AME revealed a significant acute anti-hyperglycemic effect potentiated after four weeks of treatment with blood glucose levels of 96.2±5.4, 98.7±6.1 and 98.9±8.6 mg/dl, respectively, compared to diabetic rats (263.4±7.8) and metaformin group (81.9±2.4) at P<0.01.

Keywords: Brassicaceae, Flavonoid, LCMS/MS, Matthiola

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Authors: Lebea Nthunya, Arne Verliefde, Bhekie Mamba, Sabelo Mhlanga

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A study aimed at developing membrane distillation (MD) processes that can be used for brackish/saline water purification will be presented. MD is a membrane-based technology that presents a possibility to counteract challenges associated with pressure driven membranes at high separation efficiencies. Membrane distillation membranes (MDM) are affected by wettability and fouling. Wetting inside the pores of the membrane is elevated by the hydrophilic characteristic of the membrane, while fouling is mostly induced by the hydrophobic-hydrophobic interaction of pollutants and the surface of the hydrophobic membranes, hence block the pores of the membranes. These properties are not desirable. As such, a carefully designed polyvinylidene fluoride (PVDF) MDM composed of a super-hydrophobic modified backbone and a super-hydrophilic thin layer has been developed to concurrently overcome these challenges. The membranes were characterized using contact angle measurements to confirm their hydrophobicity/hydrophilicity. SEM and SAXS were used to study the morphology and pore distribution on the surface of the membrane. The contact angles of the active surface ≤ 30º and that of the backbone ≥ 140º has thus revealed that the active surface was highly hydrophilic while the backbone was highly hydrophobic. The SEM and the SAXS results have also confirmed that the membranes are highly porous. These materials demonstrated a potential to remove salts from water at high efficiencies.

Keywords: membrane distillation, modification, energy efficiency, desalination

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Authors: Nusrat Praween, Krishna Thej Pammi Guru, Palash Kumar Basu

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Designing non-invasive biosensors for cancer diagnosis is essential for developing an affordable and specific tool to measure cancer-related exosome biomarkers. Exosomes, released by healthy as well as cancer cells, contain valuable information about the biomarkers of various diseases, including cancer. Despite the availability of various isolation techniques, ultracentrifugation is the standard technique that is being employed. Post isolation, exosomes are traditionally exposed to detergents for extracting their proteins, which can often lead to protein degradation. Further to this, it is very essential to develop a sensing platform for the quantification of clinically relevant proteins in a wider range to ensure practicality. In this study, exosomes were immobilized on the Au Screen Printed Electrode (SPE) using EDC/NHS chemistry to facilitate binding. After immobilizing the exosomes on the screen-printed electrode (SPE), we investigated the impact of the electric field by applying various voltages to induce exosome lysis and release their contents. The lysed solution was used for sensing TSG101, a crucial biomarker associated with various cancers, using both faradaic and non-faradaic electrochemical impedance spectroscopy (EIS) methods. The results of non-faradaic and faradaic EIS were comparable and showed good consistency, indicating that non-faradaic sensing can be a reliable alternative. Hence, the non-faradaic sensing technique was used for label-free quantification of the TSG101 biomarker. The results were validated using ELISA. Our electrochemical immunosensor demonstrated a consistent response of TSG101 from 125 pg/mL to 8000 pg/mL, with a detection limit of 0.125 pg/mL at room temperature. Additionally, since non-faradic sensing is label-free, the ease of usage and cost of the final sensor developed can be reduced. The proposed immunosensor is capable of detecting the TSG101 protein at low levels in healthy serum with good sensitivity and specificity, making it a promising platform for biomarker detection.

Keywords: biosensor, exosomes isolation on SPE, electric field lysis of exosome, EIS sensing of TSG101

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Authors: Amany Ragab, Nashwa Khairat Abousamra, Omayma Saleh, Asmaa Higazy

