Search results for: protein conformation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2415

Search results for: protein conformation

1635 Enhancing the Quality of Silage Bales Produced by a Commercial Scale Silage Producer in Northern province, Sri Lanka: A Step Toward Supporting Smallholder Dairy Farmers in the Northern Province Sri Lanka

Authors: Harithas Aruchchunan

Abstract:

Silage production is an essential aspect of dairy farming, used to provide high-quality feed to ruminants. However, dairy farmers in Northern Province Sri Lanka are facing multiple challenges that compromise the quality and quantity of silage produced. To tackle these challenges, promoting silage feeding has become an essential component of sustainable dairy farming practices. In this study, silage bale samples were collected from a newly started silage baling factory in Jaffna, Northern province and their quality was analysed at the Veterinary Research Institute laboratory in Kandy in March 2023. The results show the nutritional composition of three Napier grass cultivars: Super Napier, CO6, and Indian Red Napier (BH18). The main parameters analysed were dry matter, pH, lactic acid, soluble carbohydrate, ammonia nitrogen, ash, crude protein, NDF, and ADF. The results indicate that Super Napier and CO6 have higher crude protein content and lower ADF levels, making them suitable for producing high-quality silage. The pH levels of all three cultivars were safe, and the ammonia nitrogen levels were considered appropriate. However, laboratory results indicate that the quality of silage bales produced can be further enhanced. Dairy farmers should be encouraged to adopt these cultivars to achieve better yields as they are high in protein and are better suited to Northern Province's soil and climate. Therefore, it is vital to educate small-scale fodder producers, who supply the raw material to silage factories, on the best practices of cultivating these new cultivars. To improve silage bale production and quality in Northern Province Sri Lanka, we recommend increasing public awareness about silage feeding, providing education and training to dairy farmers and small-scale fodder producers on modern silage production techniques and improving the availability of raw materials for silage production. Additionally, Napier grass cultivars need to be promoted among dairy farmers for better production and quality of silage bales. Failing to improve the quality and quantity of silage bale production could not only lead to the decline of dairy farming in Northern Province Sri Lanka but also the negative impact on the economy

Keywords: silage bales, dairy farming, economic crisis, Sri Lanka

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1634 The Response of Adaptive Mechanism of Fluorescent Proteins from Coral Species and Target Cell Properties on Signalling Capacity as Biosensor

Authors: Elif Tugce Aksun Tumerkan

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Fluorescent proteins (FPs) have become very popular since green fluorescent protein discovered from crystal jellyfish. It is known that Anthozoa species have a wide range of chromophore organisms, and the initial crystal structure for non-fluorescent chromophores obtained from the reef-building coral has been determined. There are also differently coloured pigments in non-bioluminescent Anthozoa zooxanthellate and azooxanthellate which are frequently members of the GFP-like protein family. The development of fluorescent proteins (FPs) and their applications is an outstanding example of basic science leading to practical biotechnological and medical applications. Fluorescent proteins have several applications in science and are used as important indicators in molecular biology and cell-based research. With rising interest in cell biology, FPs have used as biosensor indicators and probes in pharmacology and cell biology. Using fluorescent proteins in genetically encoded metabolite sensors has many advantages than chemical probes for metabolites such as easily introduced into any cell or organism in any sub-cellular localization and giving chance to fixing to fluoresce of different colours or characteristics. There are different factors effects to signalling mechanism when they used as a biosensor. While there are wide ranges of research have been done on the significance and applications of fluorescent proteins, the cell signalling response of FPs and target cell are less well understood. In this study, it was aimed to clarify the response of adaptive mechanisms of coral species such as pH, temperature and symbiotic relationship and target cells properties on the signalling capacity. Corals are a rich natural source of fluorescent proteins that change with environmental conditions such as light, heat stress and injury. Adaptation mechanism of coral species to these types of environmental variations is important factor due to FPs properties have affected by this mechanism. Since fluorescent proteins obtained from nature, their own ecological property like the symbiotic relationship is observed very commonly in coral species and living conditions have the impact on FPs efficiency. Target cell properties also have an effect on signalling and visualization. The dynamicity of detector that used for reading fluorescence and the level of background fluorescence are key parameters for the quality of the fluorescent signal. Among the factors, it can be concluded that coral species adaptive characteristics have the strongest effect on FPs signalling capacity.

Keywords: biosensor, cell biology, environmental conditions, fluorescent protein, sea anemone

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1633 Electron Density Discrepancy Analysis of Energy Metabolism Coenzymes

Authors: Alan Luo, Hunter N. B. Moseley

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Many macromolecular structure entries in the Protein Data Bank (PDB) have a range of regional (localized) quality issues, be it derived from x-ray crystallography, Nuclear Magnetic Resonance (NMR) spectroscopy, or other experimental approaches. However, most PDB entries are judged by global quality metrics like R-factor, R-free, and resolution for x-ray crystallography or backbone phi-psi distribution statistics and average restraint violations for NMR. Regional quality is often ignored when PDB entries are re-used for a variety of structurally based analyses. The binding of ligands, especially ligands involved in energy metabolism, is of particular interest in many structurally focused protein studies. Using a regional quality metric that provides chemically interpretable information from electron density maps, a significant number of outliers in regional structural quality was detected across x-ray crystallographic PDB entries for proteins bound to biochemically critical ligands. In this study, a series of analyses was performed to evaluate both specific and general potential factors that could promote these outliers. In particular, these potential factors were the minimum distance to a metal ion, the minimum distance to a crystal contact, and the isotropic atomic b-factor. To evaluate these potential factors, Fisher’s exact tests were performed, using regional quality criteria of outlier (top 1%, 2.5%, 5%, or 10%) versus non-outlier compared to a potential factor metric above versus below a certain outlier cutoff. The results revealed a consistent general effect from region-specific normalized b-factors but no specific effect from metal ion contact distances and only a very weak effect from crystal contact distance as compared to the b-factor results. These findings indicate that no single specific potential factor explains a majority of the outlier ligand-bound regions, implying that human error is likely as important as these other factors. Thus, all factors, including human error, should be considered when regions of low structural quality are detected. Also, the downstream re-use of protein structures for studying ligand-bound conformations should screen the regional quality of the binding sites. Doing so prevents misinterpretation due to the presence of structural uncertainty or flaws in regions of interest.

