Search results for: protein concentrate

Authors: Victor Arokia Doss, Kuberapandian Dharaniyambigai, K. Julia Rose Mary

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Advanced Glycation End (AGE) products are the end products due to the reaction between excess reducing sugar present in diabetes and free amino group in protein lipids and nucleic acids. Thus, non-enzymic glycation of molecules such as hemoglobin, collagen, and other structurally and functionally important proteins add to the pathogenic complications such as diabetic retinopathy, neuropathy, nephropathy, vascular changes, atherosclerosis, Alzheimer's disease, rheumatoid arthritis, and chronic heart failure. The most common non-cross linking AGE, carboxymethyl lysine (CML) is formed by the oxidative breakdown of fructosyllysine, which is a product of glucose and lysine. CML is formed in a wide variety of tissues and is an index to assess the extent of glycoxidative damage. Thus we have constructed a mathematical and computational model that predicts the effect of temperature differences in vivo, on the formation of CML, which is now being considered as an important intracellular milieu. This hybrid model that had been tested for its parameter fitting and its sensitivity with available experimental data paves the way for designing novel laboratory experiments that would throw more light on the pathological formation of AGE adducts and in the pathophysiology of diabetic complications.

Keywords: advanced glycation end-products, CML, mathematical model, computational model

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Authors: Yao Huang, Hongjie Yu, Fangrong Xu, Longbiao Cai, Qiqiang He

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The Klotho gene has been found to be involved in cardiovascular health, and epigenetic mechanism has risen as good candidates to understand the role of lifestyle factors in obesity. The aim of this study was to investigate the effect of exercise intervention on the expression and DNA methylation of Klotho gene in high-fat diet induced obese mice. C57BL/6 male mice were fed a normal diet (ND) or a high-fat diet (HFD) for 12 weeks. HFD induced obese mice were divided into secondary group (SED) and exercise group (EX) randomly. The treadmill exercise was performed in EX group for 8 weeks. The expression and DNA methylation of Klotho were evaluated by Western blot, RT-PCR, and Methylation-specific PCR. Results indicated that Klotho protein and mRNA expression were significantly lower in the SED group than those in the ND and EX groups (P<0.01), whereas no significant difference, was found between ND group and EX group (P>0.05). Furthermore, mice in the ND group and SED group showed significantly lower levels of completely methylated Klotho DNA in ND group (0%) and SED group (50%) compared with the EX group (90%), and unmethylated Klotho DNA level in ND group (80%) was significantly higher than those in the SED (0%) and EX (0%) groups. These results suggested that exercise leads to increased Klotho expression and reduced Klotho DNA methylation level in HFD induced obese mice.

Keywords: DNA methylation, exercise intervention, klotho, obese mice

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Authors: Spencer Rutherford, Shadi Saleh

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War and conflict negatively impact all facets of a health system; services cease to function, resources become depleted, and any semblance of governance is lost. Following cessation of conflict, the rebuilding process includes a wide array of international and local actors. During this period, stakeholders must contend with various trade-offs, including balancing sustainable outcomes with immediate health needs, introducing health reform measures while also increasing local capacity, and reconciling external assistance with local legitimacy. Compounding these factors are additional challenges, including coordination amongst stakeholders, the re-occurrence of conflict, and ulterior motives from donors and governments, to name a few. Therefore, the present paper evaluated health system development in three post-conflict countries over a 12-year timeline. Specifically, health policies, health inputs (such infrastructure and human resources), and measures of governance, from the post-conflict periods of Afghanistan, Cambodia, and Mozambique, were assessed against health outputs and other measures. All post-conflict countries experienced similar challenges when rebuilding the health sector, including; division and competition between donors, NGOs, and local institutions; urban and rural health inequalities; and the re-occurrence of conflict. However, countries also employed unique and effective mechanisms for reconstructing their health systems, including; government engagement of the NGO and private sector; integration of competing factions into the same workforce; and collaborative planning for health policy. Based on these findings, best-practice development strategies were determined and compiled into a 12-year framework. Briefly, during the initial stage of the post-conflict period, primary stakeholders should work quickly to draft a national health strategy in collaboration with the government, and focus on managing and coordinating NGOs through performance-based partnership agreements. With this scaffolding in place, the development community can then prioritize the reconstruction of primary health care centers, increasing and retaining health workers, and horizontal integration of immunization services. The final stages should then concentrate on transferring ownership of the health system national institutions, implementing sustainable financing mechanisms, and phasing-out NGO services. Overall, these findings contribute post-conflict health system development by evaluating the process holistically and along a timeline and can be of further use by healthcare managers, policy-makers, and other health professionals.

Keywords: Afghanistan, Cambodia, health system development, health system reconstruction, Mozambique, post-conflict, state-building

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Authors: Jeremy J. Johnson

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Mediterranean herbs including rosemary (Rosmarinus officinalis) contain a variety of phytochemicals including diterpenes that possess extensive biological activity. Applications of diterpenes, including the more abundant forms carnosol and carnosic acid, have been shown to possess anti-cancer, anti-inflammatory, anti-oxidant, and anti-proliferation properties. To confirm these properties, we have evaluated rosemary extract and selected diterpenes for biological activity in cancer and inflammatory models. Our preliminary data have revealed that select diterpenes can disrupt androgen receptor functionality in prostate and breast cancer cells. This property is unique among natural products for hormone-responsive cancers. The second area of interest has been evaluating rosemary extract and selected diterpenes for activation of sestrin-2, an antioxidant protein, in colon cancer cells. A combination of in vitro and in vivo approaches have been utilized to characterize the activity of rosemary diterpenes in rosemary. Taken together, these results suggest that phytochemicals found in rosemary have distinct pharmacological actions for disrupting cell-signaling pathways in cancer and inflammatory bowel disease.

