Search results for: protein kinase C alpha

Authors: Samantha A. Cintron, Janet Pierce, Mihaela E. Sardiu, Diane Mahoney, Jill Peltzer, Bhanu Gupta, Qiuhua Shen

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High output heart failure (HOHF) is characterized a high output state resulting from an underlying disease process and is commonly caused by obesity. As obesity levels increase, more individuals will be at risk for obesity-related HOHF. However, the underlying pathophysiologic mechanisms of obesity-related HOHF are not well understood and need further research. The aim of the study was to describe the differences in leukocyte transcriptomes of morbidly obese patients with HOHF and those with non-HOHF. In this cross-sectional study, the study team collected blood samples, demographics, and clinical data of six patients with morbid obesity and HOHF and six patients with morbid obesity and non-HOHF. The study team isolated the peripheral blood leukocyte RNA and applied stranded total RNA sequencing. Differential gene expression was calculated, and Ingenuity Pathway Analysis software was used to interpret the canonical pathways, functional changes, upstream regulators, and mechanistic and causal networks that were associated with the significantly different leukocyte transcriptomes. The study team identified 116 differentially expressed genes; 114 were upregulated, and 2 were downregulated in the HOHF group (Benjamini-Hochberg adjusted p-value ≤ 0.05 and log2(fold-change) of ±1). The differentially expressed genes were involved with cell proliferation, mitochondrial function, erythropoiesis, erythrocyte stability, and apoptosis. The top upregulated canonical pathways associated with differentially expressed genes were autophagy, adenosine monophosphate-activated protein kinase signaling, and senescence pathways. Upstream regulator GATA Binding Protein 1 (GATA1) and a network associated with nuclear factor kappa-light chain-enhancer of activated B cells (NF-kB) were also identified based on the different leukocyte transcriptomes of morbidly obese patients with HOHF and non-HOHF. To the author’s best knowledge, this is the first study that reported the differential gene expression in patients with obesity-related HOHF and demonstrated the unique pathophysiologic mechanisms underlying the disease. Further research is needed to determine the role of cellular function and maintenance, inflammation, and iron homeostasis in obesity-related HOHF.

Keywords: cardiac output, heart failure, obesity, transcriptomics

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Authors: Muhammad A. Khan, Waseem Shahzad

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Analysis of protein sequences is carried out for the purpose to discover their structural and ancestry relationship. Sequence similarity determines similar protein structures, similar function, and homology detection. Biological sequences composed of amino acid residues or nucleotides provide significant information through sequence alignment. In this paper, we present a new similarity/dissimilarity measure to sequence alignment based on the primary structure of a protein. The approach finds the distance between the two given sequences using the novel sequence alignment algorithm and a mathematical model. The algorithm runs at a time complexity of O(n²). A distance matrix is generated to construct a phylogenetic tree of different species. The new similarity/dissimilarity measure outperforms other existing methods.

Keywords: alignment, distance, homology, mathematical model, phylogenetic tree

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Authors: A. Zurraini, Syed Alwi Sar, H. Helmy, H. Nazeefah

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Early Dementia Questionnaire (EDQ) was developed as a screening tool to detect patients with early dementia in primary care. It was developed based on 20 symptoms of dementia. From a preliminary study, EDQ had been shown to be a promising alternative for screening of early dementia. This study was done to further test on EDQ’s reliability and validity. Using a systematic random sampling, 200 elderly patients attending primary health care centers in Kuching, Sarawak had consented to participate in the study and were administered the EDQ. Geriatric Depression Scale (GDS) was used to exclude patients with depression. Those who scored >21 MMSE, were retested using the EDQ. Reliability was determined by Cronbach’s alpha for internal consistency and construct validity was assessed using confirmatory factor analysis (principle component with varimax rotation). The result showed that the overall Cronbach’s alpha coefficient was good which was 0.874. Confirmatory factor analysis on 4 factors indicated that the Cronbach’s alpha for each domain were acceptable with memory (0.741), concentration (0.764), emotional and physical symptoms (0.754) and lastly sleep and environment (0.720). Pearson correlation coefficient between the first EDQ score and the retest EDQ score among those with MMSE of >21 showed a very strong, positive correlation between the two variables, r = 0.992, N=160, P <0.001. The results of the validation study showed that Early Dementia Questionnaire (EDQ) is a valid and reliable tool to be used as a screening tool to detect early dementia in primary care.