Keywords: biomacromolecular structure, coenzyme, electron density discrepancy analysis, x-ray crystallography

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1632 Establishment of Farmed Fish Welfare Biomarkers Using an Omics Approach

Authors: Pedro M. Rodrigues, Claudia Raposo, Denise Schrama, Marco Cerqueira

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Farmed fish welfare is a very recent concept, widely discussed among the scientific community. Consumers’ interest regarding farmed animal welfare standards has significantly increased in the last years posing a huge challenge to producers in order to maintain an equilibrium between good welfare principles and productivity, while simultaneously achieve public acceptance. The major bottleneck of standard aquaculture is to impair considerably fish welfare throughout the production cycle and with this, the quality of fish protein. Welfare assessment in farmed fish is undertaken through the evaluation of fish stress responses. Primary and secondary stress responses include release of cortisol and glucose and lactate to the blood stream, respectively, which are currently the most commonly used indicators of stress exposure. However, the reliability of these indicators is highly dubious, due to a high variability of fish responses to an acute stress and the adaptation of the animal to a repetitive chronic stress. Our objective is to use comparative proteomics to identify and validate a fingerprint of proteins that can present an more reliable alternative to the already established welfare indicators. In this way, the culture conditions will improve and there will be a higher perception of mechanisms and metabolic pathway involved in the produced organism’s welfare. Due to its high economical importance in Portuguese aquaculture Gilthead seabream will be the elected species for this study. Protein extracts from Gilthead Seabream fish muscle, liver and plasma, reared for a 3 month period under optimized culture conditions (control) and induced stress conditions (Handling, high densities, and Hipoxia) are collected and used to identify a putative fish welfare protein markers fingerprint using a proteomics approach. Three tanks per condition and 3 biological replicates per tank are used for each analisys. Briefly, proteins from target tissue/fluid are extracted using standard established protocols. Protein extracts are then separated using 2D-DIGE (Difference gel electrophoresis). Proteins differentially expressed between control and induced stress conditions will be identified by mass spectrometry (LC-Ms/Ms) using NCBInr (taxonomic level - Actinopterygii) databank and Mascot search engine. The statistical analysis is performed using the R software environment, having used a one-tailed Mann-Whitney U-test (p < 0.05) to assess which proteins were differentially expressed in a statistically significant way. Validation of these proteins will be done by comparison of the RT-qPCR (Quantitative reverse transcription polymerase chain reaction) expressed genes pattern with the proteomic profile. Cortisol, glucose, and lactate are also measured in order to confirm or refute the reliability of these indicators. The identified liver proteins under handling and high densities induced stress conditions are responsible and involved in several metabolic pathways like primary metabolism (i.e. glycolysis, gluconeogenesis), ammonia metabolism, cytoskeleton proteins, signalizing proteins, lipid transport. Validition of these proteins as well as identical analysis in muscle and plasma are underway. Proteomics is a promising high-throughput technique that can be successfully applied to identify putative welfare protein biomarkers in farmed fish.

Keywords: aquaculture, fish welfare, proteomics, welfare biomarkers

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1631 An Insight into the Interaction Study of a WhiB Protein and its Binding Partner

Authors: Sonam Kumari

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Tuberculosis is the deadliest disease worldwide. Millions of people lose their lives every year due to this disease. It has turned lethal due to the erratic nature of its causative organism, Mycobacterium tuberculosis (Mtb). Mtb tends to enter into an inactive, dormant state and emerge to replicating state upon encountering favorable conditions. The mechanism by which Mtb switches from the dormant state to the replicative form is still poorly characterized. Proteome studies have given us an insight into the role of certain proteins in giving stupendous virulence to Mtb, but numerous dotsremain unconnected and unaccounted. The WhiB family of proteins is one such protein that is associated with developmental processes in actinomycetes. Mtb has seven such proteins (WhiB1 to WhiB7). WhiB proteins are transcriptional regulators; they regulate various essential genes of Mtbby binding to their promoter DNA. Biophysical parameters of the effect of DNA binding on WhiB proteins has not yet been appropriately characterized. Interaction with DNA induces conformational changes in the WhiB proteins, confirmed by steady-state fluorescence and circular dichroism spectroscopy. ITC has deduced thermodynamic parameters and the binding affinity of the interaction. Since these transcription factors are highly unstable in vitro, their stability and solubility were enhanced by the co-expression of molecular chaperones. The present study findings help determine the conditions under which the WhiB proteins interact with their interacting partner and the factors that influence their binding affinity. This is crucial in understanding their role in regulating gene expression in Mtbandin targeting WhiB proteins as a drug target to cure TB.

Keywords: mycobacterium tuberculosis, TB, whiB proteins, ITC

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1630 The Omicron Variant BA.2.86.1 of SARS- 2 CoV-2 Demonstrates an Altered Interaction Network and Dynamic Features to Enhance the Interaction with the hACE2

Authors: Taimur Khan, Zakirullah, Muhammad Shahab

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The SARS-CoV-2 variant BA.2.86 (Omicron) has emerged with unique mutations that may increase its transmission and infectivity. This study investigates how these mutations alter the Omicron receptor-binding domain's interaction network and dynamic properties (RBD) compared to the wild-type virus, focusing on its binding affinity to the human ACE2 (hACE2) receptor. Protein-protein docking and all-atom molecular dynamics simulations were used to analyze structural and dynamic differences. Despite the structural similarity to the wild-type virus, the Omicron variant exhibits a distinct interaction network involving new residues that enhance its binding capacity. The dynamic analysis reveals increased flexibility in the RBD, particularly in loop regions crucial for hACE2 interaction. Mutations significantly alter the secondary structure, leading to greater flexibility and conformational adaptability compared to the wild type. Binding free energy calculations confirm that the Omicron RBD has a higher binding affinity (-70.47 kcal/mol) to hACE2 than the wild-type RBD (-61.38 kcal/mol). These results suggest that the altered interaction network and enhanced dynamics of the Omicron variant contribute to its increased infectivity, providing insights for the development of targeted therapeutics and vaccines.

Keywords: SARS-CoV-2, molecular dynamic simulation, receptor binding domain, vaccine

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1629 Efficiency of Natural Metabolites on Quality Milk Production in Mixed Breed Cows.

Authors: Mariam Azam, Sajjad Ur Rahman, Mukarram Bashir, Muhammad Tahir, Seemal Javaid, Jawad, Aoun Muhammad, Muhammad Zohaib, Hannan Khan

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Products of microbial origin are of great importance as they have proved their value in healthcare and nutrition, use of these microbial metabolites acquired from partially fermented soya hulls and wheat bran along with Saccharomyces cerevisiae (DL-22 S/N) substantiates to be a great source for an increase in the total milk production and quality yield.1×109 CFU/ml cells of Saccharomyces cerevisiae (DL-22 S/N) were further grown under in-vivo conditions for the assessment of quality milk production. Two groups with twelve cows, each having the same physical characteristics (Group A and Group B), were under study, Group A was daily fed with 12gm of biological metabolites and 22% protein-pelleted feed. On the other hand, the animals of Group B were provided with no metabolites in their feed. In thirty days of trial, improvement in the overall health, body score, milk protein, milk fat, yield, incidence rate of mastitis, ash, and solid not fat (SNF) was observed. The collected data showed that the average quality milk production was elevated up to 0.45 liter/h/d. However, a reduction in the milk fats up to 0.45% and uplift in the SNF value up to 0.53% of cow milk was also observed. At the same time, the incidence rate of mastitis recorded for the animals under trial was reduced to half, and improved non specific immunity was reported.