Keywords: rosemary, diterpene, cancer, inflammation

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Authors: Sana Abbasa, Shahzad Bhattiab, Nadir Khan

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Sargassum siliquastrum is brown seaweed with traditional claims for some medicinal properties. This research was done to investigate the methanol extract of S. siliquastrum for antiproliferative activity against human cervical cancer cell line, HeLa and its mode of cell death. From methylene blue assay, S. siliquastrum exhibited antiproliferative activity on HeLa cells with IC50 of 3.87 µg/ml without affecting non-malignant cells. Phase contrast microscopy indicated the confluency reduction in HeLa cells and changes on the cell shape. Nuclear staining with Hoechst 33258 displayed the formation of apoptotic bodies and fragmented nuclei. S. siliquastrum also induced early apoptosis event in HeLa cells as confirmed by FITC-Annexin V/propidium iodide staining by flow cytometry analysis. Cell cycle analysis indicated growth arrest of HeLa cells at G1/S phase. Protein study by flow cytometry indicated the increment of p53, slight increase of Bax and unchanged level of Bcl-2. In conclusion, S. siliquastrum demonstrated an antiproliferative activity in HeLa cell by inducing G1/S cell cycle arrest via p53-mediated pathway.

Keywords: sargassum siliquastrum, cervical cancer, P53, antiproleferation

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Authors: E. Izsó, M. Bartalné-Berceli, Sz. Gergely, A. Salgó

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The aims of this investigation is to elaborate near-infrared methods for testing and recognition of chemical components and quality in “Pannon wheat” allied (i.e. true to variety or variety identified) milling fractions as well as to develop spectroscopic methods following the milling processes and evaluate the stability of the milling technology by different types of milling products and according to sampling times, respectively. This wheat categories produced under industrial conditions where samples were collected versus sampling time and maximum or minimum yields. The changes of the main chemical components (such as starch, protein, lipid) and physical properties of fractions (particle size) were analysed by dispersive spectrophotometers using visible (VIS) and near-infrared (NIR) regions of the electromagnetic radiation. Close correlation were obtained between the data of spectroscopic measurement techniques processed by various chemometric methods (e.g. principal component analysis (PCA), cluster analysis (CA) and operation condition of milling technology. Its obvious that NIR methods are able to detect the deviation of the yield parameters and differences of the sampling times by a wide variety of fractions, respectively. NIR technology can be used in the sensitive monitoring of milling technology.

Keywords: near infrared spectroscopy, wheat categories, milling process, monitoring

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Authors: John O. Jacob, Abdullahi Mann, Olanrewaju I. Adeshina, Mohammed M. Ndamitso

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Strychnos spinosa, Detarium microcarpum, Diospyros mespiliformis, Dialium guineese and Gardenia ternifolia are some of the wild fruits consume in the villages around Minna, Niger State. This investigation was conducted to assess the nutritional potentials of these fruits both for human consumption and for possible application in animal feed formulations. Standard analytical methods were employed in the determination of the various nutritional parameters. The proximate analysis results showed that the moisture contents ranged between (6.17-10.70%); crude fat (2.04-8.85%); crude protein (5.16-6.80%); crude fibre (7.23-19.65%); Ash (3.46-5.56%); carbohydrate (57.77-69.79%); energy value (284.49-407 kcal/mg); Vitamin C (7.2-39.93 mg/100g). The mineral analysis shows that the selected wild fruits could contribute considerable amount of both micro and macro elements to human nutrition potassium, sodium and calcium range between; potassium (343.27-764.71%); sodium (155.04-348.44%); calcium (52.47-101%). The macro element for the fruits pulp were in the order K>Na>Mg>Ca, hence, they could be included in diet to supplement daily nutrient requirement and in animal feed formulations. The domestication of these fruits is also encouraged.

Keywords: mineral, micro-elements, macro-elements, feed suppleme

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Authors: Sara Saboonian, Pierre Filion

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There are two distinct planning approaches attempting to transform the North American suburb so as to reduce its adverse environmental impacts. The first one, the recentralization approach, proposes intensification, multi-functionality and more reliance on public transit and walking. It thus offers an alternative to the prevailing low-density, spatial specialization and automobile dependence of the North American suburb. The second approach concentrates instead on the provision of green infrastructure, which rely on natural systems rather than on highly engineered solutions to deal with the infrastructure needs of suburban areas. There are tensions between these two approaches as recentralization generally overlooks green infrastructure, which can be space consuming (as in the case of water retention systems), and thus conflicts with the intensification goals of recentralization. The research investigates three Canadian planned suburban centres in the Greater Toronto Area, where recentralization is the current planning practice, despite rising awareness of the benefits of green infrastructure. Methods include reviewing the literature on green infrastructure planning, a critical analysis of the Ontario provincial plans for recentralization, surveying residents’ preferences regarding alternative suburban development models, and interviewing officials who deal with the local planning of the three centres. The case studies expose the difficulties in creating planned suburban centres that accommodate green infrastructure while adhering to recentralization principles. Until now, planners have been mostly focussed on recentralization at the expense of green infrastructure. In this context, the frequent lack of compatibility between recentralization and the space requirements of green infrastructure explains the limited presence of such infrastructures in planned suburban centres. Finally, while much attention has been given in the planning discourse to the economic and lifestyle benefits of recentralization, much less has been made of the wide range of advantages of green infrastructure, which explains limited public mobilization over the development of green infrastructure networks. The paper will concentrate on ways of combining recentralization with green infrastructure strategies and identify the aspects of the two approaches that are most compatible with each other. The outcome of such blending will marry high density, public-transit oriented developments, which generate walkability and street-level animation, with the presence of green space, naturalized settings and reliance on renewable energy. The paper will advance a planning framework that will fuse green infrastructure with recentralization, thus ensuring the achievement of higher density and reduced reliance on the car along with the provision of critical ecosystem services throughout cities. This will support and enhance the objectives of both green infrastructure and recentralization.