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Insulin resistance syndrome has been shown to be associated with many coagulation and fibrinolytic proteins and these associations suggest that some coagulation and fibrinolytic proteins have a role in atherothrombotic disorders. This study was conducted to determine the levels of some of the haemostatic parameters in subjects having metabolic syndrome and to correlate these values with the anthropometric and metabolic variables associated with this syndrome. The study included 46 obese non diabetic subjects of whom 28 subjects(group1) fulfilled the ATP III criteria of the metabolic syndrome and 18 subjects (group2) did not have metabolic syndrome as well as 14 lean subjects (group 3) of matched age and sex as a control group. Clinical and laboratory evaluation of the study groups stressed on anthropometric measurements (weight, height, body mass index, waist circumference, and sagittal abdominal diameter), blood pressure, and laboratory measurements of fasting plasma glucose, fasting insulin, serum lipids, tissue plasminogen activator (t-PA), antithrombin III activity (ATIII), protein C and von Willebrand factor (vWf) antigen. There was significant increase in the concentrations of t-PA and vWf antigens in subjects having metabolic syndrome (group 1) in comparison to the other groups while there were non-significant changes in the levels of protein C antigen and AT III activity. Both t-PA and vWf showed significant correlation with HOMA-IR as a measure of insulin sensitivity. The t-PA showed also significant correlation with most of the variables of metabolic syndrome including waist circumference, BMI, systolic blood pressure, fasting plasma glucose, fasting insulin, and HDL cholesterol. On the other hand, vWf showed significant correlations with fasting plasma glucose, fasting insulin and sagital abdominal diameter, with non-significant correlations with the other variables. Haemostatic and fibrinolytic parameters should be included in the features and characterization of the insulin resistance syndrome. t-PA and vWf antigens concentrations were increased in subjects with metabolic syndrome and correlated with the HOMA-IR measure of insulin sensitivity. Taking into consideration that both t-PA and vWf are mainly released from vascular endothelium, these findings could be an indicator of endothelial dysfunction in that group of subjects.

Keywords: insulin resistance, obesity, metabolic syndrome, coagulation

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Authors: Mulugeta Gurum Gerechal

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Nowadays, the photocatalytic mechanism of water purification using nanoparticles has gained wider acceptance. For this purpose, the Crystal form of N- TiO₂ and Ag-TiO₂ was prepared from TiCl₄, Urea, NH₄OH and AgNO₃ by sol-gel method and simple solid phase reaction followed by calcination at a temperature of 400 °C for 4h at each. The synthesized photocatalysts were characterized using XRD, SEM and UV-visible diffuse reflectance spectra. In the experiment, it was found that the absorption edge of N-TiO₂ was a well efficient shift to visible light as compared to Ag-TiO₂. The XRD diffraction makes the particle size of N-TiO₂ smaller than Ag-TiO₂. The effect of catalyst loading and the effect of temperature on the photocatalytic efficiency of the prepared samples was tested using methylene blue as a target pollutant. The photocatalytic degradation efficiency of the catalysts for methylene blue was increased from 57.05 to 96.02% under solar radiation as the amount of the catalyst increased from 0.15 to 0.45 gram for N-TiO₂. Similarly, photocatalytic degradation of methylene blue was increased from 40.32 to 81.21% as the amount of Ag-TiO₂ increased from 0.05g to 0.1g. In addition, the photocatalytic degradation efficiency of the catalysts for the removal of methylene blue was increased from 58.00 to 98.00 and 47.00 to 81.21 % under solar radiation as the calcination temperature of the catalyst increased from 300 to 500 for N-TiO₂ for Ag-TiO₂ 300 to 4000C. However, a further increase in catalyst loading and calcination temperature was found to decrease the degradation efficiency.

Keywords: photocatalysis, degradation, nanoparticles, catalyst loading, calcination and methylene blue

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Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

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Authors: A. Gilewska, J. Masternak, K. Kazimierczuk, L. Turlej, J. Wietrzyk, B. Barszcz