Keywords: microbial metabolites, post-biotics, animal supplements, animal nutrition, proteins, animal production, fermentation

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1628 Beneficial Effects of Whey Protein Concentrate in Venous Thrombosis

Authors: Anna Tokajuk, Agnieszka Zakrzeska, Ewa Chabielska, Halina Car

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Whey is a by-product generated mainly in the production of cheese and casein. Powder forms of whey are used widely in the food industry as well as a high-protein food for infants, for convalescents, by athletes and especially by bodybuilders to increase muscle mass during exercise. Whey protein concentrate-80 (WPC-80) is a source of bioactive peptides with beneficial effects on the cardiovascular system. It is known that whey proteins health beneficial properties include antidiabetic, blood pressure lowering, improving cardiovascular system function, antibacterial, antiviral and other effects. To study its influence on the development of thrombosis, venous thrombosis model was performed according to the protocol featured by Reyers with modification by Chabielska and Gromotowicz. Male Wistar-Crl: WI (Han) rats from researched groups were supplemented with two doses of WPC-80 (0.3 or 0.5 g/kg) for 7, 14 or 21 days and after these periods, one-hour venous thrombosis model was performed. Control group received 0.9 % NaCl solution and was sham operated. The statistical significance of results was computed by Mann – Whitney’s test. We observed that thrombus weight was decreased in animals obtaining WPC-8080 and that was statistically significant in 14 and 21-day supplemented groups. Blood count parameters did not differ significantly in rats with and without thrombosis induction whether they were fed with WPC-80 or not. Moreover, the number of platelets (PLT) was within the normal range in each group. The examined coagulation parameters in rats of the control groups were within normal limits. After WPC-80 supplementation there was the tendency to prolonged activated partial thromboplastin time (aPTT), but in comparison, the results did not turn out significant. In animals that received WPC-80 0.3 g·kg-1 for 21 days with and without induced thrombosis, prothrombin time (PT) and an international normalized ratio (INR) was somewhat decreased, remaining within the normal range, but the nature and significance of this observation are beyond the framework of the current study. Additionally, fibrinogen and thrombin time (TT) did not differ significantly between groups. Therefore the exact effect of WPC-80 on coagulation system is still elusive and requires further thorough research including mechanisms of action. Determining the potential clinical application of WPC-80 requires the selection of the optimal dose and duration of supplementation.

Keywords: antithrombotic, rats, venous thrombosis, WPC-80

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1627 Human Rabies Survivors in India: Epidemiological, Immunological and Virological Studies

Authors: Madhusudana S. N., Reeta Mani, Ashwini S. Satishchandra P., Netravati, Udhani V., Fiaz A., Karande S.

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Rabies is an acute encephalitis which is considered 100% fatal despite occasional reports of survivors. However, in recent times more cases of human rabies survivors are being reported. In the last 5 years, there are six laboratories confirmed human rabies survivors in India alone. All cases were children below 15 years and all contracted the disease by dog bites. All of them also had received the full or partial course of rabies vaccination and 4 out of 6 had also received rabies immunoglobulin. All cases were treated in intensive care units in hospitals at Bangalore, Mumbai, Chandigarh, Lucknow and Goa. We report here the results of immunological and virological studies conducted at our laboratory on these patients. The clinical samples that were obtained from these patients were Serum, CSF, nuchal skin biopsy and saliva. Serum and CSF samples were subjected to standard RFFIT for estimation of rabies neutralizing antibodies. Skin biopsy, CSF and saliva were processed by TaqMan real-time PCR for detection of viral RNA. CSF, saliva and skin homogenates were also processed for virus isolation by inoculation of suckling mice. The PBMCs isolated from fresh blood was subjected to ELISPOT assay to determine the type of immune response (Th1/Th2). Both CSF and serum were also investigated for selected cytokines by Luminex assay. The level of antibodies to virus G protein and N protein were determined by ELISA. All survivors had very high titers of RVNA in serum and CSF 100 fold higher than non-survivors and vaccine controls. A five-fold rise in titer could be demonstrated in 4 out of 6 patients. All survivors had a significant increase in antibodies to G protein in both CSF and serum when compared to non-survivors. There was a profound and robust Th1 response in all survivors indicating that interferon gamma could play an important factor in virus clearance. We could isolate viral RNA in only one patient four years after he had developed symptoms. The partial N gene sequencing revealed 99% homology to species I strain prevalent in India. Levels of selected cytokines in CSF and serum did not reveal any difference between survivors and non-survivors. To conclude, survival from rabies is mediated by virus-specific immune responses of the host and clearance of rabies virus from CNS may involve the participation of both Th2 and Th1 immune responses.

Keywords: rabies, rabies treatment, rabies survivors, immune reponse in rabies encephalitis

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1626 Effect of Different Phosphorus Levels on Vegetative Growth of Maize Variety

Authors: Tegene Nigussie

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Introduction: Maize is the most domesticated of all the field crops. Wild maize has not been found to date and there has been much speculation on its origin. Regardless of the validity of different theories, it is generally agreed that the center of origin of maize is Central America, primarily Mexico and the Caribbean. Maize in Africa is of a recent introduction although data suggest that it was present in Nigeria even before Columbus voyages. After being taken to Europe in 1493, maize was introduced to Africa and distributed (spread through the continent by different routes. Maize is an important cereal crop in Ethiopia in general, it is the primarily stable food, and rural households show strong preference. For human food, the important constituents of grain are carbohydrates (starch and sugars), protein, fat or oil (in the embryo) and minerals. About 75 percent of the kernel is starch, a range of 60.80 percent but low protein content (8-15%). In Ethiopia, the introduction of modern farming techniques appears to be a priority. However, the adoption of modern inputs by peasant farmers is found to be very slow, for example, the adoption rate of fertilizer, an input that is relatively adopted, is very slow. The difference in socio-economic factors lay behind the low rate of technological adoption, including price & marketing input. Objective: The aim of the study is to determine the optimum application rate or level of different phosphorus fertilizers for the vegetative growth of maize and to identify the effect of different phosphorus rates on the growth and development of maize. Methods: The vegetative parameter (above ground) measurement from five plants randomly sampled from the middle rows of each plot. Results: The interaction of nitrogen and maize variety showed a significant at (p<0.01) effect on plant height, with the application of 60kg/ha and BH140 maize variety in combination and root length with the application of 60kg/ha of nitrogen and BH140 variety of maize. The highest mean (12.33) of the number of leaves per plant and mean (7.1) of the number of nodes per plant can be used as an alternative for better vegetative growth of maize. Conclusion and Recommendation: Maize is one of the popular and cultivated crops in Ethiopia. This study was conducted to investigate the best dosage of phosphorus for vegetative growth, yield, and better quality of maize variety and to recommend a level of phosphorus rate and the best variety adaptable to the specific soil condition or area.

Keywords: leaf, carbohydrate protein, adoption, sugar

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1625 A pH-Activatable Nanoparticle Self-Assembly Triggered by 7-Amino Actinomycin D Demonstrating Superior Tumor Fluorescence Imaging and Anticancer Performance

Authors: Han Xiao

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The development of nanomedicines has recently achieved several breakthroughs in the field of cancer treatment; however, the biocompatibility and targeted burst release of these medications remain a limitation, which leads to serious side effects and significantly narrows the scope of their applications. The self-assembly of intermediate filament protein (IFP) peptides was triggered by a hydrophobic cation drug 7-amino actinomycin D (7-AAD) to synthesize pH-activatable nanoparticles (NPs) that could simultaneously locate tumors and produce antitumor effects. The designed IFP peptide included a target peptide (arginine–glycine–aspartate), a negatively charged region, and an α-helix sequence. It also possessed the ability to encapsulate 7-AAD molecules through the formation of hydrogen bonds and hydrophobic interactions by a one-step method. 7-AAD molecules with excellent near-infrared fluorescence properties could be target delivered into tumor cells by NPs and released immediately in the acidic environments of tumors and endosome/lysosomes, ultimately inducing cytotoxicity by arresting the tumor cell cycle with inserted DNA. It is noteworthy that the IFP/7-AAD NPs tail vein injection approach demonstrated not only high tumor-targeted imaging potential, but also strong antitumor therapeutic effects in vivo. The proposed strategy may be used in the delivery of cationic antitumor drugs for precise imaging and cancer therapy.