Keywords: Early Dementia Questionnaire (EDQ), screening, primary care, construct validity

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Authors: Tariq Ali Gujar, Anita Hökelmann

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Postural balance plays essential role throughout life in daily activities. Somatosensory, visual and vestibular inputs play the fundamental role in maintaining body equilibrium to balance the posture. The aim of this study was to find out electroencephalography (EEG) responses during balance activity of young people during Sensory Organization Balance Test. The outcome of this study will help to create the fitness and neurorehabilitation plan. 25 young people (25 ± 3.1 years) have been analyzed on Balance Master NeuroCom® with the coupling of Brain Vision 32 electrode wireless EEG system during the Sensory Organization Test. From the results it has been found that the balance score of samples is significantly higher under the influence of somatosensory input as compared to visual and vestibular input (p < 0.05). The EEG between somatosensory and visual input to balance the posture showed significantly higher (p < 0.05) alpha and beta activities during somatosensory input in somatosensory, attention and visual functions of the cortex whereas executive and motor functions of the cerebral cortex showed significantly higher (p < 0.05) alpha EEG activity during the visual input. The results suggest that somatosensory and attention function of the cerebral cortex has alpha and beta activity, respectively high during somatosensory and vestibular input in maintaining balance. In patients with balance impairments both physical and cognitive training, including neurofeedback will be helpful to improve balance abilities.

Keywords: balance, electroencephalography activity, somatosensory, visual, vestibular

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Authors: Dicky Eka Putra, S. P. Nadia Chairunissa, Lidia Paramita, Roza Hartati, Ice Yolanda Puri

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Background: Soy bean and the products such as tofu, tempeh and soy milk are famous in the community. Moreover, another product is tofu flour which is not familiar in Indonesia yet and it is well known as Okara. There are massive differences of energy, protein and carbohydrate between them which is know as good for protein sources as well. Unfortunately, it is seldom used as food variety. Basically, it can be benefit in order to create many products for example cakes, snacks and some desserts. Aim: the study was in order to promote the benefit of tofu flour as school feeding of elementary school and baby porridge and also to compare the nutrient. Method: Soy pulp was filtered and steamed approximately 30 minutes. Then, it was put at a plate under sunrise or barked on the oven for 10 hours at 800C. When it have dried and milling and tofu flour is ready to be used. Result: Tofu flour could be used as substitute of flour and rice flour when people want to cook some foods. In addition, some references said that soy bean is good for a specific remedy for the proper functioning of the heart, liver, kidneys, stomach, and bowels, constipation, as a stimulant for the lungs, for eradication of poison from the system, improving the complexion by cleaning the skin of impurities, and stimulating the growth and appearance of the hair. Discussion: Comparing between soy bean, tofu and tofu flour which has difference amount of nutrients. For example energy 382 kcal, 79 kcal and 393 kcal respectively and also protein 30.2 kcal, 7.8 kcal, and 17.4 kcal. In addition, carbohydrate of soy pulp was high than soy bean and tofu (30.1 kcal). Finally, local should replace flour, rice and gelatin rice flour with tofu flour.

Keywords: tofu flour, protein, soy bean, school feeding

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Authors: Meziani Samira, Menadi Noreddine, Labga Lahouaria, Chenni Fatima Zohra, Toumi Asma

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A comparative study between two varieties of soya beans was carried out in this work. The method consists of studying and proceeding to prepare a by-product (Flour) from two varieties of soybeans, a Chinese variety imported and marketed in Algeria. The chemical composition of ash, protein and fat was determined in this study. The minerals, namely potassium and sodium, were measured by flame spectrophotometer. In addition, the estimation of the polyphenol content and evaluation of the antioxidant activity Ferric Reducing Antioxidant Power assay (FRAP) f the methanol extracts of the flours were also carried out. The result revealed that soy flour from two cultivars, on average, contained 8% moisture, more than 50% protein, 1.58-1.87g fat, and 0.28-0.30g of ash. A slight difference was found for contents of 489 mg/ml of K + and 20 mg/ml of NA +. In addition, the phenolic content of the methanolic extracts gives a value of almost 37 mg EAG / g for both cultivars of soy flour. The estimated Reductive Antioxidant Iron (FRAP) potency of soy flour might be related to its polyphenol richness, which is similar to the variety of China. The flour Soya varieties tested contained a significant amount of protein and phenolic compounds with good antioxidant properties.

