Search results for: animal protein (AP)

Authors: O. Ubani, H. I. Atagana, M. S. Thantsha, R. Adeleke

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The oil sludge components (polycyclic aromatic hydrocarbons, PAHs) have been found to be cytotoxic, mutagenic and potentially carcinogenic and microorganisms such as bacteria and fungi can degrade the oil sludge to less toxic compounds such as carbon dioxide, water and salts. In the present study, we isolated different bacteria with PAH-degrading potentials from the co-composting of oil sludge and different animal manure. These bacteria were isolated on the mineral base medium and mineral salt agar plates as a growth control. A total of 31 morphologically distinct isolates were carefully selected from 5 different compost treatments for identification using polymerase chain reaction (PCR) of the 16S rDNA gene with specific primers (16S-P1 PCR and 16S-P2 PCR). The amplicons were sequenced and sequences were compared with the known nucleotides from the gene bank database. The phylogenetical analyses of the isolates showed that they belong to 3 different clades namely Firmicutes, Proteobacteria and Actinobacteria. These bacteria identified were closely related to genera Bacillus, Arthrobacter, Staphylococcus, Brevibacterium, Variovorax, Paenibacillus, Ralstonia and Geobacillus species. The results showed that Bacillus species were more dominant in all treated compost piles. Based on their characteristics these bacterial isolates have high potential to utilise PAHs of different molecular weights as carbon and energy sources. These identified bacteria are of special significance in their capacity to emulsify the PAHs and their ability to utilize them. Thus, they could be potentially useful for bioremediation of oil sludge and composting processes.

Keywords: bioaugmentation, biodegradation, bioremediation, composting, oil sludge, PAHs, animal manures

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Authors: Fana Woldetsadik

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Ethiopia has a huge livestock population, including sheep, that has been contributing a considerable portion to the economy of the country and still promising to rally around the economic advancement of the country. However, feed shortage is a limiting factor in the production and productivity of sheep among Ethiopian smallholder farmers. Therefore, the aim of this study was to prove the role of the locally available brewery by-products called dried Atella as a supplement in feed intake, digestibility, live weight gain, carcass yield, and economic benefit in comparison with commercially purchased supplements known as niger seed cake (NSC). This on-station feeding experiment was conducted on the Zenzelma Campus of Bahir Dar University animal farm. The experimental design used for this research was a completely randomized design (CRD) with five replications. The crude protein (CP) content of dried Atella, wheat bran (WB), natural pasture hay (NPH) and NSC were about 25.07%, 16.57%, 4.48% and 38.04%, respectively, while the neutral detergent fibre (NDF),acid detergent fibre (ADF) and acid detergent lignin (ADL) content of dried Atella, WB, NPH and NSC were around 31.75%, 8.31%, 8.14%; 42.05%, 22.64%, 4.04%; 74.21%, 50.81%, 8.66%; 42.31%, 26.95% and 6.9%, respectively. The result depicted that a higher(P < 0.001) feed intake, nutrient intake, and digestibility for lambs supplemented with Atella than those supplemented with NSC. Furthermore, daily body weight gain and carcass characteristics were better (P < 0.05) for the sheep supplemented with dried Atella than NSC. On the other hand, in terms of profitability, although there was no substantial difference (P > 0.05) between T2 (animals fed NPH,NSC and WB) and T3 (animals fed NPH, Atella and WB), slightly better benefit was recorded in T3 groups. However, loss of money was recorded in T1 (animals fed NPH and WB). Hence, from the biological performance of lambs, it was concluded that Atella could be a potential supplementary feed for sheep fattening among smallholder farmers than NSC despite no profitability difference. Nevertheless, further investigation is recommended to examine the consequence of supplementation of NPH with NSC and NPH with Atella on fatty acid profile analysis, the physicochemical composition of meat, and meat composition.

Keywords: Attela, Bahir Dar university, Carcass yield, digestibility, natural pasture hay, Niger seed cake, smallholder farmers, weight gain, Ethiopia

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Authors: Danna Jia, Bin Li

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Background: Atopic dermatitis (AD) is a chronic and refractory inflammatory skin disease characterized by relapsing eczematous and pruritic skin lesions. The global prevalence of AD ranges from 1~ 20%, and its incidence rates are increasing. It affects individuals from infancy to adulthood, significantly impacting their daily lives and social activities. Despite its major health burden, the precise mechanisms underlying AD remain unknown. Understanding the genetic differences associated with AD is crucial for advancing diagnosis and targeted treatment development. This study aims to identify candidate genes of AD by using bioinformatics analysis. Methods: We conducted a comprehensive analysis of four pooled transcriptomic datasets (GSE16161, GSE32924, GSE130588, and GSE120721) obtained from the Gene Expression Omnibus (GEO) database. Differential gene expression analysis was performed using the R statistical language. The differentially expressed genes (DEGs) between AD patients and normal individuals were functionally analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Furthermore, a protein-protein interaction (PPI) network was constructed to identify candidate genes. Results: Among the patient-level gene expression datasets, we identified 114 shared DEGs, consisting of 53 upregulated genes and 61 downregulated genes. Functional analysis using GO and KEGG revealed that the DEGs were mainly associated with the negative regulation of transcription from RNA polymerase II promoter, membrane-related functions, protein binding, and the Human papillomavirus infection pathway. Through the PPI network analysis, we identified eight core genes: CD44, STAT1, HMMR, AURKA, MKI67, and SMARCA4. Conclusion: This study elucidates key genes associated with AD, providing potential targets for diagnosis and treatment. The identified genes have the potential to contribute to the understanding and management of AD. The bioinformatics analysis conducted in this study offers new insights and directions for further research on AD. Future studies can focus on validating the functional roles of these genes and exploring their therapeutic potential in AD. While these findings will require further verification as achieved with experiments involving in vivo and in vitro models, these results provided some initial insights into dysfunctional inflammatory and immune responses associated with AD. Such information offers the potential to develop novel therapeutic targets for use in preventing and treating AD.

Keywords: atopic dermatitis, bioinformatics, biomarkers, genes

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Authors: B. Cilek Tatar, G. Sumnu, M. Oztop, E. Ayaz

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Encapsulation protects sensitive food ingredients against heat, oxygen, moisture and pH until they are released to the system. It can mask the unwanted taste of nutrients that are added to the foods for fortification purposes. Cherry laurels (Prunus laurocerasus) contain phenolic compounds which decrease the proneness to several chronic diseases such as types of cancer and cardiovascular diseases. The objective of this research was to study the effects of centrifugation, different coating materials and homogenization methods on microencapsulation of powders obtained from cherry laurels. In this study, maltodextrin and mixture of maltodextrin:whey protein with a ratio of 1:3 (w/w) were chosen as coating materials. Total solid content of coating materials was kept constant as 10% (w/w). Capsules were obtained from powders of freeze-dried cherry laurels through encapsulation process by silent crusher homogenizer or microfluidization. Freeze-dried cherry laurels were core materials and core to coating ratio was chosen as 1:10 by weight. To homogenize the mixture, high speed homogenizer was used at 4000 rpm for 5 min. Then, silent crusher or microfluidizer was used to complete encapsulation process. The mixtures were treated either by silent crusher for 1 min at 75000 rpm or microfluidizer at 50 MPa for 3 passes. Freeze drying for 48 hours was applied to emulsions to obtain capsules in powder form. After these steps, dry capsules were grounded manually into a fine powder. The microcapsules were analyzed for total antioxidant activity with DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method. Prior to high speed homogenization, the samples were centrifuged (4000 rpm, 1 min). Centrifugation was found to have positive effect on total antioxidant activity of capsules. Microcapsules treated by microfluidizer were found to have higher total antioxidant activities than those treated by silent crusher. It was found that increasing whey protein concentration in coating material (using maltodextrin:whey protein 1:3 mixture) had positive effect on total antioxidant activity for both silent crusher and microfluidization methods. Therefore, capsules prepared by microfluidization of centrifuged mixtures can be selected as the best conditions for encapsulation of cherry laurel powder by considering their total antioxidant activity. In this study, it was shown that capsules prepared by these methods can be recommended to be incorporated into foods in order to enhance their functionality by increasing antioxidant activity.