Keywords: 7-amino actinomycin D, intermediate filament protein, nanoparticle, tumor image

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1624 Improving Binding Selectivity in Molecularly Imprinted Polymers from Templates of Higher Biomolecular Weight: An Application in Cancer Targeting and Drug Delivery

Authors: Ben Otange, Wolfgang Parak, Florian Schulz, Michael Alexander Rubhausen

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The feasibility of extending the usage of molecular imprinting technique in complex biomolecules is demonstrated in this research. This technique is promising in diverse applications in areas such as drug delivery, diagnosis of diseases, catalysts, and impurities detection as well as treatment of various complications. While molecularly imprinted polymers MIP remain robust in the synthesis of molecules with remarkable binding sites that have high affinities to specific molecules of interest, extending the usage to complex biomolecules remains futile. This work reports on the successful synthesis of MIP from complex proteins: BSA, Transferrin, and MUC1. We show in this research that despite the heterogeneous binding sites and higher conformational flexibility of the chosen proteins, relying on their respective epitopes and motifs rather than the whole template produces highly sensitive and selective MIPs for specific molecular binding. Introduction: Proteins are vital in most biological processes, ranging from cell structure and structural integrity to complex functions such as transport and immunity in biological systems. Unlike other imprinting templates, proteins have heterogeneous binding sites in their complex long-chain structure, which makes their imprinting to be marred by challenges. In addressing this challenge, our attention is inclined toward the targeted delivery, which will use molecular imprinting on the particle surface so that these particles may recognize overexpressed proteins on the target cells. Our goal is thus to make surfaces of nanoparticles that specifically bind to the target cells. Results and Discussions: Using epitopes of BSA and MUC1 proteins and motifs with conserved receptors of transferrin as the respective templates for MIPs, significant improvement in the MIP sensitivity to the binding of complex protein templates was noted. Through the Fluorescence Correlation Spectroscopy FCS measurements on the size of protein corona after incubation of the synthesized nanoparticles with proteins, we noted a high affinity of MIPs to the binding of their respective complex proteins. In addition, quantitative analysis of hard corona using SDS-PAGE showed that only a specific protein was strongly bound on the respective MIPs when incubated with similar concentrations of the protein mixture. Conclusion: Our findings have shown that the merits of MIPs can be extended to complex molecules of higher biomolecular mass. As such, the unique merits of the technique, including high sensitivity and selectivity, relative ease of synthesis, production of materials with higher physical robustness, and higher stability, can be extended to more templates that were previously not suitable candidates despite their abundance and usage within the body.

Keywords: molecularly imprinted polymers, specific binding, drug delivery, high biomolecular mass-templates

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1623 Molecular Portraits: The Role of Posttranslational Modification in Cancer Metastasis

Authors: Navkiran Kaur, Apoorva Mathur, Abhishree Agarwal, Sakshi Gupta, Tuhin Rashmi

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Aim: Breast cancer is the most common cancer in women worldwide, and resistance to the current therapeutics, often concurrently, is an increasing clinical challenge. Glycosylation of proteins is one of the most important post-translational modifications. It is widely known that aberrant glycosylation has been implicated in many different diseases due to changes associated with biological function and protein folding. Alterations in cell surface glycosylation, can promote invasive behavior of tumor cells that ultimately lead to the progression of cancer. In breast cancer, there is an increasing evidence pertaining to the role of glycosylation in tumor formation and metastasis. In the present study, an attempt has been made to study the disease associated sialoglycoproteins in breast cancer by using bioinformatics tools. The sequence will be retrieved from UniProt database. A database in the form of a word document was made by a collection of FASTA sequences of breast cancer gene sequence. Glycosylation was studied using yinOyang tool on ExPASy and Differential genes expression and protein analysis was done in context of breast cancer metastasis. The number of residues predicted O-glc NAc threshold containing 50 aberrant glycosylation sites or more was detected and recorded for individual sequence. We found that the there is a significant change in the expression profiling of glycosylation patterns of various proteins associated with breast cancer. Differential aberrant glycosylated proteins in breast cancer cells with respect to non-neoplastic cells are an important factor for the overall progression and development of cancer.

Keywords: breast cancer, bioinformatics, cancer, metastasis, glycosylation

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1622 Evaluation of Systemic Immune-Inflammation Index in Obese Children

Authors: Mustafa M. Donma, Orkide Donma

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A growing list of cancers might be influenced by obesity. Obesity is associated with an increased risk for the occurrence and development of some cancers. Inflammation can lead to cancer. It is one of the characteristic features of cancer and plays a critical role in cancer development. C-reactive protein (CRP) is under evaluation related to the new and simple prognostic factors in patients with metastatic renal cell cancer. Obesity can predict and promote systemic inflammation in healthy adults. BMI is correlated with hs-CRP. In this study, SII index and CRP values were evaluated in children with normal BMI and those within the range of different obesity grades to detect the tendency towards cancer in pediatric obesity. A total of one hundred and ninety-four children; thirty-five children with normal BMI, twenty overweight (OW), forty-seven obese (OB) and ninety-two morbid obese (MO) participated in the study. Age- and sex-matched groups were constituted using BMI-for age percentiles. Informed consent was obtained. Ethical Committee approval was taken. Weight, height, waist circumference (C), hip C, head C and neck C of the children were measured. The complete blood count test was performed. C-reactive protein analysis was performed. Statistical analyses were performed using SPSS. The degree for statistical significance was p≤0.05. SII index values were progressively increasing starting from normal weight (NW) to MO children. There is a statistically significant difference between NW and OB as well as MO children. No significant difference was observed between NW and OW children, however, a correlation was observed between NW and OW children. MO constitutes the only group, which exhibited a statistically significant correlation between SII index and CRP. Obesity-related bladder, kidney, cervical, liver, colorectal, endometrial cancers are still being investigated. Obesity, characterized as a chronic low-grade inflammation, is a crucial risk factor for colon cancer. Elevated childhood BMI values may be indicative of processes leading to cancer, initiated early in life. Prevention of childhood adiposity may decrease the cancer incidence in adults. To authors’ best knowledge, this study is the first to introduce SII index values during obesity of varying degrees of severity. It is suggested that this index seems to affect all stages of obesity with an increasing tendency and may point out the concomitant status of obesity and cancer starting from very early periods of life.