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Platinum-based drugs are now widely used as chemotherapeutic agents. However the platinum complexes show the toxic side-effects: i) the development of platinum resistance; ii) the occurrence of severe side effects, such as nephro-, neuro- and ototoxicity; iii) the high toxicity towards human fibroblast. Therefore the development of new anticancer drugs containing different transition-metal ions, for example, ruthenium, rhodium, iridium is a valid strategy in cancer treatment. In this paper, we reported the synthesis, spectroscopic, structural and biological properties of complexes of ruthenium, rhodium, and iridium containing N,N-chelating ligand (2,2’-bisimidazole). These complexes were characterized by elemental analysis, UV-Vis and IR spectroscopy, X-ray diffraction analysis. These complexes exhibit a typical pseudotetrahedral three-legged piano-stool geometry, in which the aromatic arene ring forms the seat of the piano-stool, while the bidentate 2,2’-bisimidazole (ligand) and the one chlorido ligand form the three legs of the stool. The spectroscopy data (IR, UV-Vis) and elemental analysis correlate very well with molecular structures. Moreover, the cytotoxic activity of the complexes was carried out on human cancer cell lines: LoVo (colorectal adenoma), MV-4-11 (myelomonocytic leukaemia), MCF-7 (breast adenocarcinoma) and normal healthy mouse fibroblast BALB/3T3 cell lines. To predict a binding mode, a potential interaction of metal complexes with calf thymus DNA (CT-DNA) and protein (BSA) has been explored using UV absorption and circular dichroism (CD). It is interesting to note that the investigated complexes show no cytotoxic effect towards the normal BALB/3T3 cell line, compared to cisplatin, which IC₅₀ values was determined as 2.20 µM. Importantly, Ru(II) displayed the highest activity against HL-60 (IC₅₀ 4.35 µM). The biological studies (UV-Vis and circular dichroism) suggest that arene-complexes could interact with calf thymus DNA probably via an outside binding mode and interact with protein (BSA).

Keywords: ruthenium(II) complex, rhodium(III) complex, iridium(III) complex, biological activity

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Authors: Misty Attwood, Helgi Schioth

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Transmembrane proteins are important components in many essential cell processes such as signal transduction, cell-cell signalling, transport of solutes, structural adhesion activities, and protein trafficking. Due to their involvement in diverse critical activities, transmembrane proteins are implicated in different disease pathways and hence are the focus of intense interest in understanding functional activities, their pathogenesis in disease, and their potential as pharmaceutical targets. Further, as the structure and function of proteins are correlated, investigating a group of proteins with the same tertiary structure, i.e., the same number of transmembrane regions, may give understanding about their functional roles and potential as therapeutic targets. In this in silico bioinformatics analysis, we identify and comprehensively characterize the previously unstudied group of proteins with five transmembrane-spanning regions (5TM). We classify nearly 60 5TM proteins in which 31 are members of ten families that contain two or more family members and all members are predicted to contain the 5TM architecture. Furthermore, nine singlet proteins that contain the 5TM architecture without paralogues detected in humans were also identifying, indicating the evolution of single unique proteins with the 5TM structure. Interestingly, more than half of these proteins function in localization activities through movement or tethering of cell components and more than one-third are involved in transport activities, particularly in the mitochondria. Surprisingly, no receptor activity was identified within this family in sharp contrast with other TM families. Three major 5TM families were identified and include the Tweety family, which are pore-forming subunits of the swelling-dependent volume regulated anion channel in astrocytes; the sidoreflexin family that acts as mitochondrial amino acid transporters; and the Yip1 domain family engaged in vesicle budding and intra-Golgi transport. About 30% of the proteins have enhanced expression in the brain, liver, or testis. Importantly, 60% of these proteins are identified as cancer prognostic markers, where they are associated with clinical outcomes of various tumour types, indicating further investigation into the function and expression of these proteins is important. This study provides the first comprehensive analysis of proteins with 5TM regions and provides details of the unique characteristics and application in pharmaceutical development.

Keywords: 5TM, cancer prognostic marker, drug targets, transmembrane protein

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Authors: Vaibhav D. Bhatt, Anju P. Kunjadia, D. S. Nauriyal, Bhumika J. Joshi, Chaitanya G. Joshi

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Metagenomic analysis of milk samples collected from local cattle breed, kankrej (Bos indicus), Gir (Bos indicus) and Crossbred (Bos indicus X Bos taurus) cattle harbouring subclinical mastitis was carried out by next-generation sequencing (NGS) 454 GS-FLX technology. Around 56 different species including members of Enterobacteriales, Pseudomonadales, Bacillales and Lactobacillales with varying abundance were detected in infected milk. The interesting presence of bacteriophages against Staphylococcus aureus, Escherichia coli, Enterobacter and Yersinia species were observed, especially Enterobacteria and E. coli phages (0∙32%) in Kankrej, Enterobacteria and Staphylococcus phages (1∙05%) in Gir and Staphylococcus phages (2∙32%) in crossbred cattle. NGS findings suggest that phages may be involved in imparting natural resistance of the cattle against pathogens. Further infected milk samples were subjected for bacterial isolation. Fourteen different isolates were identified, and DNA was extracted. Genes (Tet-K, Msr-A, and Mec-A) providing antibiotic resistance to the bacteria were screened by Polymerase Chain Reaction and results were validated with traditional antibiotic assay. Total 3 bacteriophages were isolated from nearby environment of the cattle farm. The efficacy of phages was checked against multi-drug resistant bacteria, identified by PCR. In-vivo study was carried out for phage therapy in mammary glands of female rats “Wister albino”. Mammary glands were infused with MDR isolates for 3 consecutive days. Recovery was observed in infected rats after intramammary infusion of sterile phage suspension. From day 4th onwards, level of C-reactive protein was significant increases up to day 12th . However, significant reduction was observed between days 12th to 18th post treatment. Bacteriophages have significant potential as antibacterial agents and their ability to replicate exponentially within their hosts and their specificity, make them ideal candidates for more sustainable mastitis control.