Keywords: environmental-based planning, green infrastructure, multi-functionality, recentralization

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Authors: Emmanuel Okine Neokye, Xiuquan Wang, Krishna K. Thakur, Pedro Quijon, Rana Ali Nawaz, , Sana Basheer

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Oysters are enriched with high-quality protein and are widely known for their exquisite taste. The production of oysters plays an important role in the local economies of coastal communities in many countries, including Atlantic Canada, because of their high economic value. However, because of the changing climatic conditions in recent years, oyster aquaculture faces potentially negative impacts, such as increasing water acidification, rising water temperatures, high salinity, invasive species, algal blooms, and other environmental factors. Although a few isolated effects of climate change on oyster aquaculture have been reported in recent years, it is not well understood how climate change will affect oyster aquaculture from a systematic perspective. In the first part of this study, we present a systematic review of the impacts of climate change and some key environmental factors affecting oyster production on a global scale. The study also identifies knowledge gaps and challenges. In addition, we present key research directions that will facilitate future investigations.

Keywords: climate change, oyster production, oyster aquaculture, greenhouse gases

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Authors: Laleh Behboudi Tabrizi, Melika Naserzare

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Accumulating experimental and epidemiologic data smoker individuals are more prone to systemic inflammation than non-smokers. In this study we aimed to determine serum TNF-α and C-reactive protein (CRP) as pro-inflammatory cytokines in response to 3 months aerobic training in smoker men. A total 30 middle-aged healthy smokers selected for participate in this study and were divided into either control or exercise groups. The subjects in exercise group were completed a 3 months aerobic training program for 3 sessions per week at 60 – 80 % of maximal heart rate. Those in control group did nit participated in exercise training. Pre and post-training of CRP and TNF-α were measured in two groups. Student’s t-tests for paired samples were performed to determine whether there were signigcant within-group changes in the outcomes. P value of <0.05 was accepted as significant. No significant differences were found in anthropometrical and biochemical markers between two groups at baseline. Aerobic training program resulted in a significant decrease in anthropometrical markers and serum TNF-α but not in serum CRP in exercise group. All variables remained without changes in control groups. Based on these finding, it is concluded that aerobic training can be improve inflammatory cytokine with emphasis on TNF-α in smokers.

Keywords: cigarette, cytokine, chronic training, inflammation

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Authors: Maryam Hashemi, Nematollah Razmi, Rasool Madani

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M. paratuberculosis is a slow growing mycobactin dependent mycobacterial species known to be the causative agent of Johne’s disease in all species of domestic ruminants worldwide. JD is characterized by gradual weight loss; decreased milk production. Excretion of the organism may occur for prolonged periods (1 to 2.5 years) before the onset of clinical disease. In recent years, researchers focus on identification a specific antigen of MAP to use in diagnosis test and preparation of effective vaccine. In this paper, for production of polyclonal antibody against proteins of Mycobacterium avium paratuberculosis cell wall a rabbit immunization at a certain time period with antigen. After immunization of the animal, blood samples were collected from the rabbit for producing enriched serum. Antibodies were purified with ion exchange chromatography. For exact measurement of interaction, western blotting test was used and as it is demonstrated in the study, sharp bands appear in nitrocellulose paper and specific bands were 50 and 150 KD molecular weight. These were indicating immunodominant proteins.

Keywords: immunodominant, paratuberculosis, Western blotting, cell wall proteins, protein purification

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Authors: Mehran Bakhshi, Mohammad Raouf Hosseini, Mohammadhosein Rahimi

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The cadmium sulfide nanoparticles were synthesized from the nickel-cadmium cake of a hydrometallurgical zinc producing plant and sodium sulfide as Cd²⁺ and S^-2 sources, respectively. Also, the synthesis process was performed by using the secretions of Bacillus licheniformis as bio-surfactant. Initially, in order to obtain a cadmium rich solution, two following steps were carried out: 1) Alkaline leaching for the removal of zinc oxide from the cake, and 2) acidic leaching to dissolve cadmium from the remained solid residue. Afterward, the obtained CdSO₄ solution was used for the nanoparticle biosynthesis. Nanoparticles were characterized by the energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) to confirm the formation of CdS crystals with cubic structure. Also, transmission electron microscopy (TEM) was applied to determine the particle sizes which were in 2-10 nm range. Moreover, the presence of the protein containing bio-surfactants was approved by using infrared analysis (FTIR). In addition, the absorbance below 400 nm confirms quantum particles’ size. Finally, it was shown that valuable CdS quantum dots could be obtained from the industrial waste products via environment-friendly biological approaches.