Keywords: children, C-reactive protein, systemic immune-inflammation index, obesity

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1621 The Effect of Particulate Matter on Cardiomyocyte Apoptosis Through Mitochondrial Fission

Authors: Tsai-chun Lai, Szu-ju Fu, Tzu-lin Lee, Yuh-Lien Chen

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There is much evidence that exposure to fine particulate matter (PM) from air pollution increases the risk of cardiovascular morbidity and mortality. According to previous reports, PM in the air enters the respiratory tract, contacts the alveoli, and enters the blood circulation, leading to the progression of cardiovascular disease. PM pollution may also lead to cardiometabolic disturbances, increasing the risk of cardiovascular disease. The effects of PM on cardiac function and mitochondrial damage are currently unknown. We used mice and rat cardiomyocytes (H9c2) as animal and in vitro cell models, respectively, to simulate an air pollution environment using PM. These results indicate that the apoptosis-related factor PUMA, a regulator of apoptosis upregulated by p53, is increased in mice treated with PM. Apoptosis was aggravated in cardiomyocytes treated with PM, as measured by TUNEL assay and Annexin V/PI. Western blot results showed that CASPASE3 was significantly increased and BCL2 (B-cell lymphoid 2) was significantly decreased under PM treatment. Concurrent exposure to PM increases mitochondrial reactive oxygen species (ROS) production by MitoSOX Red staining. Furthermore, using Mitotracker staining, PM treatment significantly shortened mitochondrial length, indicating mitochondrial fission. The expression of mitochondrial fission-related proteins p-DRP1 (phosphodynamics-related protein 1) and FIS1 (mitochondrial fission 1 protein) was significantly increased. Based on these results, the exposure to PM worsens mitochondrial function and leads to cardiomyocyte apoptosis.

Keywords: particulate matter, cardiomyocyte, apoptosis, mitochondria

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1620 Influence of Surface Wettability on Imbibition Dynamics of Protein Solution in Microwells

Authors: Himani Sharma, Amit Agrawal

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Stability of the Cassie and Wenzel wetting states depends on intrinsic contact angle and geometric features on a surface that was exploited in capturing biofluids in microwells. However, the mechanism of imbibition of biofluids in the microwells is not well implied in terms of wettability of a substrate. In this work, we experimentally demonstrated filling dynamics in hydrophilic and hydrophobic microwells by protein solutions. Towards this, we utilized lotus leaf as a mold to fabricate microwells on a Polydimethylsiloxane (PDMS) surface. Lotus leaf containing micrometer-sized blunt-conical shaped pillars with a height of 8-15 µm and diameter of 3-8 µm were transferred on to PDMS. Furthermore, PDMS surface was treated with oxygen plasma to render the hydrophilic nature. A 10µL droplets containing fluorescein isothiocyanate (FITC) - labelled bovine serum albumin (BSA) were rested on both hydrophobic (θa = 108o, where θa is the apparent contact angle) and hydrophilic (θa = 60o) PDMS surfaces. A time-dependent fluorescence microscopy was conducted on these modified PDMS surfaces by recording the fluorescent intensity over a 5 minute period. It was observed that, initially (at t=1 min) FITC-BSA was accumulated on the periphery of both hydrophilic and hydrophobic microwells due to incomplete penetration of liquid-gas meniscus. This deposition of FITC-BSA on periphery of microwell was not changed with time for hydrophobic surfaces, whereas, a complete filling was occurred in hydrophilic microwells (at t=5 mins). This attributes to a gradual movement of three-phase contact line along the vertical surface of the hydrophilic microwells as compared to stable pinning in the hydrophobic microwells as confirmed by Surface Evolver simulations. In addition, if the cavities are presented on hydrophobic surfaces, air bubbles will be trapped inside the cavities once the aqueous solution is placed over these surfaces, resulting in the Cassie-Baxter wetting state. This condition hinders trapping of proteins inside the microwells. Thus, it is necessary to impart hydrophilicity to the microwell surfaces so as to induce the Wenzel state, such that, an entire solution will be fully in contact with the walls of microwells. Imbibition of microwells by protein solutions was analyzed in terms fluorescent intensity versus time. The present work underlines the importance of geometry of microwells and surface wettability of substrate in wetting and effective capturing of solid sub-phases in biofluids.

Keywords: BSA, microwells, surface evolver, wettability

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1619 Functional Profiling of a Circular RNA from the Huntingtin (HTT) Gene

Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

Abstract:

Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

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1618 Prevalence and Diagnostic Evaluation of Schistosomiasis in School-Going Children in Nelson Mandela Bay Municipality: Insights from Urinalysis and Point-of-Care Testing

Authors: Maryline Vere, Wilma ten Ham-Baloyi, Lucy Ochola, Opeoluwa Oyedele, Lindsey Beyleveld, Siphokazi Tili, Takafira Mduluza, Paula E. Melariri

Abstract:

Schistosomiasis, caused by Schistosoma (S.) haematobium and Schistosoma (S.) mansoni parasites poses a significant public health challenge in low-income regions. Diagnosis typically relies on identifying specific urine biomarkers such as haematuria, protein, and leukocytes for S. haematobium, while the Point-of-Care Circulating Cathodic Antigen (POC-CCA) assay is employed for detecting S. mansoni. Urinalysis and the POC-CCA assay are favoured for their rapid, non-invasive nature and cost-effectiveness. However, traditional diagnostic methods such as Kato-Katz and urine filtration lack sensitivity in low-transmission areas, which can lead to underreporting of cases and hinder effective disease control efforts. Therefore, in this study, urinalysis and the POC-CCA assay was utilised to diagnose schistosomiasis effectively among school-going children in Nelson Mandela Bay Municipality. This was a cross-sectional study with a total of 759 children, aged 5 to 14 years, who provided urine samples. Urinalysis was performed using urinary dipstick tests, which measure multiple parameters, including haematuria, protein, leukocytes, bilirubin, urobilinogen, ketones, pH, specific gravity and other biomarkers. Urinalysis was performed by dipping the strip into the urine sample and observing colour changes on specific reagent pads. The POC-CCA test was conducted by applying a drop of urine onto a cassette containing CCA-specific antibodies, and the presence of a visible test line indicated a positive result for S. mansoni infection. Descriptive statistics were used to summarize urine parameters, and Pearson correlation coefficients (r) were calculated to analyze associations among urine parameters using R software (version 4.3.1). Among the 759 children, the prevalence of S. haematobium using haematuria as a diagnostic marker was 33.6%. Additionally, leukocytes were detected in 21.3% of the samples, and protein was present in 15%. The prevalence of positive POC-CCA test results for S. mansoni was 3.7%. Urine parameters exhibited low to moderate associations, suggesting complex interrelationships. For instance, specific gravity and pH showed a negative correlation (r = -0.37), indicating that higher specific gravity was associated with lower pH. Weak correlations were observed between haematuria and pH (r = -0.10), bilirubin and ketones (r = 0.14), protein and bilirubin (r = 0.13), and urobilinogen and pH (r = 0.12). A mild positive correlation was found between leukocytes and blood (r = 0.23), reflecting some association between these inflammation markers. In conclusion, the study identified a significant prevalence of schistosomiasis among school-going children in Nelson Mandela Bay Municipality, with S. haematobium detected through haematuria and S. mansoni identified using the POC-CCA assay. The detection of leukocytes and protein in urine samples serves as critical biomarkers for schistosomiasis infections, reinforcing the presence of schistosomiasis in the study area when considered alongside haematuria. These urine parameters are indicative of inflammatory responses associated with schistosomiasis, underscoring the necessity for effective diagnostic methodologies. Such findings highlight the importance of comprehensive diagnostic assessments to accurately identify and monitor schistosomiasis prevalence and its associated health impacts. The significant burden of schistosomiasis in this population highlights the urgent need to develop targeted control interventions to effectively reduce its prevalence in the study area.