Keywords: antioxidant activity, cherry laurel, microencapsulation, microfluidization

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Authors: A. Pornpitakdumrong

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The aim of this research study was to develop recipe of Karanda ice cream as healthy promoting ice cream by high protein, low fat and naturally raw material, which found in local area. The results were found that appropriate condition for Karanda ice cream including incubation period, temperature and frozen time, which were 8-12 hours, -20 to -25 °C and 2-4 hours, respectively. Small fruit variety Karanda should selected only ripe fruits for Karanda ice cream made. Because of unripe fruits were contained resin and need to be air dried for reducing level of resin. Therefore, large fruit variety Karanda can be use both ripe and unripe fruits for Karanda ice cream made by without any astringent and bitter taste. However, small fruit variety Karanda was proper to made ice cream for trade, because occurring of industry to select the ripe fruits and commercially frozen, which be providing for the whole year compared with large variety fruits were rarely, low harvesting amount and short shelf life. Karanda ice cream produced from flesh part was attractive but was not accepted by consumers. It may due to resin contained with Karanda pulp, which led to be rough texture of ice cream. We were choose only Karanda juice, which was more appropriated and used Karanda juice with water by 1:1 ratio, because undiluted juice was sour taste. Most acceptance recipe of karanda ice cream product was sixth recipe by 91% of consumers, which was contained soy protein to made ice cream was delicate and swell, milk powder (little amount) to made ice cream was greasy, corn powder as stabilizer and undiluted coconut milk (little amount) to improve ice cream odor and similar to apricot odor.

Keywords: karonda fruits, Carissa carandas Linn, ice cream, healthy ice cream

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Authors: Hazrat Salman Sidique, Muhammad Tahir Khan, Haq Aman Ullah, Muhammad Mobashar, Muhammad Ishtiaq Sohail Mehmood

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The poor quality conventional feed for the livestock production in Pakistan are wheat straw, tops of sugar cane and tree leaves. To enhance the nutritive value of feed, this study focused on investigating the effects of fibrolytic enzyme (Fibrozyme®, Alltech Inc. Company, USA) at different levels i.e. 0, 5, 10, and 15g/kg of total mix ration on feed intake, digestibility, milk yield and composition, and economics of the ration in Holstein Friesians cows. Twelve Holstein Friesians cows of almost the same age, and lactation stage were randomly allocated into 4 equal groups i.e. A, B, C, and D. Four experimental rations supplemented with Fibrozyme® 0g, 5g, 10g, and 15g/Kg of total mix ration were assigned to these sets correspondingly. The dry matter intake was linearly and significantly (P<0.05) improved. A significant effect of Fibrozyme® was observed for organic matter digestibility, ether extract digestibility, crude fiber digestibility, nitrogen free extract digestibility, and acid detergent fiber digestibility while the results were statistically non-significant for crude protein digestibility, neutral detergent fiber digestibility, and ash digestibility. Milk yield and composition except fat were significantly (P<0.05) increased in all Fibrozyme® treated groups. This study concludes that supplementation of Fibrozyme® at the rate of 15g/Kg total mix ration improved the dry matter intake, nutrients digestibility, and milk production and constituents like protein, lactose, and solid not fat. Therefore, treatment of total mix ration with Fibrozyme® was desirably reasonable and profitable.

Keywords: digestibility, fibrozyme, TMR, digestibility, lactating cow

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Authors: Yujie Yuan, Yiyang Fan, Hong Fan

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Objective: The RNA methylation recognition protein Aly/REF export factor (ALYREF) is considered one type of “reader” protein acting as a recognition protein of m5C, has been reported involved in several biological progresses including cancer initiation and progression. 5-methylcytosine (m5C) is a conserved and prevalent RNA modification in all species, as accumulating evidence suggests its role in the promotion of tumorigenesis. It has been claimed that ALYREF mediates nuclear export of mRNA with m5C modification and regulates biological effects of cancer cells. However, the systematical regulatory pathways of ALYREF in cancer tissues have not been clarified, yet. Methods: The expression level of ALYREF in pan-cancer and their normal tissues was compared through the data acquired from The Cancer Genome Atlas (TCGA). The University of Alabama at Birmingham Cancer data analysis Portal UALCAN was used to analyze the relationship between ALYREF and clinical pathological features. The relationship between the expression level of ALYREF and prognosis of pan-cancer, and the correlation genes of ALYREF were figured out by using Gene Expression Correlation Analysis database GEPIA. Immune related genes were obtained from TISIDB (an integrated repository portal for tumor-immune system interactions). Immune-related research was conducted by using Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) and TIMER. Results: Based on the data acquired from TCGA, ALYREF has an obviously higher-level expression in various types of cancers compared with relevant normal tissues excluding thyroid carcinoma and kidney chromophobe. The immunohistochemical images on The Human Protein Atlas showed that ALYREF can be detected in cytoplasm, membrane, but mainly located in nuclear. In addition, a higher expression level of ALYREF in tumor tissue generates a poor prognosis in majority of cancers. According to the above results, cancers with a higher expression level of ALYREF compared with normal tissues and a significant correlation between ALYREF and prognosis were selected for further analysis. By using TISIDB, we found that portion of ALYREF co-expression genes (such as BIRC5, H2AFZ, CCDC137, TK1, and PPM1G) with high Pearson correlation coefficient (PCC) were involved in anti-tumor immunity or affect resistance or sensitivity to T cell-mediated killing. Furthermore, based on the results acquired from GEPIA, there was significant correlation between ALYREF and PD-L1. It was exposed that there is a negative correlation between the expression level of ALYREF and ESTIMATE score. Conclusion: The present study indicated that ALYREF plays a vital and universal role in cancer initiation and progression of pan-cancer through regulating mitotic progression, DNA synthesis and metabolic process, and RNA processing. The correlation between ALYREF and PD-L1 implied ALYREF may affect the therapeutic effect of immunotherapy of tumor. More evidence revealed that ALYREF may play an important role in tumor immunomodulation. The correlation between ALYREF and immune cell infiltration level indicated that ALYREF can be a potential therapeutic target. Exploring the regulatory mechanism of ALYREF in tumor tissues may expose the reason for poor efficacy of immunotherapy and offer more directions of tumor treatment.