Keywords: biosynthesis, cadmium cake, cadmium sulfide, nanoparticle, zinc smelter

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Authors: M. Sorahi Nobar, V. Niknam, H. Ebrahimzadeh, H. Soltanloo

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Fusarium graminearum is one of the most destructive crop diseases in the world. Infection occurs during the flowering period in warm and humid conditions. It causes reduction in yield. Moreover, harvested grain is often contaminated with mycotoxins and its acetylated derivatives. Fusarium mycotoxines are potent inhibitor of protein synthesis, and thereby presents hazards for both human and animal health. A rapid production of reactive oxygen intermediates, primarily superoxide and hydrogen peroxide at the site of attempted infection considered as key feature underlying successful pathogen recognition. Here, we compared the time course activity of superoxide dismutase (SOD) as a first line of defenses against ROS- induced oxidative burst between FHB- resistant Sumai3 and susceptible Falat at 48, 96 and 144 hours after infection. Our results showed that Sumai3 SOD activity increased with time and reached the highest-level 4 days after infection while in susceptible cultivar Falat, SOD activity decreased during the first 96 h. after infection. Decreased was followed by an increased at 6 days after infection. According to our results rapid induction of SOD activity in resistant cultivar may play an important role in resistance against FHB in wheat.

Keywords: Fusarium graminearum, mycotoxins, resistant cultivar, superoxide dismutase

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Authors: Marcin Kruszewski, Barbara Sochanowicz, Sylwia Męczyńska-Wielgosz, Maria Wojewódzka, Lucyna Kapka-Skrzypczak

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Metallic NPs are widely used in a number of applications in industry, science and medicine. Among metallic NPs foreseen to be widely used in medicine are gold nanoparticles (AuNPs) due to their low toxicity, and silver NPs (AgNPs) due to their strong antimicrobial activity. In this study, we compared an effect of AgNPs and gold NPs (AuNPs) on the formation of DNA damage and global DNA methylation and in A2780 and 4T1 cell lines, widely used models of human ovarian carcinoma and murine mammary carcinoma, respectively. The cells were treated with AgNPs coated with citrate (AgNPs(cit) or PEG (AgNPs(PEG), or AuNPs. A global DNA methylation was investigated with ELISA, whereas the formation of DNA damage was investigated by a comet +/- FPG. AgNPs decreased global DNA methylation and increased the formation of DNA lesions in both cell lines. The effect was dependent on the type of NPs used, it's coating, and cell line used. In conclusion, the epigenetic and genotoxic effects of NPs strongly depends on NP nature and cellular context. Epigenetic changes observed upon the action of AgNPs may play a crucial role in NPs-induced changes in protein expression.

Keywords: DNA damage, gold nanoparticles, methylation, silver nanoparticles

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Authors: Xiaoqiang Fu, Baihao Li

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In 1895, the failure of Sino-Japanese prompted the trend of comprehensive and systematic study of western pattern in China. In urban planning and construction, urban reform movement sprang up slowly, which aimed at renovating and reconstructing the traditional cities into modern cities similar to the concessions. During the movement, Chinese traditional city initiated a process of modern urban planning for its modernization. Meanwhile, the traditional planning morphology and system started to disintegrate, on the contrary, western form and technology had become the paradigm. Therefore, the improvement of existing cities had become the prototype of urban planning of early modern China. Currently, researches of the movement mainly concentrate on large cities, concessions, railway hub cities and some special cities resembling those. However, the systematic research about the large number of traditional small and medium-sized cities is still blank, up to now. This paper takes the improvement constructions of small and medium-sized cities in Southern region of Jiangsu Province as the research object. First of all, the criteria of small and medium-sized cities are based on the administrative levels of general office and cities at the county level. Secondly, the suitability of taking the Southern Jiangsu as the research object. The southern area of Jiangsu province called Southern Jiangsu for short, was the most economically developed region in Jiangsu, and also one of the most economically developed and the highest urbanization regions in China. As the most developed agricultural areas in ancient China, Southern Jiangsu formed a large number of traditional small and medium-sized cities. In early modern times, with the help of the Shanghai economic radiation, geographical advantage and powerful economic foundation, Southern Jiangsu became an important birthplace of Chinese national industry. Furthermore, the strong business atmosphere promoted the widespread urban improvement practices, which were incomparable of other regions. Meanwhile, the demonstration of Shanghai, Zhenjiang, Suzhou and other port cities became the improvement pattern of small and medium-sized city in Southern Jiangsu. This paper analyzes the reform movement of the small and medium-sized cities in Southern Jiangsu (1895-1927), including the subjects, objects, laws, technologies and the influence factors of politic and society, etc. At last, this paper reveals the formation mechanism and characteristics of urban improvement movement in early modern China. According to the paper, the improvement of small-medium city was a kind of gestation of the local city planning culture in early modern China，with a fusion of introduction and endophytism.

Keywords: early modern China, improvement of small-medium city, southern region of Jiangsu province, urban planning history of China

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Authors: Ikele Chika Bright, Mgbenka Bernard Obialo, Ikele Chioma Faith