Keywords: bacteriophages, c-reactive protein, mastitis, metagenomic analysis

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Authors: F. Alaa Ali, S. A. Sherein Abdelgayed, S. Osama. EL-Tawil, M. Adel Bakeer

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Tartrazine is an organic azo dyes food additive widely used in foods, drugs, and cosmetics. The present study aimed to investigate the toxic effects of tartrazine on kidneys and liver biomarkers in addition to the investigation of oxidative stress and change of histopathological structure of liver and kidneys in 30 male rats. Tartrazine was orally administrated daily at dose 200 mg/ kg bw (1/ 10 LD₅₀) for sixty days. Serum and tissue samples were collected at the end of the experiment to investigate the underlying mechanism of tartrazine through assessment oxidative stress (Glutathione (GSH), Superoxide dismutase (SOD) and malondialdehyde (MDA) and biochemical markers (alanine aminotransferase (ALT), aspartate aminotransferase (AST), Total protein and Urea). Liver and kidneys tissue were collected and preserved in 10% formalin for histopathological examination. The obtained values were statistically analyzed by one way analysis of variance (ANOVA) followed by multiple comparison test. Biochemical analysis revealed that tartrazine induced significant increase in serum ALT, AST, total protein, urea level compared to control group. Tartrazine showed significant decrease in liver GSH and SOD where their values when compared to control group. Tartrazine induced increase in liver MDA compared to control group. Histopathology of the liver showed diffuse vacuolar degeneration in hepatic parenchyma, the portal area showed sever changes sever in hepatoportal blood vessels and in the bile ducts. The kidneys showed degenerated tubules at the cortex together with mononuclear leucocytes inflammatory cells infiltration. There is perivascular edema with inflammatory cell infiltration surrounding the congested and hyalinized vascular wall of blood vessel. The present study indicates that the subchronic effects of tartrazine have a toxic effect on the liver and kidneys together with induction of oxidative stress by formation of free radicals. Therefore, people should avoid the hazards of consuming tartrazine.

Keywords: albino rats, tartrazine, toxicity, pathology

Procedia PDF Downloads 359

Authors: Anna Martin, Raffael Osen

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The demand for fish from aquaculture is constantly growing. Concurrently, due to a shortage of fishmeal caused by extensive overfishing, fishmeal substitution by plant proteins is getting increasingly important for the production of sustainable aquafeed. Several research studies evaluated the impact of plant protein meals, concentrates or isolates on fish health and fish feed quality. However, these protein raw materials often require elaborate and expensive manufacturing and their availability is limited. Rapeseed press cake (RPC) – a side product of de-oiling processes – exhibits a high potential as a plant-based fishmeal alternative in fish feed for carnivorous species due to its availability, low costs and protein content. In order to produce aquafeed with RPC, it is important to systematically assess i) inclusion levels of RPC with similar pellet qualities compared to fishmeal containing formulations and ii) how extrusion parameters can be adjusted to achieve targeted pellet qualities. However, the effect of RPC on extrusion system parameters and pellet quality has only scarcely been investigated. Therefore, the aim of this study was to evaluate the impact of feed formulation, extruder barrel temperature (90, 100, 110 °C) and screw speed (200, 300, 400 rpm) on extrusion system parameters and the physical properties of fish feed pellets. A co-rotating pilot-scale twin screw extruder was used to produce five iso-nitrogenous feed formulations: a fish meal based reference formulation including 16 g/100g fishmeal and four formulations in which fishmeal was substituted by RPC to 25, 50, 75 or 100 %. Extrusion system parameters, being product temperature, pressure at the die, specific mechanical energy (SME) and torque, were monitored while samples were taken. After drying, pellets were analyzed regarding to optical appearance, sectional and longitudinal expansion, sinking velocity, bulk density, water stability, durability and specific hardness. In our study, the addition of minor amounts of RPC already had high impact on pellet quality parameters, especially on expansion but only marginally affected extrusion system parameters. Increasing amounts of RPC reduced sectional expansion, sinking velocity, bulk density and specific hardness and increased longitudinal expansion compared to a reference formulation without RPC. Water stability and durability were almost not affected by RPC addition. Moreover, pellets with rapeseed components showed a more coarse structure than pellets containing only fishmeal. When the adjustment of barrel temperature and screw speed was investigated, it could be seen that the increase of extruder barrel temperature led to a slight decrease of SME and die pressure and an increased sectional expansion of the reference pellets but did almost not affect rapeseed containing fish feed pellets. Also changes in screw speed had little effects on the physical properties of pellets however with raised screw speed the SME and the product temperature increased. In summary, a one-to-one substitution of fishmeal with RPC without the adjustment of extrusion process parameters does not result in fish feed of a designated quality. Therefore, a deeper knowledge of raw materials and their behavior under thermal and mechanical stresses as applied during extrusion is required.