Keywords: soye beans, soya flour, protein, total polyphenols

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Authors: Alyaa Abdlwahab

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Cutaneous leishmaniasis represents a serious health problem in Syria; this problem has become noticeably aggravated after the civil war in the country. Leishmania tropica parasite is the main cause of cutaneous leishmaniasis in Syria. In order to control the disease, we need an effective vaccine against leishmania parasite. DNA vaccination remains one of the favorable approaches that have been used to face cutaneous leishmaniasis. Ribosomal protein S4 is responsible for important roles in Leishmania parasite life. DNA vaccine based on S4 gene has been used against infections by many species of Leishmania parasite but leishmania tropica parasite, so this gene represents a good candidate for DNA vaccine construction. After proving the existence of ribosomal protein S4 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), sequencing it and cloning it into pCI plasmid, BALB/C mice were inoculated with pCI-S4 DNA vaccine. The immune response was determined by monitoring the lesion progression in inoculated BALB/C mice for six weeks after challenging mice with Leishmania tropica (LCED Syrian 01) parasites. IL-12, IFN-γ, and IL-4 were quantified in draining lymph nodes (DLNa) of the immunized BALB/C mice by using the RT-qPCR technique. The parasite burden was calculated in the final week for the footpad lesion and the DLNs of the mice. This study proved the existence and the expression of the ribosomal protein S4 gene in Leishmania tropica (LCED Syrian 01) promastigotes. The sequence of ribosomal protein cDNA S4 gene was determined and published in Genbank; the gene size was 822 bp. Expression was also demonstrated at the level of cDNA. Also, this study revealed that pCI-S4 DNA vaccine induces TH1\TH2 response in immunized mice; this response prevents partially developing a dermal lesion of Leishmania.

Keywords: ribosomal protein S4, DNA vaccine, Leishmania tropica, BALB\c

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Authors: Merry Meryam Martgrita, Marselina Irasonia Tan

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One of several aggresive cancer is cancer that overexpress c-erbB2 receptor along with the expression of estrogen receptor. Components of extracellular matrix play an important role to increase cancer cells proliferation, migration and invasion. Both components can affect cancer development by regulating the signal transduction pathways in cancer cells. In recent research, SKOV-3 ovarian cancer cell line, that overexpress c-erbB2 receptor was cultured on type IV collagen and treated with estradiol-17β, to reveal the role of both components on RNA and protein level of c-erbB2 receptor. In this research we found a modulation phenomena of increasing and decreasing of c-erbB2 RNA level and a stabilisation phenomena of c-erbB2 protein expression due to estradiol-17β and type IV collagen. It seemed that estradiol-17β has an important role to increase c-erbB2 transcription and the stability of c-erbB2 protein expression. Type IV collagen has an opposite role. It blocked c-erbB2 transcription when it bound to integrin receptor in SKOV-3 cells.

Keywords: c-erbB2, estradiol-17β, SKOV-3, type IV collagen

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Authors: Tahra ElObeid, Omnya Ahmed, Reem Al-Sharshani, Doaa Dalloul, Jannat Alnattei