Keywords: ALYREF, pan-cancer, immunotherapy, PD-L1

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Authors: Yamina Boukari, Nashiru Billa, Andrew Morris, Stephen Doughty, Kevin Shakesheff

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Poly (lactic-co-glycolic) acid (PLGA) is a synthetic polymer that can be used in bone tissue engineering with the aim of creating a scaffold in order to support the growth of cells. The formation of microspheres from this polymer is an attractive strategy that would allow for the development of an injectable system, hence avoiding invasive surgical procedures. The aim of this study was to develop a microsphere delivery system for use as an injectable scaffold in bone tissue engineering and evaluate various formulation parameters on its properties. Porous and lysozyme-containing PLGA microspheres were prepared using the double emulsion solvent evaporation method from various molecular weights (MW). Scaffolds were formed by sintering to contain 1 -3mg of lysozyme per gram of scaffold. The mechanical and physical properties of the scaffolds were assessed along with the release of lysozyme, which was used as a model protein. The MW of PLGA was found to have an influence on microsphere size during fabrication, with increased MW leading to an increased microsphere diameter. An inversely proportional relationship was displayed between PLGA MW and mechanical strength of formed scaffolds across loadings for low, intermediate and high MW respectively. Lysozyme release from both microspheres and formed scaffolds showed an initial burst release phase, with both microspheres and scaffolds fabricated using high MW PLGA showing the lowest protein release. Following the initial burst phase, the profiles for each MW followed a similar slow release over 30 days. Overall, the results of this study demonstrate that lysozyme can be successfully incorporated into porous PLGA scaffolds and released over 30 days in vitro, and that varying the MW of the PLGA can be used as a method of altering the physical properties of the resulting scaffolds.

Keywords: bone, microspheres, PLGA, tissue engineering

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Authors: Faheema Khan

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To investigate the ability of sensitive and tolerant genotype of Glycine max to adapt to a saline environment in a field, we examined the growth performance, water relation and activities of antioxidant enzymes in relation to photosynthetic rate, chlorophyll a fluorescence, photosynthetic pigment concentration, protein and proline in plants exposed to salt stress. Ten soybean genotypes (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712) were selected and grown hydroponically. After 3 days of proper germination, the seedlings were transferred to Hoagland’s solution (Hoagland and Arnon 1950). The growth chamber was maintained at a photosynthetic photon flux density of 430 μmol m−2 s−1, 14 h of light, 10 h of dark and a relative humidity of 60%. The nutrient solution was bubbled with sterile air and changed on alternate days. Ten-day-old seedlings were given seven levels of salt in the form of NaCl viz., T1 = 0 mM NaCl, T2=25 mM NaCl, T3=50 mM NaCl, T4=75 mM NaCl, T5=100 mM NaCl, T6=125 mM NaCl, T7=150 mM NaCl. The investigation showed that genotype Pusa-24, PK-416 and Pusa-20 appeared to be the most salt-sensitive. genotypes as inferred from their significantly reduced length, fresh weight and dry weight in response to the NaCl exposure. Pusa-37 appeared to be the most tolerant genotype since no significant effect of NaCl treatment on growth was found. We observed a greater decline in the photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, in salt-sensitive (Pusa-24) genotype than in salt-tolerant Pusa-37 under high salinity. Numerous primers were verified on ten soybean genotypes obtained from Operon technologies among which 30 RAPD primers shown high polymorphism and genetic variation. The Jaccard’s similarity coefficient values for each pairwise comparison between cultivars were calculated and similarity coefficient matrix was constructed. The closer varieties in the cluster behaved similar in their response to salinity tolerance. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings.Salt tolerant genotype Pusa-37, was further analysed by 2-Dimensional gel electrophoresis to analyse the differential expression of proteins at high salt stress. In the Present study, 173 protein spots were identified. Of these, 40 proteins responsive to salinity were either up- or down-regulated in Pusa-37. Proteomic analysis in salt-tolerant genotype (Pusa-37) led to the detection of proteins involved in a variety of biological processes, such as protein synthesis (12 %), redox regulation (19 %), primary and secondary metabolism (25 %), or disease- and defence-related processes (32 %). In conclusion, the soybean plants in our study responded to salt stress by changing their protein expression pattern. The photosynthetic, biochemical and molecular study showed that there is variability in salt tolerance behaviour in soybean genotypes. Pusa-24 is the salt-sensitive and Pusa-37 is the salt-tolerant genotype. Moreover this study gives new insights into the salt-stress response in soybean and demonstrates the power of genomic and proteomic approach in plant biology studies which finally could help us in identifying the possible regulatory switches (gene/s) controlling the salt tolerant genotype of the crop plants and their possible role in defence mechanism.

Keywords: glycine max, salt stress, RAPD, genomic and proteomic variability

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Authors: Galuh Asri Bestari, Hajar Shofiyya

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Double burden of malnutrition (DBM) has been a global problem in these last decades occurs in both developed and developing countries. Overweight in adults and stunting among preschool children have dramatically increased and become the main problems of malnutrition that should be solved immediately since they are directly related with the health status and productivity. Reformulation of food product by using the local sea resources called terasi shrimp (Acetes indicus) has a potential possibility in facing the DBM. A study was carried out in Indonesia to determine the acceptability of terasi shrimp powder through sensory evaluation. Terasi shrimps were processed into powder form through sun drying and pounding methods. The powder form was directly added in food as alternative seasonings and tested among stunted and normal preschool children. Meanwhile, a further processing method is given to the shrimp powder tested in overweight and normal-weighed adults. The shrimp powder was mixed with sago flour and formed into balls, then steamed for 15-20 minutes, and finally served as alternative snacks. Based on the sensory evaluation, the shrimp powder has a good acceptance in taste (54%), shape (60%), and color properties (63%), while the shrimp balls has a good acceptance in size (65%), shape (50%), color (48%), taste (40%), and texture (36%). Terasi shrimp powder can be stored for a month in room temperature. In addition, carried out chemical analysis revealed that terasi shrimp (Acetes indicus) has higher percentage of protein, calcium, and iron than other animal sources, but conversely contains zero sodium and very low percentage of fat. Terasi shrimp’s shell also contains a substance called chitosan which acts by forming gels in the intestinal tract to entrap lipids, thus interfering with their absorption. After going through some processing methods, the shrimp powder and balls did not show any significant changes in their nutrient contents. So that, terasi shrimp powder is good to be consumed not only by overweight adults, but also by children to support their optimum growth. Intervention of terasi shrimp powder should be implemented step by step from national up to global governance program to face the DBM.

Keywords: Acetes indicus, alternative food, double burden of malnutrition, sensory evaluation

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Authors: Lawrence Gumbe

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Kenya Vision 2030 is the country's programme for transformation covering the period 2008 to 2030. Its objective is to help transform Kenya into a newly industrializing, middle-income, exceeding US$10000, country providing a high quality of life to all its citizens by 2030, in a clean and secure environment. Increased agricultural and production and productivity is crucial for the realization of Vision 2030. Mechanization of agriculture in order to achieve greater yields is the only way to achieve these objectives. There are contending groups and views on the strategy for agricultural mechanization. The first group are those who oppose the widespread adoption of advanced technologies (mostly internal combustion engines and tractors) in agricultural mechanization as entirely inappropriate in most situations in developing countries. This group argues that mechanically powered -agricultural mechanization often leads to displacement of labour and hence increased unemployment, and this results in a host of other socio-economic problems, amongst them, rural-urban migration, inequitable distribution of wealth and in many cases an increase in absolute poverty, balance of payments due to the need to import machinery, fuel and sometimes technical assistance to manage them. The second group comprises of those who view the use of the improved hand tools and animal powered technology as transitional step between the most rudimentary step in technological development (characterized by entire reliance on human muscle power) and the advanced technologies (characterized 'by reliance on tractors and other machinery). The third group comprises those who regard these intermediate technologies (ie. improved hand tools and draught animal technology in agriculture) as a ‘delaying’ tactic and they advocate the use of mechanical technologies as-the most appropriate. This group argues that alternatives to the mechanical technologies do not just exist as a practical matter, or, if they are available, they are inefficient and they cannot be compared to the mechanical technologies in terms of economics and productivity. The fourth group advocates a compromise between groups two and third above. This group views the improved hand tools and draught animal technology as more of an 18th century technology and the modem tractor and combine harvester as too advanced for developing countries. This group has been busy designing an ‘intermediate’, ‘appropriate’, ‘mini’, ‘micro’ tractor for use by farmers in developing countries. This paper analyses and concludes on the different agricultural mechanization strategies available to Kenya and other third world countries