Keywords: extrusion, fish feed, press cake, rapeseed

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Authors: Nematollah Gheibi, Nader Divan Khosroshahi, Mahdi Mohammadi Ghanbarlou

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Introduction: Nowadays, the phenomenon of bacterial resistance is one of the most important challenge of the health community in the world. Propolis is most important production of bee colonies that collected from of various plants. So far, a lot of investigations carried out about its antibacterial effects. Material and methods: Thirty gram of propolis prepared as ethanolic extract and after different process of purification, 7.5 gr of its pure form were obtained. Propolis compounds identification was performed by TLC and VLC methods. The HPLC spectrum obtaining from propolis ethanolic extract was compared with some purified standard phenolic and flavonoid substances. Antibacterial effects of ethanol extract of purified propolis were evaluated on two strains of Staphylococcus aureus and Pseudomonas aeruginosa and their MIC was determined by the microdillution assay. Results: Ethanolic propolis extraction analyzed by TLC were resulted to confirm several phenolic and flavonoid compounds in this extract and some of the confirmed by HPLC technique. Minimum inhibitory concentration (MIC) for standard Staphylococcus aureus (ATCC25923) and Pseudomonas aeruginosa (ATCC27853) strains were obtained 2.5 mg/ml and 50 mg/ml respectively. Conclusion: Bee Propolis is a mix organic compound that has a lot of beneficial effects such as anti-bacterial that emphasized in this investigation. It is proposed as a rich source of natural phenolic and flavonoids compounds in designing of new biological resources for hygienic and medical applications.

Keywords: propolis, Staphylococcus aureus, Pseudomonas aeruginosa, antibacterial

Procedia PDF Downloads 307

Authors: Mei-Ling Chan, Jin-Ming Wu, Konstantin V. Kudryavtsev, Jih-Hwa Guh

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Prostate cancer is one of the most common malignant disease in men. KUD983 is an enantiomerically pure β-dipeptide derivative, which may have anti-cancer effects. In the present study, KUD983 exhibits powerful activity against hormone-refractory prostate cancer (HRPC) PC-3 and DU145 cells. The IC50 values of KUD983 in PC-3 and DU145 cells are 0.56±0.07M and 0.50±0.04 M respectively. KUD983 induced G1 arrest of the cell cycle and subsequent apoptosis associated with the down-regulation of several related proteins including cyclin D1, cyclin E and Cdk4, and the de-phosphorylation of RB. The protein expressions of nuclear and total c-Myc protein, which was able to regulate the expression of both cyclin D1 and cyclin E, were significantly suppressed by KUD983. Phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) is an important signaling pathway that influences the energy metabolism, cell cycle, proliferation, survival and apoptosis of cells, and is associated with numerous other signaling pathways. The Western Blot data revealed that KUD983 inhibited PI3K/Akt and mTOR/p70S6K/4E-BP1 pathways. The transient transfection of constitutively active myristylated Akt (myr-Akt) cDNA significantly reversed KUD983-induced caspase activation but did not abolish the suppression of mTOR/p70S6K/4E-BP1 signaling cascade indicating the presence of both Akt-dependent and -independent pathways. Moreover, KUD983-induced effect was collaborated with the down-regulation of anti-apoptotic Bcl-2 members (e.g., Bcl-2, and Mcl-1) and IAP family members (e.g., survivin). Furthermore, KUD983 induced autophagic cell death using confocal microscopic examination, investigating the level of conversion of LC3-I to LC3-II and flow cytometric detection of AVO-positive cells. Taken together, the data suggest that KUD983 is an anticancer β-dipeptide against HRPCs through the inhibition of cell proliferation and induction of apoptotic and autophagic cell death. The suppression of signaling pathways mediated by c-Myc, PI3K/Akt and mTOR/p70S6K/4E-BP1 and the collaboration with down-regulation of Mcl-1 and survivin may indicate the mechanism of KUD983 against HRPC.