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Background: Camelus dormedarius camels are also called ‘the Arabian camels’ and are present in the desert area of North Africa and the Middle East. Recently, camel’s milk has a great attention globally because of their proteins and peptides that have been reported to be beneficial for the health and in the management of many diseases. Objectives: This study was designed to investigate the antioxidant, antimicrobial activity and to evaluate the total phenolic content of camel’s milk proteins in Qatar. Methods: Fresh two camel’s milk samples from Omani breed and called Muhajer (camel’s milk A and B) were collected on the 1st of the December. Both samples were from the same location Al- Shahaniyah, Doha, Qatar, but from different local private farms and feeding system. Camel’s milk A and B were defatted by centrifugation and their proteins were extracted by acid and thermal precipitation. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic compound (TPC) was evaluated by Folin-Ciocalteu reagent (FCR). On the other hand, the antimicrobial activity against eight different type of pathogenic bacteria was evaluated by disc diffusion method and the zone of inhibition was measured. Results: The of the total phenolic content of whole milk in both camel’s milk A and B were significantly the highest among the protein extracts. The % of the DPPH radical inhibition of casein protein in both camel’s milk A and B were significantly the highest among the protein extracts. In this study, there were marked changes in the antibacterial activity in the different camel milk protein extracts. All extracts showed bacterial overgrowth. Conclusion: The antioxidant activity of the camel milk protein extracts correlated to their unique phenolic compounds and bioactive protein peptides. The antimicrobial activity was not detected perhaps due to the technique, the quality, or the extraction method. Overall, camel's milk exhibits a high antioxidant activity, which is responsible for many health benefits besides the nutritional values.

Keywords: camels milk, antioxidant content, antimicrobial activity, proteins, Qatar

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Authors: Kuladip Jana, Bhaswati Banerjee, Parimal C. Sen

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Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems along with carcinogenesis in skin, prostate, ovary, lung and mammary glands. Our study was focused on elucidating the molecular mechanism of B(a)P induced male reproductive toxicity and its prevention with phytochemical like resveratrol. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of Wistar rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased Reactive Oxygen Species (ROS), altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38 mitogen activated protein kinase (p38MAPK), cytosolic translocation of cytochrome-c, upregulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of poly ADP ribose polymerase (PARP) and down regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like steroidogenic acute regulatory protein (StAR), cytochrome P450 IIA1 (CYPIIA1), 3β hydroxy steroid dehydrogenase (3β HSD), 17β hydroxy steroid dehydrogenase (17β HSD) showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis. Next we investigated the role of resveratrol on B(a)P induced male reproductive toxicity. Our study highlighted that resveratrol co-treatment with B(a)P maintained testicular redox potential, increased serum testosterone level and prevented steroidogenic dysfunction with enhanced expression of major testicular steroidogenic proteins (CYPIIA1, StAR, 3β HSD,17β HSD) relative to treatment with B(a)P only. Resveratrol suppressed B(a)P-induced testicular activation of p38 MAPK, ATF2, iNOS and ROS production; cytosolic translocation of Cytochome c and Caspase 3 activation thereby prevented oxidative stress of testis and inhibited apoptosis. Resveratrol co-treatment also decreased B(a)P-induced AhR protein level, its nuclear translocation and subsequent CYP1A1 promoter activation, thereby decreased protein and mRNA levels of testicular cytochrome P4501A1 (CYP1A1) and prevented BPDE-DNA adduct formation. Our findings cumulatively suggest that resveratrol prevents activation of B(a)P by modulating the transcriptional regulation of CYP1A1 and acting as an antioxidant thus prevents B(a)P-induced oxidative stress and testicular apoptosis.

Keywords: benzo(a)pyrene, resveratrol, testis, apoptosis, cytochrome P450 1A1 (CYP1A1), aryl hydrocarbon receptor (AhR), p38 MAPK/ATF2/iNOS

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Authors: Ravinder Singh, Ankita Gurao, Saroj Bandhan, Sudhir Kumar Kashyap

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The Bos taurus Beta defensin 3 is a defensin peptide secreted by neutrophils and epithelial that exhibits anti-microbial activity. It is one of the crucial components forming an innate defense against intra mammary infections in livestock. The beta defensin 3 by virtue of its anti-microbial activity inhibits major mastitis pathogens including Staphylococcus aureus and Pseudomonas aeruginosa etc, which are also responsible for biofilm formation leading to antibiotic resistance phenomenon. Therefore, the defensin may prove as a non-conventional option to treat mastitis. In this study, computational analysis has been performed including sequence comparison among species and homology modeling of Bos taurus beta defensin 3 protein. The assessments of protein structure were done using the protein structure and model assessment tools integrated in Swiss Model server, which employs various local and global quality evaluation parameters. Further, molecular docking was also carried out between the defensin peptide and the components of biofilm to gain insight into various interactions and structural differences crucial for functionality of this protein.