Keywords: agriculture, mechanazation, transformation, industrialization

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Authors: Kun Hua O., Dong Woon Kim, Won Hyung Lee

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Introduction: Neuropathic pain induced by spinal or peripheral nerve injury is highly resistant to common painkillers, nerve blocks, and other pain management approaches. Recently, several new therapeutic drug candidates have been developed to control neuropathic pain. In this study, we used the spinal nerve L5 ligation (SNL) model to investigate the ability of intrathecal or intravenous Evans blue to decrease pain behavior and to study the relationship between Evans blue and the neural structure of pain transmission. Method: Neuropathic pain (allodynia) of the left hind paw was induced by unilateral SNL in Sprague-Dawley rats(n=10) in each group. Evans blue (5, 15, 50μg/10μl) or phosphate buffer saline(PBS,10μl) was injected intrathecally at 3days post-ligation or intravenously(1mg/200 μl) 3days and 5days post-ligation . Mechanical sensitivity was assessed using Von Frey filaments at 3 days post-ligation and at 2 hours, days 1, 2, 3, 5,7 after intrathecal Evans blue injection, and on days 2, 4, 7, and 11 at 14 days after intravenous injection. In the intrathecal group, microglia and glutaminergic neurons in the dorsal horn and VNUT(vesicular nucleotide transporter) in the dorsal root ganglia were tested to evaluate co-staining with Evans blue. The experimental procedures were performed in accordance with the animal care guideline of the Korean Academy of Medical Science(Animal ethic committee of Chungnam National University Hospital: CNUH-014-A0005-1). Results: Tight ligation of the L5 spinal nerve induced allodynia in the left hind paw 3 days post-ligation. Intrathecal Evans blue most significantly(P＜0.001) alleviated allodynia at 2 days after intrathecal, but not an intravenous injection. Glutaminergic neurons in the dorsal horn and VNUT in the dorsal root ganglia were co-stained with Evans blue. On the other hand, microglia in the dorsal horn were partially co-stained with Evans blue. Conclusion: We confirmed that Evans blue might have an analgesic effect through the central nervous system, not another system in neuropathic pain of the SNL animal model. These results suggest Evans blue may be a potential new drug for the treatment of chronic pain. This research was supported by the National Research Foundation of Korea (NRF-2020R1A2C100757512), funded by the Ministry of Education.

Keywords: neuropathic pain, Evas blue, intrathecal, intravenous

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Authors: Javier Enciso-Benavides, Fredy Fabian, Carlos Castaneda, Luis Alfaro, Alex Choque, Aparicio Aguilar, Javier Enciso

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Background: The use of biomarkers in breast cancer diagnosis, therapy, and prognosis has gained increasing interest. Cancer stem cells (CSCs) are a subpopulation of tumor cells that can drive tumor initiation and may cause relapse. Therefore, due to the importance of diagnosis, therapy, and prognosis, several biomarkers that characterize CSCs have been identified; however, in treatment-naïve triple-negative breast tumors, there is an urgent need to identify new biomarkers and therapeutic targets. According to this, the aim of this study was to identify serum proteins associated with cancer stem cells and pluripotency in women with triple-negative breast tumors in order to subsequently identify a biomarker for this type of breast tumor. Material and Methods: Whole blood samples from 12 women with histopathologically diagnosed triple-negative breast tumors were used after obtaining informed consent from the patient. Blood serum was obtained by conventional procedure and frozen at -80ºC. Identification of cancer stem cell-associated proteins was performed by matrix-assisted laser desorption/ionisation-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF MS), protein analysis was obtained using the AB Sciex TOF/TOF™ 5800 system (AB Sciex, USA). Sequences not aligned by ProteinPilot™ software were analyzed by Protein BLAST. Results: The following proteins related to pluripotency and cancer stem cells were identified by MALDI TOF/TOF mass spectrometry: A-chain, Serpin A12 [Homo sapiens], AIEBP [Homo sapiens], Alpha-one antitrypsin, AT {internal fragment} [human, partial peptide, 20 aa] [Homo sapiens], collagen alpha 1 chain precursor variant [Homo sapiens], retinoblastoma-associated protein variant [Homo sapiens], insulin receptor, CRA_c isoform [Homo sapiens], Hydroxyisourate hydrolase [Streptomyces scopuliridis], MUCIN-6 [Macaca mulatta], Alpha-actinin-3 [Chrysochloris asiatica], Polyprotein M, CRA_d isoform, partial [Homo sapiens], Transcription factor SOX-12 [Homo sapiens]. Recommendations: The serum proteins identified in this study should be investigated in the exosome of triple-negative breast cancer stem cells and in the blood serum of women without breast cancer. Subsequently, proteins found only in the blood serum of women with triple-negative breast cancer should be identified in situ in triple-negative breast cancer tissue in order to identify a biomarker to study the evolution of this type of cancer, or that could be a therapeutic target. Conclusions: Eleven cancer stem cell-related serum proteins were identified in 12 women with triple-negative breast cancer, of which MUCIN-6, retinoblastoma-associated protein variant, transcription factor SOX-12, and collagen alpha 1 chain are the most representative and have not been studied so far in this type of breast tumor. Acknowledgement: This work was supported by Proyecto CONCYTEC–Banco Mundial “Mejoramiento y Ampliacion de los Servicios del Sistema Nacional de Ciencia Tecnología e Innovacion Tecnologica” 8682-PE (104-2018-FONDECYT-BM-IADT-AV).

Keywords: triple-negative breast cancer, MALDI TOF/TOF MS, serum proteins, cancer stem cells

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Authors: Ilzira A. Minigalieva, Tatiana V. Bushueva, Eleonore Frohlich, Vladimir Panov, Ekaterina Shishkina, Boris A. Katsnelson

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Background: The overwhelming majority of the experimental studies in the field of metal nanotoxicology have been performed on cultures of established cell lines, with very few researchers focusing on animal experiments, while a juxtaposition of conclusions inferred from these two types of research is blatantly lacking. The least studied aspect of this problem relates to characterizing and predicting the combined toxicity of metallic nanoparticles. Methods: Comparative and combined toxic effects of purposefully prepared spherical NiO and Mn₃O₄ nanoparticles (mean diameters 16.7 ± 8.2 nm and 18.4 ± 5.4 nm respectively) were estimated on cultures of human cell lines: MRC-5 fibroblasts, THP-1 monocytes, SY-SY5Y neuroblastoma cells, as well as on the latter two lines differentiated to macrophages and neurons, respectively. The combined cytotoxicity was mathematically modeled using the response surface methodology. Results: The comparative assessment of the studied NPs unspecific toxicity previously obtained in vivo was satisfactorily reproduced by the present in vitro tests. However, with respect to manganese-specific brain damage which had been demonstrated by us in animal experiment with the same NPs, the testing on neuronall cell culture showed only a certain enhancing effect of Mn₃O₄-NPs on the toxic action of NiO-NPs, while the role of the latter prevailed. Conclusion: From the point of view of the preventive toxicology, the experimental modeling of metallic NPs combined toxicity on cell cultures can give non-reliable predictions of the in vivo action’s effects.