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Icthyophthiriasis is a prevalent protozoan (ectoparasite) mostly affecting cultured and aquarium fishes. The majority of the chemotherapeutants lack efficacy for completely eliminating Ich parasite without affecting the environment and they are not safe for human health. The present work is focused on the evaluating different immersion treatments of African catfish (Clarias gariepinus) infected with ichthyophthiriasis and treated with a non-chemical and environmental friendly parasiticides Moringa oleifera. A total number of 800 apparently healthy parasites free (examined) post juvenile catfish were obtained from a reputable farm, disinfected with potassium permanganate in a quarantine tank to remove any possible external parasites. The fish were further challenged with approximately 44,000 infective stages of theronts which were obtained through serial passages by cohabitation. Seven groups (A-G) of post Juvenile were used for the experiment which was carried out into three stages; Dips (60minutes), short term treatment (24-96h) and prolong bath treatment (0-15 days). The concentrations selected were dependent on the outcome of the LC50 of the plant material from which dose-dependent factors were used to select various concentrations of the treatment. In Dips treatment, group D-G were treated with 1,500mg/L, 2500mg/L., 3500mg/L and 4500mg/L, short-term treatment was treated with 150mg/L, 250mg/L, 350mg/L and 450mg/L and prolong bath was treated with 15mg/L, 25mg/L, 35mg/L and 45mg/L of the plant extract whereas group A, B and C were normal control, Ich- infested not treated and Ich- infested treated with standard drug (Acriflavin), respectively. The various types of treatment applied with corresponding concentrations showed almost complete elimination of the adult parasites (trophonts) both in the gills and the body smear, thereby making M. oleifera a potential parasiticides. There were serious pathological alterations in the skin and gills which are usually the main point for Ich parasites invasion but no significant morphological characteristics was noted among the treated groups subjected to different immersion treatment patterns. Epitheliocystis, aneurysm, oedema, hemorrhage, and localization of the adult parasite in the gills were the overall common observations made in the gills whereas degeneration of muscle fibre, dermatitis, hemorrhage, oedema, abscess formation and keratinisation were observed in the skin. However, there are no pathological changes in the control group. Moreover, biochemical parameters such as urea, creatinine, albumin., globulin, total protein, ALT, AST), blood chemistry (sodium, chloride, potassium, bicarbonate), antioxidants (CAT, SOD, GPx, LPO), enzymatic activities (myeloperoxidase, thioreadoxin reductase), Inflammatory response (C-reactive protein), Stress markers (lactate dehydrogenase), heamatological parameters (RBC, PCV, WBC, HB and differential count), lipid profile (total cholesterol, tryglycerides , high density lipoprotein and low density lipoprotein) all showed various significant (P<0.05) and no significant (P>0.05) responses among the Ich-infested fish treated under three immersion treatments. It is suggested that M. oleifera may serve as an alternatives to chemotherapeutants for control of Ichthyophthiriasis in African catfish Clarias gariepinus.

Keywords: Icthyophthirius multifilis, immersion treatment, pathophysiology, African catfish

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Authors: Fabio Fabris, Alex A. Freitas

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Hierarchical classification is a special type of classification task where the class labels are organised into a hierarchy, with more generic class labels being ancestors of more specific ones. Meta-learning for classification-algorithm recommendation consists of recommending to the user a classification algorithm, from a pool of candidate algorithms, for a dataset, based on the past performance of the candidate algorithms in other datasets. Meta-learning is normally used in conventional, non-hierarchical classification. By contrast, this paper proposes a meta-learning approach for more challenging task of hierarchical classification, and evaluates it in a large number of bioinformatics datasets. Hierarchical classification is especially relevant for bioinformatics problems, as protein and gene functions tend to be organised into a hierarchy of class labels. This work proposes meta-learning approach for recommending the best hierarchical classification algorithm to a hierarchical classification dataset. This work’s contributions are: 1) proposing an algorithm for splitting hierarchical datasets into new datasets to increase the number of meta-instances, 2) proposing meta-features for hierarchical classification, and 3) interpreting decision-tree meta-models for hierarchical classification algorithm recommendation.

Keywords: algorithm recommendation, meta-learning, bioinformatics, hierarchical classification

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Authors: N. Othman, J. Ujang, M. N. Ismail, R. Noordin, B. H. Lim

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Amoebiasis is caused by the Entamoeba histolytica and still endemic in many parts of the tropical region, worldwide. Currently, there is no available vaccine against amoebiasis. Hence, there is an urgent need to develop a vaccine. The excretory secretory antigen (ESA) of E. histolytica is a suitable biomarker for the vaccine candidate since it can modulate the host immune response. Hence, the objective of this study is to identify the proteome of the ESA towards finding suitable biomarker for the vaccine candidate. The non-gel based and gel-based proteomics analyses were performed to identify proteins. Two kinds of mass spectrometry with different ionization systems were utilized i.e. LC-MS/MS (ESI) and MALDI-TOF/TOF. Then, the functional proteins classification analysis was performed using PANTHER software. Combination of the LC -MS/MS for the non-gel based and MALDI-TOF/TOF for the gel-based approaches identified a total of 273 proteins from the ESA. Both systems identified 29 similar proteins whereby 239 and 5 more proteins were identified by LC-MS/MS and MALDI-TOF/TOF, respectively. Functional classification analysis showed the majority of proteins involved in the metabolic process (24%), primary metabolic process (19%) and protein metabolic process (10%). Thus, this study has revealed the proteome the E. histolytica ESA and the identified proteins merit further investigations as a vaccine candidate.

Keywords: E. histolytica, ESA, proteomics, biomarker

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Authors: Saleh Gorji, Sahil Gulati, Ana Pakzad