Keywords: beta defensin 3, bos taurus, docking, homology modeling

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Authors: Mariia A. Parfenenko, Ilya S. Dantsev, Sergei V. Bochenkov, Natalia V. Vinogradova, Olga S. Groznova, Victoria Yu. Voinova

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Introduction: Autism is the most common neurodevelopmental disorder. Autism is characterized by difficulties in social interaction and adherence to stereotypic behavioral patterns and frequently co-occurs with epilepsy, intellectual disabilities, connective tissue disorders, and other conditions. CHD8 codes for chromodomain-helicase-DNA-binding protein 8 - a chromatin remodeler that regulates cellular proliferation and neurodevelopment in embryogenesis. CHD8 is one of the genes most frequently involved in autism. Patients and methods: 2 unrelated male patients, P3 and P12, aged 3 and 12 years old, underwent whole genome sequencing, which determined that they both had different likely pathogenic variants, both previously undescribed in literature. Sanger sequencing later determined that P12 inherited the variant from his affected mother. Results: P3 and P12 presented with autism, a developmental delay, ataxia, sleep disorders, overgrowth, and macrocephaly, as well as other clinical features typically present in patients with causative variants in CHD8. The mother of P12 also has autistic traits, as well as ataxia, hypotonia, sleep disorders, and other symptoms. However, P3 and P12 also have different cardiac abnormalities. P3 had signs of a repolarization disorder: a flattened T wave in the III and aVF derivations and a negative T wave in the V1-V2 derivations. He also had structural valve anomalies with associated regurgitation, local contractility impairment of the left ventricular, and diastolic dysfunction of the right ventricle. Meanwhile, P12 had Wolff-Parkinson-White syndrome and underwent radiofrequency ablation at the age of 2 years. At the time of observation, P12 had mild sinus arrhythmia and an incomplete right bundle branch block, as well as arterial hypertension. Discussion: Cardiac abnormalities were not previously reported in patients with causative variants in CHD8. The underlying mechanism for the formation of those abnormalities is currently unknown. However, the two hypotheses are either a disordered interaction with CHD7 – another chromodomain remodeler known to be directly involved in the cardiophenotype of CHARGE syndrome – a rare condition characterized by coloboma, heart defects and growth abnormalities, or the disrupted functioning of CHD8 as an A-Kinase Anchoring Protein, which are known to modulate cardiac function. Conclusion: We observed 2 unrelated autistic males with likely pathogenic variants in CHD8 that presented with typical symptoms of CHD8-related neurodevelopmental disorder, as well as cardiac abnormalities. Cardiac abnormalities have, until now, been considered uncharacteristic for patients with causative variants in CHD8. Further accumulation of data, including experimental evidence of the involvement of CHD8 in heart formation, will elucidate the mechanism underlying the cardiophenotype of those patients. Acknowledgements: Molecular genetic testing of the patients was made possible by the Charity Fund for medical and social genetic aid projects «Life Genome.»

Keywords: autism spectrum disorders, chromodomain-helicase-DNA-binding protein 8, neurodevelopmental disorder, cardio phenotype

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Authors: R. Sabre

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This work focuses on the symmetric alpha stable process with continuous time frequently used in modeling the signal with indefinitely growing variance, often observed with an unknown additive error. The objective of this paper is to estimate this error from discrete observations of the signal. For that, we propose a method based on the smoothing of the observations via Jackson polynomial kernel and taking into account the width of the interval where the spectral density is non-zero. This technique allows avoiding the “Aliasing phenomenon” encountered when the estimation is made from the discrete observations of a process with continuous time. We have studied the convergence rate of the estimator and have shown that the convergence rate improves in the case where the spectral density is zero at the origin. Thus, we set up an estimator of the additive error that can be subtracted for approaching the original signal without error.