Keywords: manganese oxide, nickel oxide, nanoparticles, in vitro toxicity

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Authors: Bhawna Chandravanshi, Ramesh Bhonde

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In the present study, we focused on the improvisation of islet survival in hypoxia. The Islet-like cell aggregates (ICAs) derived from Wharton's jelly mesenchymal stem cells (WJ-MSC) were cultured with and without WJ-MSC for 48h in hypoxia and normoxia and tested for their direct trophic effect on β cell survival. The WJ MSCs themselves secreted insulin upon glucose challenge and expressed the pancreatic markers at both transcription and translational level (C-peptide, Insulin, Glucagon and Glut 2). Direct contact of MSCs with ICAs facilitate the highest viability under hypoxia as evidenced by fluorescein diacetate/propidium iodide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cytokine analysis of the co-cultured ICAs revealed amplification of anti-inflammatory cytokine-like TGFβ and TNFα accompanied by depletion of pro-inflammatory cytokines. The increment in VEGF and PDGFa was also seen showing their ability to vascularize upon transplantation. This was further accompanied by reduction in total reactive oxygen species, nitric oxide, and super oxide ions and down-regulation of Caspase3, Caspase8, p53 and up regulation of Bcl2 confirming prevention of apoptosis in ICAs. There was a significant reduction in the expression of p38 protein in the presence of MSCs making the ICAs responsive to glucose. Taken together our data demonstrate for the first time that the WJ-MSC expressed pancreatic markers and their supplementation protected engineered islets against hypoxia, oxidative stress, and inflammatory cytokines by inhibiting p38 MAPK protein.

Keywords: hypoxia, islet-like cell aggregates, inflammatory cytokines, oxidative stress

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Authors: Laurentius J. M. Rumokoy, Jimmy Posangi, Wisje Lusia Toar, Julio Lopez Aban

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The threat of pathogen infection agents to the neonates is a major health problem to the new born life livestock. Neonate losses became an important case in the world as well as in Indonesia. This condition can be triggered by an infection with nematode in conjunction with a failure of immunoglobulin passive transfer. The study was conducted to evaluate the role of the curcuma combined with IgG colostrum on the development of parasites in the gut of CD1 mice. Animal experiments were divided in four groups (G) based on the treatment: G1 (infection only); G2 (curcuma+infection), G3 (IgG + infection) and G4 (curcuma+IgG+infection). The parameters measured were EPG (eggs per gram) and female in the intestine. The results obtained showed that the treatment has no a significant influence on the number of eggs per gram of feces in the group infected compared to the control group without receiving IgG nor curcuma. However, the EGP response tended to decrease at day 6 in G3 and G4 with a minimum number at zero eggs. This performant showed that the immunoglobulin-G and curcuma substances could slightly decreased the number of eggs in animal infected with Strongyloides. The results obtained showed also that the treatment has no significant difference (P > 0.05) on female larva in the gut of MCD1 experimental. In other side, we found that the best performance to inhibit the female quantity in the gut was the treatment with IgG and infection of parasite in G3. In this treatment, the minimum number was five female only in the gut. The results described IgG response was better than the curcuma single use in reducing the female parasite in the gut. This positive response of IgG compared to other controls group was associated with the function of colostrum antibodies.

Keywords: parasites, livestock, curcuma, colostrums

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Authors: M. Cabral, A. Verdin, G. Garçon, A. Touré, C. Diop, M. Fall, S. Bouhsina, D. Dewaele, F.Cazier, A. Tall Dia, P. Shirali, A. Diouf

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This case-control study dealt with the health adverse effects within the population neighboring the Mbeubeuss waste dump, which is located near the district of Malika (Diamalaye II) in Dakar (Senegal). All the household and industrial waste arising from Dakar are stored in this open landfill without being covered and are therefore possible sources of Pb and Cd contaminated air emissions and lixiviates. The objective of this study is part of improving the health of the population neighboring Mbeubeuss by determining Pb and Cd concentrations both in environment and humans, and studying possible renal function alterations within the adults. Soil and air samples were collected in the control site (Darou Salam) and the waste dump neighboring site (Diamalaye II). Control and exposed adults were recruited as living in Darou Salam (n = 52) and in Diamalaye II (n = 77). Pb and Cd concentrations in soil, air and biological samples were determined. Moreover, we were interested in analyzing some impregnation (zinc protoporphyrin, d-aminolevulinic acid dehydratase) and oxidative stress biomarkers (malonedialdehyde, gluthatione status), in addition to several nephrotoxicity parameters (creatinuria, proteinuria, lactate dehydrogenase, CC16 protein, glutathione S-transferase-alpha and retinol binding protein) in blood and/or urine. The results showed the significant Pb and Cd contamination of the soil and air samples derived from the landfill, and therefore of the neighboring population of adults. This critical exposure to environmental Pb and Cd had some harmful consequences for their health, as shown by the reported oxidative stress and nephrotoxicity signs.

Keywords: Pb and Cd environmental exposure, impregnation markers, landfill, nephrotoxicity markers

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Authors: Yew Mun Yip, Dawei Zhang

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Proteins are the biological machinery that executes specific vital functions in every cell of the human body by folding into their 3D structures. When a protein misfolds from its native structure, the machinery will malfunction and lead to misfolding diseases. Although in vitro experiments are able to conclude that the mutations of the amino acid sequence lead to incorrectly folded protein structures, these experiments are unable to decipher the folding process. Therefore, molecular dynamic (MD) simulations are employed to simulate the folding process so that our improved understanding of the folding process will enable us to contemplate better treatments for misfolding diseases. MD simulations make use of force fields to simulate the folding process of peptides. Secondary structures are formed via the hydrogen bonds formed between the backbone atoms (C, O, N, H). It is important that the hydrogen bond energy computed during the MD simulation is accurate in order to direct the folding process to the native structure. Since the atoms involved in a hydrogen bond possess very dissimilar electronegativities, the more electronegative atom will attract greater electron density from the less electronegative atom towards itself. This is known as the polarization effect. Since the polarization effect changes the electron density of the two atoms in close proximity, the atomic charges of the two atoms should also vary based on the strength of the polarization effect. However, the fixed atomic charge scheme in force fields does not account for the polarization effect. In this study, we introduce the polarized structure-specific backbone charge (PSBC) model. The PSBC model accounts for the polarization effect in MD simulation by updating the atomic charges of the backbone hydrogen bond atoms according to equations derived between the amount of charge transferred to the atom and the length of the hydrogen bond, which are calculated from quantum-mechanical calculations. Compared to other polarizable models, the PSBC model does not require quantum-mechanical calculations of the peptide simulated at every time-step of the simulation and maintains the dynamic update of atomic charges, thereby reducing the computational cost and time while accounting for the polarization effect dynamically at the same time. The PSBC model is applied to two different β-peptides, namely the Beta3s/GS peptide, a de novo designed three-stranded β-sheet whose structure is folded in vitro and studied by NMR, and the trpzip peptides, a double-stranded β-sheet where a correlation is found between the type of amino acids that constitute the β-turn and the β-propensity.