Keywords: schistosomiasis, urinalysis, haematuria, POC-CCA

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1617 Oleuropein Ameliorates Palmitate-Induced Insulin Resistance by Increasing GLUT4 Translocation through Activation of AMP-Activated Protein Kinase in Rat Soleus Muscles

Authors: Hakam Alkhateeb

Abstract:

Oleuropein, the main constituent of leaves and fruits of the olive tree, has been demonstrated to exert beneficial effects on parameters relevant to the normal homeostatic mechanisms of glucose regulation in rat skeletal muscle. However, the antidiabetic effect of oleuropein, to our knowledge, has not been examined. Therefore, in this study, we examined whether oleuropein ameliorated palmitate-induced insulin resistance in skeletal muscle. To examine this question, insulin resistance was rapidly induced by incubating (12h) soleus muscle with a high concentration of palmitate(2mM). Subsequently, we attempted to restore insulin sensitivity by incubating (12h) muscles with oleuropien (1.5mM), while maintaining high concentrations of palmitate. Palmitate treatment for 12 h reduced insulin-stimulated glucose transport, GLUT4 translocationandAS160 phosphorylation. Oleuropein treatment (12 h) fully restoredinsulin-stimulated glucose transport, GLUT4translocationandAS160 phosphorylation. Inhibition of PI3K phosphorylation with wortmannin (1µM)did not affect the oleuropein-induced improvements in insulin-stimulated glucose transport, GLUT4 translocation, and AS160 phosphorylation. These results suggested that the improvements in these parameters cannot account for activating PI3K pathway. Taken altogether, it appears that oleuropein, through activation of another pathway like activated protein kinase (AMPK), may provide a possible strategy by which they ameliorate palmitate-induced insulin resistance in skeletal muscles.

Keywords: AS160, diabetes, GLUT4, oleuropein

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1616 Docking, Pharmacophore Modeling and 3d QSAR Studies on Some Novel HDAC Inhibitors with Heterocyclic Linker

Authors: Harish Rajak, Preeti Patel

Abstract:

The application of histone deacetylase inhibitors is a well-known strategy in prevention of cancer which shows acceptable preclinical antitumor activity due to its ability of growth inhibition and apoptosis induction of cancer cell. Molecular docking were performed using Histone Deacetylase protein (PDB ID:1t69) and prepared series of hydroxamic acid based HDACIs. On the basis of docking study, it was predicted that compound 1 has significant binding interaction with HDAC protein and three hydrogen bond interactions takes place, which are essential for antitumor activity. On docking, most of the compounds exhibited better glide score values between -8 to -10 which is close to the glide score value of suberoylanilide hydroxamic acid. The pharmacophore hypotheses were developed using e-pharmacophore script and phase module. The 3D-QSAR models provided a good correlation between predicted and actual anticancer activity. Best QSAR model showed Q2 (0.7974), R2 (0.9200) and standard deviation (0.2308). QSAR visualization maps suggest that hydrogen bond acceptor groups at carbonyl group of cap region and hydrophobic groups at ortho, meta, para position of R9 were favorable for HDAC inhibitory activity. We established structure activity correlation using docking, pharmacophore modeling and atom based 3D QSAR model for hydroxamic acid based HDACIs.

Keywords: HDACIs, QSAR, e-pharmacophore, docking, suberoylanilide hydroxamic acid

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1615 Role of Moderate Intensity Exercises in the Amelioration of Oxidant-Antioxidant Status and the Levels of Inflammatory Cytokines in Rheumatoid Arthritis Patients

Authors: Somaiya Mateen, Shagufta Moin, Abdul Qayyum, Atif Zafar

Abstract:

Cytokines and reactive species play an important role in the pathophysiology of rheumatoid arthritis (RA). This study was done to determine the levels of reactive oxygen and nitrogen species (ROS and RNS), inflammatory cytokines and the markers of protein, DNA and lipid oxidation in the blood of RA patients, with the aim to study the antioxidant and anti-inflammatory role of moderate intensity exercises in the management of RA. RA patients were subdivided into two groups- first group (n=30) received treatment with conventional RA drugs while the second group (n=30) received moderate exercise therapy along with the conventional drugs for a period of 12 weeks. The levels of ROS, RNS, inflammatory cytokines and markers of biomolecule oxidation were monitored before and after 12 weeks of treatment. RA patients showed a marked increase in the levels of ROS, RNS, inflammatory cytokines, lipid, protein and DNA oxidation as compared to the healthy controls. These parameters were ameliorated after treatment with drugs alone and exercise combined with drugs, with the amelioration being more significant in patients given drugs along with the moderate intensity exercise treatment. In conclusion, the role of ROS, RNS and inflammatory cytokines in the pathogenesis of RA has been confirmed by this study. These may also serve as potential biomarker for assessing the disease severity. Finally, the addition of moderate intensity exercises in the management of RA may be of great value.

Keywords: rheumatoid arthritis, reactive oxygen species, inflammatory cytokines, moderate intensity exercises

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1614 Artificial Cells Capable of Communication by Using Polymer Hydrogel

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

Abstract:

The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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1613 Field Effects on Seed Germination of Phaseolus Vulgaris, Early Seedling Growth and Chemical Composition

Authors: Najafi S., Heidai R., Jamei R., Tofigh F.

Abstract:

In order to study the effects of magnetic field on the root system and growth of Phaseolus vulgaris, an experiment was conducted in 2012. The possible involvement of magnetic field (MF) pretreatment in physiological factors of Phaseolus vulgaris was investigated. Seeds were subjected to 10 days with 1.8 mT of magnetic field for 1h per day. MF pretreatment decreased the plant height, fresh and dry weight, length of root and length of shoot, Chlorophyll a, Chlorophyll b and carotenoid in 10 days old seedling. In addition, activity of enzymes such as Catalase and Guaiacol peroxidase was decreased due to MF exposure. Also, the total Protein and DPPH content of the treated by magnetic field was not significantly changed in compare to control groups, while the flavonoid, Phenol and prolin content of the treated of the treated by magnetic field was significantly changed in compare to control groups. Lateral branches of roots and secondary roots increased with MF. The results suggest that pretreatment of this MF plays important roles in changes in crop productivity. In all cases there was observed a slight stimulating effect of the factors examined. The growth dynamics were weakened. The plants were shorter. Moreover, the effect of a magnetic field on the crop of Phaseolus vulgaris and its structure was small.