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Continuous electron diffraction tomography, also known as microcrystal electron diffraction (MicroED) or three-dimensional electron diffraction (3DED), is a powerful technique, which in combination with cryo-electron microscopy (cryo-ED), can provide atomic-scale 3D information about the crystal structure and composition of different classes of crystalline materials such as proteins, peptides, and small molecules. Unlike the well-established X-ray crystallography method, 3DED does not require large single crystals and can collect accurate electron diffraction data from crystals as small as 50 – 100 nm. This is a critical advantage as growing larger crystals, as required by X-ray crystallography methods, is often very difficult, time-consuming, and expensive. In most cases, specimens studied via 3DED method are electron beam sensitive, which means there is a limitation on the maximum amount of electron dose one can use to collect the required data for a high-resolution structure determination. Therefore, collecting data using a conventional scintillator-based fiber coupled camera brings additional challenges. This is because of the inherent noise introduced during the electron-to-photon conversion in the scintillator and transfer of light via the fibers to the sensor, which results in a poor signal-to-noise ratio and requires a relatively higher and commonly specimen-damaging electron dose rates, especially for protein crystals. As in other cryo-EM techniques, damage to the specimen can be mitigated if a direct detection camera is used which provides a high signal-to-noise ratio at low electron doses. In this work, we have used two classes of such detectors from Gatan, namely the K3® camera (a monolithic active pixel sensor) and Stela™ (that utilizes DECTRIS hybrid-pixel technology), to address this problem. The K3 is an electron counting detector optimized for low-dose applications (like structural biology cryo-EM), and Stela is also a counting electron detector but optimized for diffraction applications with high speed and high dynamic range. Lastly, data collection workflows, including crystal screening, microscope optics setup (for imaging and diffraction), stage height adjustment at each crystal position, and tomogram acquisition, can be one of the other challenges of the 3DED technique. Traditionally this has been all done manually or in a partly automated fashion using open-source software and scripting, requiring long hours on the microscope (extra cost) and extensive user interaction with the system. We have recently introduced Latitude® D in DigitalMicrograph® software, which is compatible with all pre- and post-energy-filter Gatan cameras and enables 3DED data acquisition in an automated and optimized fashion. Higher quality 3DED data enables structure determination with higher confidence, while automated workflows allow these to be completed considerably faster than before. Using multiple examples, this work will demonstrate how to direct detection electron counting cameras enhance 3DED results (3 to better than 1 Angstrom) for protein and small molecule structure determination. We will also show how Latitude D software facilitates collecting such data in an integrated and fully automated user interface.

Keywords: continuous electron diffraction tomography, direct detection, diffraction, Latitude D, Digitalmicrograph, proteins, small molecules

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Authors: Naiduwadura Ivon Upekala De Silva, Nathan Lehman, Talia Fargason, Trenton Paul, Zihan Zhang, Jun Zhang

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SRSF1 governs splicing of over 1,500 mRNA transcripts. SRSF1 contains two RNA-recognition motifs (RRMs) and a C-terminal Arg/Ser-rich region (RS). It has been thought that SRSF1 RRMs exclusively recognize single-stranded exonic splicing enhancers, while RS lacks RNA-binding specificity. With our success in solving the insolubility problem of SRSF1, we can explore the unknown RNA-binding landscape of SRSF1. We find that SRSF1 RS prefers purine over pyrimidine. Moreover, SRSF1 binds to the G-quadruplex (GQ) from the ARPC2 mRNA, with both RRMs and RS being crucial. Our binding assays show that the traditional RNA-binding sites on the RRM tandem and the Arg in RS are responsible for GQ binding. Interestingly, our FRET and circular dichroism data reveal that SRSF1 unfolds the ARPC2 GQ, with RS leading unfolding and RRMs aiding. Our saturation transfer difference NMR results discover that Arg residues in SRSF1 RS interact with the guanine base but with other nucleobases, underscoring the uniqueness of the Arg/guanine interaction. Our luciferase assays confirm that SRSF1 can alleviate the inhibitory effect of GQ on gene expression in the cell. Given the prevalence of RNA GQ and SR proteins, our findings unveil unexplored SR protein functions with broad implications in RNA splicing and translation.

Keywords: SR, SRSF!, RNA G-quadruplex, unfolding, RNA binding

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Authors: Eman Alzahrani

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Efficient extraction of proteins by removing interfering materials is necessary in proteomics, since most instruments cannot handle such contaminated sample matrices directly. In this study, chitosan-coated magnetic nanoparticles (CS-MNPs) for purification of myoglobin were successfully fabricated. First, chitosan (CS) was prepared by a deacetylation reaction during its extraction from shrimp-shell waste. Second, magnetic nanoparticles (MNPs) were synthesised, using the coprecipitation method, from aqueous Fe2+ and Fe3+ salt solutions by the addition of a base under an inert atmosphere, followed by modification of the surface of MNPs with chitosan. The morphology of the formed nanoparticles, which were about 23 nm in average diameter, was observed by transmission electron microscopy (TEM). In addition, nanoparticles were characterised using X-ray diffraction patterns (XRD), which showed the naked magnetic nanoparticles have a spinel structure and the surface modification did not result in phase change of the Fe3O4. The coating of MNPs was also demonstrated by scanning electron microscopy (SEM) analysis, energy dispersive analysis of X-ray spectroscopy (EDAX), and Fourier transform infrared (FT-IR) spectroscopy. The adsorption behaviour of MNPs and CS-MNPs towards myoglobin was investigated. It was found that the difference in adsorption capacity between MNPs and CS-MNPs was larger for CS-MNPs. This result makes CS-MNPs good adsorbents and attractive for using in protein extraction from biological samples.

Keywords: chitosan, magnetic nanoparticles, coprecipitation, adsorption

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Authors: Izdihar Ismail, Alona C. Linatoc, Maryati Mohamed

Abstract:

The tropical rainforest of Malaysia is known for its rich biological diversity and high endemicity. The potential for these forests to hold the cure for many diseases and illnesses is high and much is yet to be discovered. This study explores the richness of the tropical rainforest of Endau-Rompin National Park in Johor, Malaysia in search of plants traditionally used by the indigenous people in the treatment of malaria and malaria-like symptoms. Seven species of plants were evaluated and tested for antiplasmodial activities. Different plant parts were subjected to methanolic and aqueous extractions. A total of 24 extracts were evaluated by histidine-rich protein II (HRP2) assay against K1 strain of Plasmodium falciparum chloroquine-resistant. Ten extracts showed significant inhibition of the growth of P. falciparum. Phytochemical screening of the same extracts revealed the presence of alkaloids, flavonoids, terpenoids and anthraquinones. This study affirms that tropical rainforests may still hold undiscovered cures for many diseases and illnesses that have inflicted millions of people worldwide. The species studied herein have not known to have been studied elsewhere before.