Keywords: hydrogen bond, polarization effect, protein folding, PSBC

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Authors: Nicole Selvi Hill, Archchana Radhakrishnan

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Introduction: Traumatic brain injury (TBI) is a leading cause of mortality among trauma patients. In the management of TBI, the main principle is avoiding cerebral ischemia, as this is a strong determiner of neurological outcomes. The use of vasoactive drugs, such as vasopressin, has an important role in maintaining cerebral perfusion pressure to prevent secondary brain injury. Current guidelines do not suggest a preferred vasoactive drug to administer in the management of TBI, and there is a paucity of information on the therapeutic potential of vasopressin following TBI. Vasopressin is also an endogenous anti-diuretic hormone (AVP), and pathways mediated by AVP play a large role in the underlying pathological processes of TBI. This creates an overlap of discussion regarding the therapeutic potential of vasopressin following TBI. Currently, its popularity lies in vasodilatory and cardiogenic shock in the intensive care setting, with increasing support for its use in haemorrhagic and septic shock. Methodology: This is a review article based on a literature review. An electronic search was conducted via PubMed, Cochrane, EMBASE, and Google Scholar. The aim was to identify clinical studies looking at the therapeutic administration of vasopressin in severe traumatic brain injury. The primary aim was to look at the neurological outcome of patients. The secondary aim was to look at surrogate markers of cerebral perfusion measurements, such as cerebral perfusion pressure, cerebral oxygenation, and cerebral blood flow. Results: Eight papers were included in the final number. Three were animal studies; five were human studies, comprised of three case reports, one retrospective review of data, and one randomised control trial. All animal studies demonstrated the benefits of vasopressors in TBI management. One animal study showed the superiority of vasopressin in reducing intracranial pressure and increasing cerebral oxygenation over a catecholaminergic vasopressor, phenylephrine. All three human case reports were supportive of vasopressin as a rescue therapy in catecholaminergic-resistant hypotension. The retrospective review found vasopressin did not increase cerebral oedema in TBI patients compared to catecholaminergic vasopressors; and demonstrated a significant reduction in the requirements of hyperosmolar therapy in patients that received vasopressin. The randomised control trial results showed no significant differences in primary and secondary outcomes between TBI patients receiving vasopressin versus those receiving catecholaminergic vasopressors. Apart from the randomised control trial, the studies included are of low-level evidence. Conclusion: Studies favour vasopressin within certain parameters of cerebral function compared to control groups. However, the neurological outcomes of patient groups are not known, and animal study results are difficult to extrapolate to humans. It cannot be said with certainty whether vasopressin’s benefits stand above usage of other vasoactive drugs due to the weaknesses of the evidence. Further randomised control trials, which are larger, standardised, and rigorous, are required to improve knowledge in this field.

Keywords: catecholamines, cerebral perfusion pressure, traumatic brain injury, vasopressin, vasopressors

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Authors: Hassan M. H. Mustafa

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This piece of research addresses an interesting comparative analytical study. Which considers two concepts of diverse algorithmic computational intelligence approaches related tightly with Neural and Non-Neural Systems. The first algorithmic intelligent approach concerned with observed obtained practical results after three neural animal systems’ activities. Namely, they are Pavlov’s, and Thorndike’s experimental work. Besides a mouse’s trial during its movement inside figure of eight (8) maze, to reach an optimal solution for reconstruction problem. Conversely, second algorithmic intelligent approach originated from observed activities’ results for Non-Neural Ant Colony System (ACS). These results obtained after reaching an optimal solution while solving Traveling Sales-man Problem (TSP). Interestingly, the effect of increasing number of agents (either neurons or ants) on learning performance shown to be similar for both introduced systems. Finally, performance of both intelligent learning paradigms shown to be in agreement with learning convergence process searching for least mean square error LMS algorithm. While its application for training some Artificial Neural Network (ANN) models. Accordingly, adopted ANN modeling is a relevant and realistic tool to investigate observations and analyze performance for both selected computational intelligence (biological behavioral learning) systems.

Keywords: artificial neural network modeling, animal learning, ant colony system, traveling salesman problem, computational biology

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Authors: Ubani Onyedikachi, Atagana Harrison Ifeanyichukwu, Thantsha Mapitsi Silvester

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Components of oil sludge (PAHs) are known cytotoxic, mutagenic and potentially carcinogenic compounds also bacteria and fungi have been found to degrade PAHs to innocuous compounds. This study is aimed at measuring the effect of pig, cow, horse and poultry manures on the degradation in selected PAHs present in oil sludge. Soil spiked with oil sludge was co-composted differently with each manure in a ratio of 2:1 (w/w) spiked soil: manure and wood-chips in a ratio of 2:1 (w/v) spiked soil: wood-chips. Control was set up similar as the one above but without manure. The mixtures were incubated for 10 months at room temperature. Compost piles were turned weekly and moisture level was maintained at between 50% and 70%. Moisture level, pH, temperature, CO2 evolution and oxygen consumption were measured monthly and the ash content at the end of experimentation. Highest temperature reached was 27.5 °C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2 evolution was highest in poultry manure at 18.78μg/dwt/day. Microbial growth and activities were enhanced; bacteria identified were Bacillus, Arthrobacter and Staphylococcus species. Percentage reduction in PAHs was measured using automated soxhlet extractor with Dichloromethane coupled with gas chromatography/mass spectrometry (GC/MS). Results from PAH measurements showed reduction between 77% and 99%. Co-composting of spiked soils with animal manures enhanced the reduction in PAHs.

Keywords: animal manures, bioremediation, co-composting, oil refinery sludge, PAHs

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Authors: Mahzad Yousefi, Azin Tavakoli

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Periodontal disease is an inflammatory reaction; therefore, it is predicted that changes may occur in some inflammatory parameters that can be detected in routine blood tests. The objective of this study was to evaluate the hematological and biochemistry changes that occur in dogs affected with advanced stages of periodontal disease. 87 dogs were diagnosed with periodontal disease (PD group), and 76 healthy dogs entered the study. The PD dogs had been affected with periodontitis stage 3 or 4 and were candidates for any dental extractions. The healthy dogs were either referred for annual checkups or for issuing health travel certificates that their owners asked for complete lab tests. Neither the diseased nor healthy subjects had a history of infectious, or other general health problems or surgery in the past 3 months. Age, as well as all hematologic including PCV, WBC and RBC count, Hb, MCV, MCH, MCHC, PLT, CBC, NLR, and biochemistry data, including total protein, albumin, glucose, BUN, Creatinine, ALT, AST, and ALP, were recorded and analyzed statistically. Results confirmed that aging has a significant direct relationship with the advanced stages of periodontal disease. Mild leukocytosis occurred in the diseased group; however, it was not significant. Also, the mean total protein of the PD group was lower than that of the healthy dogs, and serum levels of albumin were found to be lower significantly in the diseased group (P<0.05). Mean ±SD amount of Platelet, MCH, and ALT were significantly higher in the diseased group in comparison to the healthy dogs (P<0.05). No significant differences were reported in other evaluated parameters. It is concluded that CBC indices of PD dogs are not systemic inflammatory; however, only a decrease in albumin showed inflammatory responses. Some indices in routine laboratory tests can be changed significantly during advanced stages of the periodontal disease dogs.

Keywords: periodontal disease, dogs, hematological factors, advanced stages, blood tests

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Authors: Aarushi Agarwal, Tara Singh, I.L Singh, Anju Lata Singh, Trayambak Tiwari

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Simple discrimination in near absence of attention has been widely observed. Dual-task studies with natural scenes studies have been claimed as being preattentive in nature that facilitated categorization simultaneously with the attentional demanding task. So in this study, multiple images at the periphery are presented, initiating parallel processing in near absence of attention. For the central demanding task rotated letters were presented in both conditions, while in periphery natural and animal images were presented. To understand the breakpoint of ability to perform in near absence of attention one, two and three peripheral images were presented simultaneously with central task and subjects had to respond when all belong to the same category. Individual participant performance did not show a significant difference in both conditions central and peripheral task when the single peripheral image was shown. In case of two images high-level parallel processing could take place with little attentional resources. The eye tracking results supports the evidence as no major saccade was made in a large number of trials. Three image presentations proved to be a breaking point of the capacities to perform outside attentional assistance as participants showed a confused eye gaze pattern which failed to make the natural and animal image discriminations. Thus, we can conclude attention and awareness being independent mechanisms having limited capacities.