Keywords: spectral density, stable processes, aliasing, non parametric

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Authors: Nadereh Sohrabi Shegefti, Siamak Samani

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Level of adjustment is the first index to check mental health. The aim of this study was developing a valid and reliable Multidimensional Adjustment Scale (MAS). The sample consisted of 150 college students. Multidimensional adjustment scale and Depression, Anxiety, and stress scale (DASS) were used in this study. Principle factor analysis, Pearson correlation coefficient, and Cornbach's Alpha were used to check the validity and reliability of the MAS. Principle component factor analysis showed a 5 factor solution for the MAS. Alpha coefficients for the MAS sub scales were ranged between .69 to .83. Test-retest reliability for MAS was .88 and the mean of sub scales- total score correlation was .88. All these indexes revealed an acceptable reliability and validity for the MAS. The MAS is a short assessment instrument with good acceptable psychometric properties to use in clinical filed.

Keywords: psychological adjustment, psychometric properties, validity, Pearson correlation

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Authors: Ahreum Choi, Otgontuya Tsogbadrakh, Kwang-Hwan Jung

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Microbial rhodopsins are well-known seven-transmembrane proteins that have been extensively studied. These retinal-binding proteins have divided into two types. The type I is microbial rhodopsin, and type II (visual pigment) is expressed mostly in mammalian eyes. For type I rhodopsin, there are two main functions that are ion pumping activity and sensory transduction. Anabaena sensory rhodopsin (ASR) is one of the microbial rhodopsin with main function as photo-sensory transduction. Although ASR is expressed fairly well in Escherichia coli, the expression level is relatively less compare to Proteorhodopsin. In this study, full length of ASR was used to test for the expression influence by codon usage in E. coli. Eight amino acids of codon at N-terminal part of ASR were changed randomly with designed primers, which allow 8,192 nucleotide different cases. The codon changes were screened for the preferable codons of each residue, which have given higher expression yield. Among those 57 selected mutations, there are 24 color-enhanced E. coli colonies that contain ASR proteins, and it showed better expression level than the wild type ASR codon usage. This strongly suggests that high codon usage of only partial N-terminal of protein can increase the expression level of whole protein.

Keywords: 7-transmembrane, all-trans retinal, rhodopsin, codon-usage, protein expression

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Authors: Enjie Xu, Zhen Huang, Kunpeng Zhu, Jianping Hu, Xiaolong Ma, Yongjie Wang, Jiazhuang Zhu, Chunlin Zhang

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Abstract: Hypoxia and dedifferentiation of osteosarcoma (OS) cells leads to poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated signaling pathways. We performed a sphere formation assay and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the expression relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and the effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS metastasis. Dedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A promoted the expression of PDGFRB, subsequently activated ras homolog family member A (RhoA), and increased the phosphorylation of myosin light chain (MLC). PDGFRB also enhanced the phosphorylation of focal adhesion kinase (FAK). The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB also promoted cell dedifferentiation and had a significant impact on the metastasis of OS cells both in vitro and in vivo. Our results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton by activating RhoA and subsequently phosphorylating MLC, thereby promoting OS dedifferentiation and pulmonary metastasis.

Keywords: osteosarcoma, dedifferentiation, metastasis, cytoskeleton rearrangement, PDGFRB, hypoxia

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Authors: Vijay Kumar Kutala, Addepalli Pavani, M. Amresh Rao, Naushad Sm

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In this study, we evaluated the impact of CYP2C9*2 and CYP2C9*3 variants on binding and hydroxylation of warfarin. In silico data revealed that warfarin forms two hydrogen bonds with protein backbone i.e. I205 and S209, one hydrogen bond with protein side chain i.e. T301 and stacking interaction with F100 in CYP2C9*1. In CYP2C9*2 and CYP2C9*3 variants, two hydrogen bonds with protein backbone are disrupted. In double variant, all the hydrogen bonds are disrupted. The distances between C7 of S-warfarin and Fe-O in CYP2C9*1, CYP2C9*2, CYP2C9*3 and CYP2C9*2/*3 were 5.81A°, 7.02A°, 7.43° and 10.07°, respectively. The glide scores (Kcal/mol) were -7.698, -7.380, -6.821 and -6.986, respectively. Increase in warfarin/7-hydroxy warfarin ratio was observed with increase in variant alleles. To conclude, CYP2C9*2 and CYP2C9*3 variants result in disruption of hydrogen bonding interactions with warfarin and longer distance between C7 and Fe-O thus impairing warfarin 7-hydroxylation due to lower binding affinity of warfarin.