Keywords: Endau-Rompin, malaria, Malaysia, tropical rainforest, traditional knowledge

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Authors: Muhammad Asif Rasheed

Abstract:

Newcastle disease virus (NDV), an avian orthoavulavirus, is a causative agent of Newcastle disease named (NDV) and can cause even the epidemics when the disease is not treated. Previously several vaccines based on attenuated and inactivated viruses have been reported, which are rendered useless with the passage of time due to versatile changes in viral genome. Therefore, we aimed to develop an effective multi-epitope vaccine against the haemagglutinin neuraminidase (HN) protein of 26 NDV strains from Pakistan through a modern immunoinformatic approaches. As a result, a vaccine chimaera was constructed by combining T-cell and B-cell epitopes with the appropriate linkers and adjuvant. The designed vaccine was highly immunogenic, non-allergen, and antigenic; therefore, the potential 3D-structureof multi epitope vaccine was constructed, refined, and validated. A molecular docking study of a multiepitope vaccine candidate with the chicken Toll-like receptor-4 indicated successful binding. An In silico immunological simulation was used to evaluate the candidate vaccine's ability to elicit an effective immune response. According to the computational studies, the proposed multiepitope vaccine is physically stable and may induce immune responses, whichsuggested it a strong candidate against 26 Newcastle disease virus strains from Pakistan. A wet lab study is under process to confirm the results.

Keywords: epitopes, newcastle disease virus, paramyxovirus virus, vaccine

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Authors: Weili Shao, Qian Wang, Jianxin He

Abstract:

Recently, the applications of graphene oxide in fabricating scaffolds for bone tissue engineering have been received extensive concern. In this work, poly l-lactic/graphene oxide composite nanofibers were successfully fabricated by electrospinning. The morphology structure, porosity and mechanical properties of the composite nanofibers were characterized using different techniques. And mouse mesenchymal stem cells were cultured on the composite nanofiber scaffolds to assess their suitability for bone tissue engineering. The results indicated that the composite nanofiber scaffolds had finer fiber diameter and higher porosity as compared with pure poly l-lactic nanofibers. Furthermore, incorporation of graphene oxide into the poly l-lactic nanofibers increased protein adsorptivity, boosted the Young’s modulus and tensile strength by nearly 4.2-fold and 3.5-fold, respectively, and significantly enhanced adhesion, proliferation, and osteogenesis in mouse mesenchymal stem cells. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering.

Keywords: poly l-lactic, graphene oxide, osteogenesis, bone tissue engineering

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Authors: Hammed Tanimowo Aiyelabegan, Oluchukwu Franklin Aladi, Mutiu Adewumi Alabi, Raliat Abimbola Aladodo, Emmanuel Oladipupo Ajani, Abdulganiyu Giwa, Esther Owolabi

Abstract:

The study aimed to evaluate the inhibitory activities of ligands from Enantia chlorantha stem bark on α-amylase and α-glucosidase. In silico pharmacokinetic properties and docking scores were employed to analyse the inhibition using SwissADME and Autodock4.2, respectively. Results revealed that drug-likeness, pharmacokinetics and bioavailability radar of all the ligands except jatrorrhizine and acarbose falls within the radar according to the Lipinski rule of 5. The binding energies of the protein-ligand interactions also show that the ligand fits into the active site. The results obtained from this study show that the chemical constituents from Enantia chlorantha stem bark may bring about positive physiological changes in a patient suffering from diabetes mellitus. Further in vitro studies on diabetes cell lines and in vivo studies on the animal may validate these compounds for diabetes treatment. These phytoconstituents could help in the development of novel anti-diabetic molecules.

Keywords: diabetes mellitus, ?-amylase, ?-glucosidase, in silico, Enantia chlorantha stem bark

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Authors: Nnennaya Samuel Okoro

Abstract:

An experiment was conducted to determine the growth performance, blood parameters and reproductive characteristics of 8-week old male weaner rabbits fed 20% boiled pigeon pea seed meal (PPSM). The study lasted for 16 weeks. Results showed that haematological parameters of the two groups of rabbit bucks were not significantly affected (P>0.05) by the treatment, meaning that the PPSM was adequate for maintaining the blood parameters at the normal levels. The 20% boiled PPSM significantly affected (P<0.05) serum Alanine Aminotransferase (ALT) (67.72±2.5 I.U/I) more than the ALT (57.50±2.02 I.U/I) of the control, which is an indication of liver problem. The globulin level (3.00 ± 0.23 g/d) of the 20% boiled PPSM group was significantly higher than that of the control (2.60±0.06 g/dl), indicating that the test diet did not alter protein metabolism in the rabbits. Boiled pigeon pea seed meal supported organ weight and testicular development in rabbit bucks, suggesting that boiling reduced the level of the anti-nutritional factors in pigeon pea seed meal. Thus, 20% boiled pigeon pea can be included in diets of rabbits without adverse effect on blood parameters and internal organs characteristics.