Keywords: attention, dual task pardigm, parallel processing, break point, saccade

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Authors: M. Y. Abubakar, M. Yunisa, A. N. Muhammad

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Constraining the production Clarias gariepinus and Oreochromis niloticus culture is the prohibitive cost of feed. We assess the performance of the species fingerlings on diets substituted with composite. Four dietary treatments (0%, 25%, 45%, and 75%) for C. gariepinus and five (0%, 25%, 50%, 75%, and whole food composite) for O. niloticus were formulated and each fed to 15 fingerlings for C. gariepinus and 10 fingerlings for O. niloticus stocked in 75ltrs plastic bowls, replicated trice in a completely randomized design. The experiment lasted 56 days. Percent survival rate was significantly (p < 0.05) higher (57.78 ± 9.69) in C. gariepinus fed diet III. The growth and nutrient utilization indices were least in the fish fed diet IV, which was significantly (p < 0.05) lower than in other treatments. Fish fed dietary treatment III, recorded the best in growth and nutrient utilization indices and was significantly higher (p < 0.05) than those fed dietary treatments I & II which were non-significant (p > 0.05) and higher than those fed 75% substitution. Better profit index was in the fish fed diet with 50% substitution level. For O. niloticus, the survival (172.62 ± 39.03) was significantly higher (p < 0.05) in those fed 25% substituted diet. For growth indices, the least performed were those fed whole composite while other treatments were non-significant (p > 0.05) different from each other. In terms of nutrient utilization, fish fed diet substituted at 0%, 25%, 50% and 75% food composite had similar food conversion ratio and protein efficiency ratio. However, there was no significant difference in the profit index among the whole treatment. It can be concluded that food composite from Sokoto house-holds can optimally replace groundnut cake up to 50% level as a protein source in the diets of Clarias gariepinus and O. niloticus fingerlings without adverse effects on survival, growth, and nutrient utilization.

Keywords: food composite, nutrient utilization, C. gariepinus, O. niloticus household, substitution levels

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Authors: F. Yılmaz, Y. Saylan, A. Derazshamshir, S. Atay, A. Denizli

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Quartz crystal microbalance (QCM), high-resolution mass-sensing technique, measures changes in mass on oscillating quartz crystal surface by measuring changes in oscillation frequency of crystal in real time. Protein adsorption techniques via hydrophobic interaction between protein and solid support, called hydrophobic interaction chromatography (HIC), can be favorable in many cases. Some nanoparticles can be effectively applied for HIC. HIC takes advantage of the hydrophobicity of proteins by promoting its separation on the basis of hydrophobic interactions between immobilized hydrophobic ligands and nonpolar regions on the surface of the proteins. Lysozyme is found in a variety of vertebrate cells and secretions, such as spleen, milk, tears, and egg white. Its common applications are as a cell-disrupting agent for extraction of bacterial intracellular products, as an antibacterial agent in ophthalmologic preparations, as a food additive in milk products and as a drug for treatment of ulcers and infections. Lysozyme has also been used in cancer chemotherapy. The aim of this study is the synthesis of hydrophobic nanoparticles for Lysozyme detection. For this purpose, methacryoyl-L-phenylalanine was chosen as a hydrophobic matrix. The hydrophobic nanoparticles were synthesized by micro-emulsion polymerization method. Then, hydrophobic QCM nanosensor was characterized by Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, atomic force microscopy (AFM) and zeta size analysis. Hydrophobic QCM nanosensor was tested for real-time detection of Lysozyme from aqueous solution. The kinetic and affinity studies were determined by using Lysozyme solutions with different concentrations. The responses related to a mass (Δm) and frequency (Δf) shifts were used to evaluate adsorption properties.

Keywords: nanosensor, HIC, lysozyme, QCM

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Authors: Ved Vrat Verma, Rani Gupta, Manisha Goel

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γ-glutamyltranspeptidase (GGT: EC 2.3.2.2) is an N-terminal nucleophile hydrolase conserved in all three domains of life. GGT plays a key role in glutathione metabolism where it catalyzes the breakage of the γ-glutamyl bonds and transfer of γ-glutamyl group to water (hydrolytic activity) or amino acids or short peptides (transpeptidase activity). GGTs from bacteria, archaea, and eukaryotes (human, rat and mouse) are homologous proteins sharing >50% sequence similarity and conserved four layered αββα sandwich like three dimensional structural fold. These proteins though similar in their structure to each other, are quite diverse in their enzyme activity: some GGTs are better at hydrolysis reactions but poor in transpeptidase activity, whereas many others may show opposite behaviour. GGT is known to be involved in various diseases like asthma, parkinson, arthritis, and gastric cancer. Its inhibition prior to chemotherapy treatments has been shown to sensitize tumours to the treatment. Microbial GGT is known to be a virulence factor too, important for the colonization of bacteria in host. However, all known inhibitors (mimics of its native substrate, glutamate) are highly toxic because they interfere with other enzyme pathways. However, a few successful efforts have been reported previously in designing species specific inhibitors. We aim to leverage the diversity seen in GGT family (pathogen vs. eukaryotes) for designing specific inhibitors. Thus, in the present study, we have used DIVERGE software to identify sites in GGT proteins, which are crucial for the functional and structural divergence of these proteins. Since, type II divergence sites vary in clade specific manner, so type II divergent sites were our focus of interest throughout the study. Type II divergent sites were identified for pathogen vs. eukaryotes clusters and sites were marked on clade specific representative structures HpGGT (2QM6) and HmGGT (4ZCG) of pathogen and eukaryotes clade respectively. The crucial divergent sites within 15 A radii of the binding cavity were highlighted, and in-silico mutations were performed on these sites to delineate the role of these sites on the mechanism of catalysis and protein folding. Further, the amino acid network (AAN) analysis was also performed by Cytoscape to delineate assortative mixing for cavity divergent sites which could strengthen our hypothesis. Additionally, molecular dynamics simulations were performed for wild complexes and mutant complexes close to physiological conditions (pH 7.0, 0.1 M ionic strength and 1 atm pressure) and the role of putative divergence sites and structural integrities of the homologous proteins have been analysed. The dynamics data were scrutinized in terms of RMSD, RMSF, non-native H-bonds and salt bridges. The RMSD, RMSF fluctuations of proteins complexes are compared, and the changes at protein ligand binding sites were highlighted. The outcomes of our study highlighted some crucial divergent sites which could be used for novel inhibitors designing in a species-specific manner. Since, for drug development, it is challenging to design novel drug by targeting similar protein which exists in eukaryotes, so this study could set up an initial platform to overcome this challenge and help to deduce the more effective targets for novel drug discovery.