Keywords: warfarin, CYP2C9 polymorphism, personalized medicine, in Silico

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Authors: Eleni Poulou-Sidiropoulou, Charalampos N. Bompas, Martina Samiotaki, Alexios Vlamis-Gardikas

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The glutaredoxin (Grx) and thioredoxin (Trx) systems keep the intracellular environment reduced in almost all organisms. In Escherichia coli (E. coli), the Grx system relies on NADPH+ to reduce GSH reductase (GR), the latter reducing oxidized diglutathione to glutathione (GSH) which in turn reduces cytosolic Grxs, the electron donors for different intracellular substrates. In the Trx system, GR and GSH are replaced by Trx reductase (TrxR). Three of the Grxs of E. coli (Grx1, 2, 3) are reduced by GSH, while Grx4 is likely reduced by TrxR. Trx1 and Grx1 from E. coli may reduce ribonucleotide reductase Ia to ensure a constant supply of deoxyribonucleotides for the synthesis of DNA. The role of the other three Grxs is relatively unknown, especially for Grx2 that may amount up to 1 % of total cellular protein in the stationary phase of growth. The protein is known as a potent antioxidant, but no specific functions have been attributed to it. Herein, affinity chromatography of cellular extracts on immobilized Grx2, followed by MS analysis of the resulting eluates, was employed to identify protein ligands that could provide insights into the biological role of Grx2. Ionic, strong non-covalent, and covalent (disulfide) interactions with relevant proteins were detected. As a means of verification, the identified ligands were subjected to in silico docking with monothiol Grx2. In other experiments, protein extracts from E. coli cells lacking the gene for Grx2 (grxB) were compared to those of wild type. Taken together, the two approaches suggest that Grx2 is involved in protein synthesis, nucleotide metabolism, DNA damage repair, stress responses, and various metabolic processes. Grx2 appears as a versatile protein that may participate in a wide range of biological pathways beyond its known general antioxidant function.

Keywords: Escherichia coli, glutaredoxin 2, interactome, thiol-disulfide oxidoreductase

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Authors: Shiva Emami, Jenny Persson, Bengt Johansson Lindbom

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Group A streptococcus (GAS) is a prevalent human pathogen, causing a wide range of infections and diseases. One of the most well-known virulence factors in GAS is M protein, a surface protein that facilitates bacterial invasion. In this study, we used a recombinant GAS strain (GAS-2W) expressing M protein containing a hyper immunogenic peptide (2W). Mice were immunized three times with heat-killed-GAS subcutaneously at three weeks intervals. Three weeks post last immunization, mice were challenged intraperitoneally with a lethal dose of live GAS. In order to investigate the impact of IFN-ƴ and antibodies in protection against GAS infection, we used a mouse model knock-out for IFN-ƴ (IFN-ƴ KO). We observed immunization with GAS-2W strain can increase protection against GAS infection in mice compared with the original GAS strain. Higher levels of antibodies against M1 protein were measured in GAS-2W-immunized mice. There was also a significant increase in IgG2c response in mice immunized with GAS2W. By using IFN-ƴ KO mice, we showed that not a high level of total IgG, but IgG2c was correlated with protection through the i.p challenge. It also emphasizes the importance of IFN-ƴ cytokine to combat GAS by isotype switching to IgG2c (which is opsonic for phagocytosis). Our data indicate the crucial role of IFN-ƴ in the protective immune response that, together with IgG2c, can induce protection against GAS.

Keywords: Group A streptococcus, IgG2c, IFN-γ, protection

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Authors: Ravinder Singh, C Rajesh, Saroj Badhan, Shailendra Mishra, Ranjit Singh Kataria

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Heat shock protein 60 and 70 are crucial chaperones that guide appropriate folding of denatured proteins under heat stress conditions. HSP60 and HSP70 provide assistance in correct folding of a multitude of denatured proteins. The heat shock factors are the family of some transcription factors which controls the regulation of gene expression of proteins involved in folding of damaged or improper folded proteins during stress conditions. Under normal condition heat shock proteins bind with HSF-1 and act as its repressor as well as aids in maintaining the HSF-1’s nonactive and monomeric confirmation. The experimental protein structure for all these proteins in Bubalus bubalis is not known till date. Therefore computational approach was explored to identify three-dimensional structure analysis of all these proteins. In this study, an extensive in silico analysis has been performed including sequence comparison among species to comparative modeling of Bubalus bubalis HSP60, HSP70 and HSF-1 protein. The stereochemical properties of proteins were assessed by utilizing several scrutiny bioinformatics tools to ensure model accuracy. Further docking approach was used to study interactions between Heat shock proteins and HSF-1.