Keywords: β-dipeptide, hormone-refractory prostate cancer, mTOR, PI3K/Akt

Procedia PDF Downloads 282

Authors: Chona Gelani, Mylene Uy, Keisuke Yasuda, Emi Ohta, Shinji Ohta

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The marine environment is undoubtedly a rich source of diverse organisms that possess bioactive secondary metabolites with important pharmacological activities. Marine sponges have since been contributing a wide array of compounds of biomedical and pharmaceutical importance. This study is an attempt to contribute to the growing and advancing marine natural products research. It aims to evaluate the cytotoxicity of the hexane extract (H) from the Philippine marine sponges, Rhabdastrella globostellata (Rg), Callyspongia sp. (Calsp), Callyspongia aerizusa (Ca), Carteriospongia sp. (Carsp), and Cinachyrella sp. (Cisp) using the eggs of starfish, Asterina pectinifera, and sea urchin, Hemicentrotus pulcherrimus. Specifically, the cytotoxicity of the marine sponge hexane extract was determined through its inhibition of starfish and sea urchin embryonic development. After 24 hours, CarspH and RgH inhibited early gastrulation of sea urchin at a minimum concentration of 15.63 and 31.25 μg/mL, respectively. CalspH inhibited the early gastrulation of both sea urchin and starfish at 125 μg/mL, whereas CaH halted the morula of sea urchin and early gastrulation of starfish at 250 μg/mL. CispH exhibited relatively weak inhibitory activity on starfish embryogenesis but inhibited the early gastrulation of sea urchin at 250 μg/mL. The results obtained from this study were used as basis for the separation, isolation and purification of the component(s) of the hexane extracts from the five Philippine marine sponges.

Keywords: embryonic development, marine sponge cytotoxicity, Philippine marine sponges, sea urchin and starfish embryogenesis

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Authors: O. J. Osunkeye, P. O. Fakolade, B. E. Olorede

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Broilers productions depend on the provision of adequate and goo quality feed containing all the nutrients, including proteins, carbohydrate, fats, vitamins, minerals and water. All these nutrients have to be provided at a required amount to support maximum productivity and normal physiological functions and demands. Among these nutrients proteins are particularly important, since they are essential for meat and muscle production, optimum growth and health status. Poultry production industry in the developing countries is been threatened because of the over dependency on Soybean meal as one of the key/major conventional protein stuff for feeding livestock. Even the competition between man and livestock for Soybean and other protein sources made the price of this feed stuff to be on the increase. Hence the needs to seek for an alternative feed stuff which is cheap and less competitive. This study showed the blood profile, organ and carcass characteristics and performance of broilers fed with Cowpea Testa Meal (CTM) based diets. Four diets were formulated with Cowpea Testa replacing Soybean at 0%, 15%, 30%, and 50% graded levels. One hundred and twenty day-old unsexed broiler birds were allotted to these four treatments with 3 replicates of 10 birds per replicate. The results showed no significant differences in all the haematological parameters measured (P>0.05), the serum metabolites analysis revealed significant different in Cholesterol (99.8 mg/dl, 112.84 mg/dl, 131.07 mg/dl and 97.66 mg/dl respectively) (P<0.05) among others. There were significant differences within the diets for average daily weight gain, average feed intake and feed to gain ratio. The birds on control (0%) and CTM gained more weight than those fed with 30% and 50% CTM diets. The organs and carcass primal cuts of the broilers expressed significant different for the spleen (0.12 g, 0.09 g, 0.11 g and 0.14 g respectively), lungs (0.97 g, 0.72 g, 0.77 g and 1.01g respectively) and proventriculus (0.96 g, 0.99 g, 0.81 g and 0.85 g respectively) (P<0.05). For the carcass, there were no significant differences (P<0.05) in the breast, thigh, drumstick, wing and neck except for the Back (21.27 g, 21.04 g, 17.71 g, and 17.89 g respectively). In conclusion, CTM inclusion in broiler’s diet could be used as an alternative feed stuff in replacement of Soybean meal up to 15% without any adverse effects as revealed by the blood profile and to increase the growth performance of the birds.