Keywords: γ-glutamyltranspeptidase, divergence, species-specific, drug design

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Authors: Francielo Vendruscolo

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During industrial processing of raw materials of animal and vegetable origin, large amounts of solid, liquid and gaseous wastes are generated. Solid residues are usually materials rich in carbohydrates, protein, fiber and minerals. Brewer’s spent grain (BSG) is the main waste generated in the brewing industry, representing 85% of the waste generated in this industry. It is estimated that world’s BSG generation is approximately 38.6 x 106 t per year and represents 20-30% (w/w) of the initial mass of added malt, resulting in low commercial value by-product, however, does not have economic value, but it must be removed from the brewery, as its spontaneous fermentation can attract insects and rodents. For every 100 grams in dry basis, BSG has approximately 68 g total fiber, being divided into 3.5 g of soluble fiber and 64.3 g of insoluble fiber (cellulose, hemicellulose and lignin). In addition to dietary fibers, depending on the efficiency of the grinding process and mashing, BSG may also have starch, reducing sugars, lipids, phenolics and antioxidants, emphasizing that its composition will depend on the barley variety and cultivation conditions, malting and technology involved in the production of beer. BSG demands space for storage, but studies have proposed alternatives such as the use of drying, extrusion, pressing with superheated steam, and grinding to facilitate storage. Other important characteristics that enhance its applicability in bioremediation, effluent treatment and biotechnology, is the surface area (SBET) of 1.748 m2 g-1, total pore volume of 0.0053 cm3 g-1 and mean pore diameter of 121.784 Å, characterized as a macroporous and possess fewer adsorption properties but have great ability to trap suspended solids for separation from liquid solutions. It has low economic value; however, it has enormous potential for technological applications that can improve or add value to this agro-industrial waste. Due to its composition, this material has been used in several industrial applications such as in the production of food ingredients, fiber enrichment by its addition in foods such as breads and cookies in bioremediation processes, substrate for microorganism and production of biomolecules, bioenergy generation, and civil construction, among others. Therefore, the use of this waste or by-product becomes essential and aimed at reducing the amount of organic waste in different industrial processes, especially in breweries.

Keywords: brewer’s spent malt, agro-industrial residue, lignocellulosic material, waste generation

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Authors: Shreya Ghosh, Akansha Garg, Chayanika Kala, Ashwani Kumar Thakur

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Background: Granuloma formation is one of the characteristic features of tuberculosis. Besides, chronic inflammation underlying tuberculosis is often indicated by an increase in the concentration of serum amyloid A (SAA) protein. The connection between tuberculosis and SAA-driven secondary amyloidosis is well documented. However, SAA-derived amyloid deposition start sites are not well understood in tuberculosis and other chronic inflammatory conditions. It was hypothesized that granuloma could be a potential site for an amyloid deposition because both SAA protein and proteases that cleave SAA into aggregation-prone fragments are reported to be present in the granuloma. Here the authors have shown the presence of SAA-derived amyloid deposits in the granuloma of tuberculosis patients. Methodology: Over a period of two years, tuberculosis patients were screened, and biopsies were collected from the affected organs of the patients. The gold standard, Congo red dye staining, was used to identify amyloid deposits in the tissue sections of tuberculosis patients containing granulomatous structure. Results: 11 out of 150 FFPE biopsy specimens of tuberculosis patients showed eosinophilic hyaline-rich deposits surrounding granuloma. Upon Congo red staining, these deposits exhibited characteristic apple-green birefringence under polarized light, confirming amyloid deposits. Further, upon immunohistochemical staining with anti-SAA, the amyloid enriched areas showed positive immunoreactivity. Conclusion: In this pilot study, we have shown that granuloma can be a potential site for serum amyloid A-derived amyloid formation in tuberculosis patients. Moreover, the presence of amyloid gave significant cues that granuloma might be a probable amyloid deposition start in tuberculosis patients. This study will set a stage to expand the clinical and fundamental research in the understanding of amyloid formation in granuloma underlying tuberculosis and chronic inflammatory conditions.

Keywords: amyloid, granuloma, periphery, serum amyloid A, tuberculosis

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Authors: Caroline Mendes, Mary McNamara, Orla Howe

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For chemotherapy, a drug delivery system should be able to specifically target cancer cells and deliver the therapeutic dose without affecting normal cells. Folate receptors (FR) can be considered key targets since they are commonly over-expressed in cancer cells and they are the molecular marker used in this study. Here, cyclodextrin (CD) has being studied as a vehicle for delivering the chemotherapeutic drug, methotrexate (MTX). CDs have the ability to form inclusion complexes, in which molecules of suitable dimensions are included within the CD cavity. In this study, β-CD has been modified using folic acid so as to specifically target the FR molecular marker. Thus, the system studied here for drug delivery consists of β-CD, folic acid and MTX (CDEnFA:MTX). Cellular uptake of folic acid is mediated with high affinity by folate receptors while the cellular uptake of antifolates, such as MTX, is mediated with high affinity by the reduced folate carriers (RFCs). This study addresses the gene (mRNA) and protein expression levels of FRs and RFCs in the cancer cell lines CaCo-2, SKOV-3, HeLa, MCF-7, A549 and the normal cell line BEAS-2B, quantified by real-time polymerase chain reaction (real-time PCR) and flow cytometry, respectively. From that, four cell lines with different levels of FRs, were chosen for cytotoxicity assays of MTX and CDEnFA:MTX using the MTT assay. Real-time PCR and flow cytometry data demonstrated that all cell lines ubiquitously express moderate levels of RFC. These experiments have also shown that levels of FR protein in CaCo-2 cells are high, while levels in SKOV-3, HeLa and MCF-7 cells are moderate. A549 and BEAS-2B cells express low levels of FR protein. FRs are highly expressed in all the cancer cell lines analysed when compared to the normal cell line BEAS-2B. The cell lines CaCo-2, MCF-7, A549 and BEAS-2B were used in the cell viability assays. 48 hours treatment with the free drug and the complex resulted in IC50 values of 93.9 µM ± 9.2 and 56.0 µM ± 4.0 for CaCo-2 for free MTX and CDEnFA:MTX respectively, 118.2 µM ± 10.8 and 97.8 µM ± 12.3 for MCF-7, 36.4 µM ± 6.9 and 75.0 µM ± 8.5 for A549 and 132.6 µM ± 12.1 and 288.1 µM ± 16.3 for BEAS-2B. These results demonstrate that MTX is more toxic towards cell lines expressing low levels of FR, such as the BEAS-2B. More importantly, these results demonstrate that the inclusion complex CDEnFA:MTX showed greater cytotoxicity than the free drug towards the high FR expressing CaCo-2 cells, indicating that it has potential to target this receptor, enhancing the specificity and the efficiency of the drug.

Keywords: cyclodextrins, cancer treatment, drug delivery, folate receptors, reduced folate carriers

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Authors: Shu-Pin Huang, Pai-Chi Teng, Hao-Han Chang, Chia-Hsin Liu, Yung-Lun Lin, Shu-Chi Wang, Hsin-Chih Yeh, Chih-Pin Chuu, Jiun-Hung Geng, Li-Hsin Chang, Wei-Chung Cheng, Chia-Yang Li

Abstract:

The emergence of immune checkpoint inhibitors (ICIs) targeting proteins like PD-1 and PD-L1 has changed the treatment paradigm of bladder cancer. However, not all patients benefit from ICIs, with some experiencing early death. There's a significant need for biomarkers associated with the PD-1 pathway in bladder cancer. Current biomarkers focus on tumor PD-L1 expression, but a more comprehensive understanding of PD-1-related biology is needed. Our study has developed a seven-gene risk score panel, employing a comprehensive bioinformatics strategy, which could serve as a potential prognostic and predictive biomarker for bladder cancer. This panel incorporates the FYN, GRAP2, TRIB3, MAP3K8, AKT3, CD274, and CD80 genes. Additionally, we examined the relationship between this panel and immune cell function, utilizing validated tools such as ESTIMATE, TIDE, and CIBERSORT. Our seven-genes panel has been found to be significantly associated with bladder cancer survival in two independent cohorts. The panel was also significantly correlated with tumor infiltration lymphocytes, immune scores, and tumor purity. These factors have been previously reported to have clinical implications on ICIs. The findings suggest the potential of a PD-1 pathway-based transcriptomic panel as a prognostic and predictive biomarker in bladder cancer, which could help optimize treatment strategies and improve patient outcomes.

Keywords: bladder cancer, programmed cell death protein 1, prognostic biomarker, immune checkpoint inhibitors, predictive biomarker

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