Keywords: carotenoid, Chlorophyll a, Chlorophyll b, DPPH, enzymes, flavonoid, germination, growth, phenol, proline, protein, magnetic field, phaseolus vulgaris

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1612 Fermented Unripe Plantain (Musa paradisiacal) Peel Meal as a Replacement for Maize in the Diet of Nile Tilapia (Oreochromis niloticus) Fingerlings

Authors: N. A. Bamidele, S. O. Obasa, I. O. Taiwo, I. Abdulraheem, O. C. Odebiyi, A. A. Adeoye, O. E. Babalola, O. V. Uzamere

Abstract:

A feeding trial was conducted to investigate the effect of fermented unripe plantain peel meal (FUP) on growth performance, nutrients digestibility and economic indices of production of Nile tilapia, Oreochromis niloticus fingerlings. Fingerlings (150) of Nile tilapia (1.70±0.1g) were stocked at 10 per plastic tank. Five iso-nitrogenous diets containing 40% crude protein in which maize meal was replaced by fermented unripe plantain peel meal at 0% (FUP0), 25% (FUP25), 50% (FUP50), 75% (FUP75) and 100% (FUP100) were formulated and prepared. The fingerlings were fed at 5% body weight per day for 56 days. There was no significant difference (p > 0.05) in all the growth parameters among the treatments. Feed conversion ratio of 1.35 in fish fed diet FUP25 was not significantly different (P > 0.05) from 1.42 of fish fed diet FUP0. Apparent protein digestibility of 86.94% in fish fed diet FUP100 was significantly higher (p < 0.05) than 70.37% in fish fed diet FUP0 while apparent carbohydrate of 88.34% in fish fed diet FUP0 was significantly different (p < 0.05) from 70.29% of FUP100. Red blood cell (4.30 ml/mm3) of fish fed diet FUP100 was not significantly different from 4.13 ml/mm3 of fish fed diet FUP50. The highest percentage profit of 88.85% in fish fed diet FUP100 was significantly higher than 66.68% in fish fed diet FUP0 while the profit index of 1.89 in fish fed diet FUP100 was significantly different from 1.67 in fish fed diet FUP0. Therefore, fermented unripe plantain peel meal can completely replace maize in the diet of O. niloticus fingerlings.

Keywords: fermentation, fish diets, plantain peel, tilapia

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1611 Pregnancy Nutritional Status in Ethiopia: A Case Study of Pregnant Women in Shashemene District, Southern Oromia Region

Authors: Yoseph Gela Ali

Abstract:

Inadequate quality and quantity diet is one of the major reasons for high levels of malnutrition in pregnant women. Across-sectional survey was conducted in Shashemene District, Southern Oromia Region. A two-stage cluster sampling technique was used to select a representative sample of 15190 pregnant women aged 20-50 years from four rural villages Energy and nutrient intakes from foods were calculated from one-day weighed food records on a sub-sample (n = 83). The result of the study showed that the intakes of most nutrients were lower than the recommended intake. The energy intake of the study participants both in 2nd and 3rd trimesters of pregnancy were 2,308 kcal and 1,420.5 kcal compared to the recommended 2,340 kcal and 2,452 kcal, respectively. Except iron, almost all micronutrient intakes were lower than the recommended intake. Vitamin A intake was 3 µg compared with the recommended 800 µg, while protein intake of the study respondents in 2nd and 3rd trimester of pregnancy was 45.9 g and 31.5 g, respectively, compared with the recommended 71 g. Risk factors for under nutrition were multiple pregnancy and no consumption of cereal-based foods. This study revealed that the energy and nutrient intake of the pregnant women in the study area was below the recommended intakes. Furthermore, the situation might be aggravated by the high phytate content food consumption reported. Nutritional status of pregnant women in the study area was not adequate to support the increased energy and nutrient requirement of the participants.

Keywords: nutrition, pregnancy, protein, vitamin, energy

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1610 Effects of Exercise Training in the Cold on Browning of White Fat in Obese Rats

Authors: Xiquan Weng, Chaoge Wang, Guoqin Xu, Wentao Lin

Abstract:

Objective: Cold exposure and exercise serve as two powerful physiological stimuli to launch the conversion of fat-accumulating white adipose tissue (WAT) into energy-dissipating brown adipose tissue (BAT). So far, it remains to be elucidated whether exercise plus cold exposure can produce an addictive effect on promoting WAT browning. Methods: 64 SD rats were subjected to high-fat and high-sugar diets for 9-week and successfully established an obesity model. They were randomly divided into 8 groups: normal control group (NC), normal exercise group (NE), continuous cold control group (CC), continuous cold exercise group (CE), intermittent cold control group (IC) and intermittent cold exercise group (IE). For continuous cold exposure, the rats stayed in a cold environment all day; For intermittent cold exposure, the rats were exposed to cold for only 4h per day. The protocol for treadmill exercises were as follows: 25m/min (speed), 0°C (slope), 30mins each time, an interval for 10 mins between two exercises, twice/two days, lasting for 5 weeks. Sampling were conducted on the 5th weekend. The body length and weight of the rats were measured, and the Lee's index was calculated. The visceral fat rate (VFR), subcutaneous fat rate (SFR), brown fat rate (BrFR) and body fat rate (BoFR) were measured by Micro-CT LCT200, and the expression of UCP1 protein in inguinal fat was examined by Western-blot. SPSS 22.0 was used for statistical analysis of the experimental results, and the ANOVA analysis was performed between groups (P < 0.05 was significant). Results: (1) Compared with the NC group, the weight of obese rats was significantly declined in the NE, CE and IE groups (P < 0.05), the Lee's index of obese rats significantly declined in the CE group (P < 0.05). Compared with the NE group, the weight of obese rats was significantly declined in the CE and IE groups (P < 0.05). (2)Compared with the NC group, the VFR and BoFR of the rats significantly declined in the NE, CE and IE groups (P < 0.05), the SFR of the rats significantly declined in the CE and IE groups (P < 0.05), and the BFR of the rats was significantly higher in the CC and IC groups (P < 0.05), respectively. Compared with the NE group, the VFR and BoFR of the rats significantly declined in the CE group (P < 0.05), the SFR of the rats was significantly higher in the CC and IS groups (P < 0.05), and the BrFR of the rats was significantly higher in the IC group (P < 0.05). (3)Compared with the NC group, the up-regulation of UCP1 protein expression in the inguinal fat of the rats was significant in the NE, CC, CE, IC and IE groups (P < 0.05). Compared with the NE group, the up-regulation of UCP1 protein expression in the inguinal fat of the rats was significant in the CC, CE and IE groups (P < 0.05). Conclusions: Exercise in the continuous and intermittent cold, especially in the former, can effectively decline the weight and body fat rate of obese rats. This is related to the effect of cold and exercise on the browning of white fat in rats.

Keywords: cold, browning of white fat, exercise, obesity

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1609 Effects of Cymbopogon citratus, Stapf (CS) or Lemon Grass Ethanol Extract on Antioxidant and Vascular Disorders Parameters in Rat

Authors: Suphaket Saenthaweesuk, Nutiya Somparn, Atcharaporn Thewmore

Abstract:

The present study aims to investigate the effects of Cymbopogon citratus, Stapf (CS) or lemon grass ethanol extract on antioxidant and vascular disorders parameters in rat. The CS ethanol extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with CS at 1,000 mg/kg/day for 30 days. Phytochemical screening of CS extract indicated the presence of tannins, flavonoids and phenolic compounds. The extract contained phenolic compounds 1,400.10 ± 0.47 mg of gallic acid equivalents per gram CS extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 168.77 ± 3.32µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14 µg/mL. In the animals, the protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased. This was consistent with elevation of serum catalase (CAT) and superoxide dismutase (SOD) activities. However, Protein expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and endothelial nitric oxide synthase (eNOS) in heart and aorta were not differenced from normal control. Taken together, the present study provides evidence that CCS water extract exhibits direct antioxidant properties and can induce cytoprotective enzymes in vivo.