Keywords: haematology, internal organs, pigeon pea, rabbits, serum biochemistry

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Authors: Chin-Yuan Hsieh, Wei-Chun Lu, Yu-Hong Zeng

Abstract:

The situation of a group of fish die is frequently found due to the fish disease caused by the deterioration of aquaculture water quality. The toxic ammonia is produced by animals as a byproduct of protein. The system is designed by the smart sensor technology and developed by the mathematical model to monitor the water parameters 24 hours a day and predict the relationship among twelve water quality parameters for monitoring the water quality in aquaculture. All data measured are stored in cloud server. In productive ponds, the daytime pH may be high enough to be lethal to the fish. The sudden change of the aquaculture conditions often results in the increase of PH value of water, lack of oxygen dissolving content, water quality deterioration and yield reduction. From the real measurement, the system can send the message to user’s smartphone successfully on the bad conditions of water quality. From the data comparisons between measurement and model simulation in fish aquaculture site, the difference of parameters is less than 2% and the correlation coefficient is at least 98.34%. The solubility rate of oxygen decreases exponentially with the elevation of water temperature. The correlation coefficient is 98.98%.

Keywords: aquaculture, sensor, ammonia, dissolved oxygen

Procedia PDF Downloads 281

Authors: Wen-Yu Lin, Yi-Ching Chung, Tzyy-Rong Jinn

Abstract:

It is well known that enterovirus 71 (EV71) causes recurring outbreaks of hand, foot and mouth disease (HFMD) and encephalitis leading to complications or death in young children. And, several HFMD of EV71 with high mortalities occurred in Asia countries, such as Malaysia (1997), Taiwan (1998) and China (2008). Thus, more effective antiviral drugs are needed to prevent or reduce EV71-related complications. As reported, interferon-α protects mice from lethal EV71 challenge by the modulation of innate immunity and then degrade enterovirus protease 3Cᵖʳᵒ. On the other side, pleconaril by targeting enterovirus VP1 protein and then block virus entry and attachment. Thus, the aim of this study was to evaluate the synergistic antiviral activity of interferon-α and pleconaril against enterovirus 71 infection. In a preliminary study showed that pleconaril at concentrations of 50, 100 and 300 µg/mL reduced EV71-induced CPE to 52.0 ± 2.5%, 40.2 ± 3.5% and 26.5 ± 1.5%, respectively, of that of the EV71-infected RD control cells (taken as 100%). Notably, 1000 IU/mL of interferon-α in combination with pleconaril at concentrations of 50, 100 and 300µg/mL suppressed EV71-induced CPE by 30.2 ± 3.8%, 16.5 ± 1.3% and 2.8 ± 2.0%, respectively, of that of the pleconaril alone treated with the infected RD cells. These results indicated that interferon-α 1000 IU/mL combination with pleconaril (50, 100 and 300µg/mL) inhibited EV71-induced CPE more effectively than treated with pleconaril alone in the infected RD cells.

Keywords: enterovirus 71, interferon-α, pleconaril, RD cells

Procedia PDF Downloads 139

Authors: Ramandeep Kaur, Kamaljit Kaur, Preeti Ahluwalia, Poonam A. Sachdev

Abstract:

Bread is an ideal vehicle to impart bioactive compounds to the consumers in a convenient manner. This study evaluated bread enriched with red sweet pepper powder (RSP) at 2, 4, 6, 8, 10% and compared to control bread (without RSP). The bread crumbs were assayed for bioactive, physical, nutritional, textural, color, and sensory properties. Bread supplemented with RSP improved its color, nutritional, and bioactive properties. The low moisture content and increased hardness were observed at higher levels of RSP. Color intensity (expressed as L*, a*, b* values) of bread with 2 and 4% RSP were lower than those of high levels, and the same trend was observed for protein, fibre and ash content of bread. Significant (p < 0.05) increases were recorded for bioactive compounds such as total phenols (0.145 to 235 mg GAE/g), antioxidant activity (56% to 78%) and flavonoids (0.112 to 0.379 mg/g) as the level of powder increased. Bread enriched with 8% RSP showed improved sensory profile as compared to control, whereas a further increase in RSP decreased the sensory and textural properties. Thus, RSP act as a natural colorant and functional food that enhanced the functional and nutritional properties of bread and can be used to customize bread for specific health needs.

Keywords: breads, bioactive compounds, red sweet pepper powder, sensory scores

Procedia PDF Downloads 156

Authors: Muhammad Ali Syed, Arshad Saleem Bhatti, Chen-zhong Li, Habib Ali Bokhari

Abstract:

Surface plasmon resonance (SPR) biosensors have emerged as a promising technique for bioanalysis as well as microbial detection and identification. Real time, sensitive, cost effective, and label free detection of biomolecules from complex samples is required for early and accurate diagnosis of infectious diseases. Like many other types of optical techniques, SPR biosensors may also be successfully utilized for microbial detection for accurate, point of care, and rapid results. In the present study, we have utilized a commercially available automated SPR biosensor of BI company to study the microbial detection form water samples spiked with different concentration of Staphylococcus aureus bacterial cells. The gold thin film sensor surface was functionalized to react with proteins such as protein G, which was used for directed immobilization of monoclonal antibodies against Staphylococcus aureus. The results of our work reveal that this immunosensor can be used to detect very small number of bacterial cells with higher sensitivity and specificity. In our case 10^3 cells/ml of water have been successfully detected. Therefore, it may be concluded that this technique has a strong potential to be used in microbial detection and identification.

Keywords: surface plasmon resonance (SPR), Staphylococcus aureus, biosensors, microbial detection

Procedia PDF Downloads 473