Keywords: physiological functions, cholesterol, blood profiles, CTM and carcass analysis

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Authors: I. A. Yola

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Impact of poultry dropping, cow dung and rumen content on utilization and proximate composition of Oreochromis niliticus, Clarias gariepinus and Cyprinus capio in a polyculture system were studied. The research was conducted over a period of 52 weeks. Poultry droppings (PD), cow dung (CD) and rumen content (RC) were applied at three levels 30g,60g and 120g/m2/week, 25g,50g and 100g/m2/week and 22g, 44g and 88g/m2/week treatment, respectively. The control only conventional feed with 40% CP without manure application was used. Physicochemical and biological properties measured were higher in manure pond than control. The difference was statistically significant (P < 0.05) between and within treatments with exception of temperature with a combined mean of 27.900C. The water was consistently alkaline with mean values for pH of 6.61, transparency 22.6cm, conductivity 35.00µhos/cm, dissolved oxygen 4.6 mg/l, biological oxygen demand 2.8mg/l, nitrate and phosphates 0.9mg/l and 0.35mg/l, respectively. The three fish species increase in weight with increased manure rate, with a higher value in PD treatment on C. capio record 340g, O. niloticus weighed 310g and C. gariepinus 280g over the experimental period. Fishes fed supplementary diet (control) grew bigger with highest value on C. capio (685g) O. niloticus (620g) and then C. gariepinus (526g). The differences were statistically significant (P < 0.05). The result of whole body proximate analysis indicated that various manures and rates had an irregular pattern on the protein and ash gain per 100g of fish body weight gain. The combined means for whole fish carcass protein, lipids, moisture, ash and gross energy were 11.84, 2.43, 74.63, 3.00 and 109.9 respectively. The notable exceptions were significant (p < 0.05) increases in body fat and gross energy gains in all fish species accompanied by decreases in percentages of moisture as manure rates increased. Survival percentage decreases from 80% to 70%. It is recommended to use poultry dropping as manure/feeds at the rate of 120kg/ha/week for good performances in polyculture.

Keywords: organic manure, Nile tilapia, African mud fish, common carp, proximate composition

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Authors: Yu Yoshihara, Chika Tada, Moe Takada, Nyam-Osor Purevdorj, Khorolmaa Chimedtseren, Yutaka Nakai

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Water pollution from animal waste and its influence on grazing animals is a current concern regarding Mongolian grazing lands. We allocated 32 free-grazing lambs to four groups and provided each with water from a different source (upper stream, lower stream, well, and pond) for 49 days. We recorded the amount of water consumed by the lambs, as well as their body weight, behavior, white blood cell count, acute phase (haptoglobin) protein level, and fecal condition. We measured the chemical and biological qualities of the four types of water, and we detected enteropathogenic and enterohemorrhagic Escherichia coli in fecal samples by using a genetic approach. Pond water contained high levels of nitrogen and minerals, and well water contained high levels of bacteria. The odor concentration index decreased in order from pond water to upper stream, lower stream, and well. On day 15 of the experiment, the following parameters were the highest in lambs drinking water from the following sources: water intake (pond or lower stream), body weight gain (pond), WBC count (lower stream), haptoglobin concentration (well), and enteropathogenic E. coli infection rate (lower stream). Lambs that drank well water spent more time lying down and less time grazing than the others, and lambs that drank pond water spent more time standing and less time lying down. Lambs given upper or lower stream water exhibited more severe diarrhea on day 15 of the experiment than before the experiment. Mongolian sheep seemed to adapt to chemically contaminated water: their productivity benefited the most from pond water, likely owing to its rich mineral content. Lambs that drank lower stream water showed increases in enteropathogenic E. coli infection, clinical diarrhea, and WBC count. Lambs that drank well water, which was bacteriologically contaminated, had increased serum acute phase protein levels and poor physical condition; they were thus at increased risk of negative health and production effects.

Keywords: DNA, Escherichia coli, fecal sample, lower stream, well water

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