Keywords: antioxidant, Cymbopogon citratus Stapf, VCAM-1, γ-glutamylcysteine ligase

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1608 Nutritive Value of Three-Stage Olive Cake (Olea europaea L.) for Growing Rabbit

Authors: Zahia Dorbane, Si Ammar Kadi, Dalila Boudouma, Thierry Gidenne

Abstract:

In rabbits feeding, minimum fibre intake is essential to avoid digestive disorders. However, this concentration of fibre is not easy to obtain when formulating feeds, without reduction of nutritional value. Three stage olive cake, the residual material after oil extraction by centrifugation, including pulp and stones, can be used as a fibre source in rabbit diet. The incorporation of olive cake can allow a better balance between different fibre fractions and reduce health disorder. However, for practical use of any raw material, it is necessary to know its chemical and nutritive value. The aim of this study was to assess the nutritive value of three-stage olive cake (TSOC) for growing rabbits. Thus, 36 rabbits weaned at 35 days (702.8 ± 28.5) were divided into three groups of 12 receiving one of the following diets: control with 0% of TSOC, TSOC10 (10% of TSOC) and TSOC20 (20% TSOC). The rabbits were individually housed in digestibility cages and received ad libitum one of the three diets, fresh and clean water was provided ad libitum. After an adaptation period of 7d, feces were collected for 4d. Collected feces were frozen and stored for further analysis. The chemical composition of TSOC shows that it is a rich fiber raw material since it contains (%DM): 6% of CP; 7.4% of EE; 78.7% of NDF; 55.4% of ADF and 24.3% of ADL. The inclusion of TSOC at 20% of basal diet reduced the digestibility coefficient of organic matter, crude protein and NDF from 67.8 to 55.3%, 80.4 to 75.3% and from 31.5 to 18.4% (p < 0.001) respectively. The digestible energy and digestible protein content of the three-stage olive cake estimated by regression was 2.94 ± 0.52MJ DE/kg DM and 22.4 ± 6 g DP/kg DM respectively. In conclusion, based on the results of the present experiment, the three-stage olive cake can be used as a fibre source for rabbit.

Keywords: digestibility, nutritive value, olive cake, rabbit

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1607 Effect of Phosphorus and Potassium Nutrition on Growth, Yield and Minerals Accumulation of Two Soybean Cultivars Differing in Phytate Contents

Authors: Taliman Nisar Ahmad, Hirofume Saneoka

Abstract:

A pot experiment was conducted to investigate the effect of phosphorus (P) and potassium (K) nutrition on grain yield, phytic acid and grain quality of high-phytate (Akimaro) and low-phytate line. Phosphorus and potassium were applied as; P₁ (20 kg ha⁻¹) and P₂ (100 kg ha⁻¹), same as K₁ (20 kg ha⁻¹) and K₂ (100 kg ha⁻¹), respectively. Low-phytate soybean had the highest grain yield, and 75% increase was observed compared to the high-phytate under same treatments. Highly significant differences of seed phytate P were observed in both cultivars, and the phytate P in high-phytate was found 39% higher than low-phytate, whereas no significant differences observed in response to P and K treatment. Percentage of phytate P from total P in seeds was 28 to 35% in low-phytate and 72 to 81% in high-phytate in different treatments. The lipid content in low-phytate was found lowered compared to that of high-phytate. Crude protein in grains was also found significantly higher in PK combined. No significant difference was observed in seed calcium (Ca), magnesium (Mg), and Zinc (Zn) in different treatments, but high-phytate showed 87% increase in seed Ca and 76% of Mg compared to low-phytate; however, low-phytate showed 82% increase in Zn content over high-phytate. The result illustrates that low-phytate soybean achieved higher grain yield and grain Pi in response to increased P and K nutrition. To achieve higher yield and quality seeds from the low-phytate soybean, it is recommended that proper phosphorus and potassium nutrition to be applied suggested in this study.

Keywords: phytic acid, low-phytate soybean, high-phytate soybean, P and K nutrition, protein content, soybean

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1606 Review of the Nutritional Value of Spirulina as a Potential Replacement of Fishmeal in Aquafeed

Authors: Onada Olawale Ahmed

Abstract:

As the intensification of aquaculture production increases on global scale, the growing concern of fish farmers around the world is related to cost of fish production, where cost of feeding takes substantial percentage. Fishmeal (FM) is one of the most expensive ingredients, and its high dependence in aqua-feed production translates to high cost of feeding of stocked fish. However, to reach a sustainable aquaculture, new alternative protein sources including cheaper plant or animal origin proteins are needed to be introduced for stable aqua-feed production. Spirulina is a cyanobacterium that has good nutrient profile that could be useful in aquaculture. This review therefore emphasizes on the nutritional value of Spirulina as a potential replacement of FM in aqua-feed. Spirulina is a planktonic photosynthetic filamentous cyanobacterium that forms massive populations in tropical and subtropical bodies of water with high levels of carbonate and bicarbonate. Spirulina grows naturally in nutrient rich alkaline lake with water salinity ( > 30 g/l) and high pH (8.5–11.0). Its artificial production requires luminosity (photo-period 12/12, 4 luxes), temperature (30 °C), inoculum, water stirring device, dissolved solids (10–60 g/litre), pH (8.5– 10.5), good water quality, and macro and micronutrient presence (C, N, P, K, S, Mg, Na, Cl, Ca and Fe, Zn, Cu, Ni, Co, Se). Spirulina has also been reported to grow on agro-industrial waste such as sugar mill waste effluent, poultry industry waste, fertilizer factory waste, and urban waste and organic matter. Chemical composition of Spirulina indicates that it has high nutritional value due to its content of 55-70% protein, 14-19% soluble carbohydrate, high amount of polyunsaturated fatty acids (PUFAs), 1.5–2.0 percent of 5–6 percent total lipid, all the essential minerals are available in spirulina which contributes about 7 percent (average range 2.76–3.00 percent of total weight) under laboratory conditions, β-carotene, B-group vitamin, vitamin E, iron, potassium and chlorophyll are also available in spirulina. Spirulina protein has a balanced composition of amino acids with concentration of methionine, tryptophan and other amino acids almost similar to those of casein, although, this depends upon the culture media used. Positive effects of spirulina on growth, feed utilization and stress and disease resistance of cultured fish have been reported in earlier studies. Spirulina was reported to replace up to 40% of fishmeal protein in tilapia (Oreochromis mossambicus) diet and even higher replacement of fishmeal was possible in common carp (Cyprinus carpio), partial replacement of fish meal with spirulina in diets for parrot fish (Oplegnathus fasciatus) and Tilapia (Orechromis niloticus) has also been conducted. Spirulina have considerable potential for development, especially as a small-scale crop for nutritional enhancement and health improvement of fish. It is important therefore that more research needs to be conducted on its production, inclusion level in aqua-feed and its possible potential use of aquaculture.

Keywords: aquaculture, spirulina, fish nutrition, fish feed

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