Keywords: Bubalus bubalis, comparative modelling, docking, heat shock protein

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Authors: Farnaz Firuzian, Farhad Choobdar, Ali Mazouri

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As Very Low Birth Weight (VLBW) neonates cannot tolerate enteral feeding, parenteral nutrition (PN) must be administered shortly after birth. To find an optimal combination of nutrition, in this study, we compare administering high glucose versus high protein intake as a component of total parenteral nutrition (TPN) to test their effect on birth weight (BW) regain in VLBW. This study employs a multi-arm randomized trial: 145 newborns with BW < 1500 g were randomized to control (C) or experimental groups: high glucose (G) or high protein (P). All samples in each group received the same TPN regimens except glucose and protein intake: Glocuse was provided by dextrose water (DW) serum: 7-15 g/kg/d (10% DW) in groups C and P versus 8.75-18.75 g/kg/d (12.5% DW) in group G. Protein provided by amino acids 3 g/kg/d for groups C and G versus 4 g/kg/d for group P. Outcomes (weight, height, and head circumference) was monitored on a daily basis until the BW was regained. Data has been gathered recently and is being processed. We hypothesize that neonates with higher amino acid intake will result in sooner BW regain than other groups. The result will be presented at the conference. The findings of this study not only can help optimize nutrition, cost reduction, and shorter NICU admission of VLBW neonates at the hospital level but eventually contribute to reduced healthcare-associated infections (HAIs) and an improved health economy.

Keywords: very low birth weight neonates, weight gain, parenteral nutrition, glucose, amino acids

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Authors: Lukas Stiburek, Jana Cesnekova, Josef Houstek, Jiri Zeman

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Cellular function depends on mitochondrial function and integrity that is therefore maintained by several classes of proteins possessing chaperone and/or proteolytic activities. In this work, we focused on characterization of LACE1 (lactation elevated 1) function in mitochondrial protein homeostasis maintenance. LACE1 is the human homologue of yeast mitochondrial Afg1 ATPase, a member of SEC18-NSF, PAS1, CDC48-VCP, TBP family. Yeast Afg1 was shown to be involved in mitochondrial complex IV biogenesis, and based on its similarity with CDC48 (p97/VCP) it was suggested to facilitate extraction of polytopic membrane proteins. Here we show that LACE1, which is a mitochondrial integral membrane protein, exists as part of three complexes of approx. 140, 400 and 500 kDa and is essential for maintenance of fused mitochondrial reticulum and lamellar cristae morphology. Using affinity purification of LACE1-FLAG expressed in LACE1 knockdown background we show that the protein physically interacts with mitochondrial inner membrane protease YME1L. We further show that human LACE1 exhibits significant pro-apoptotic activity and that the protein is required for normal function of the mitochondrial respiratory chain. Thus, our work establishes LACE1 as a novel factor with the crucial role in mitochondrial homeostasis maintenance.

Keywords: LACE1, mitochondria, apoptosis, protease

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Authors: Hamza Javar Magnier, Robin Curtis

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There have been rapid advances in the upstream processing of protein therapeutics, which has shifted the bottleneck to downstream purification and formulation. Finding liquid formulations with shelf lives of up to two years is increasingly difficult for some of the newer therapeutics, which have been engineered for activity, but their formulations are often viscous, can phase separate, and have a high propensity for irreversible aggregation1. We explore means to develop improved predictive ability from a better understanding of how protein-protein interactions on formulation conditions (pH, ionic strength, buffer type, presence of excipients) and how these impact upon the initial steps in protein self-association and aggregation. In this work, we study the initial steps in the aggregation pathways using a minimal protein model based on square-well potentials and discontinuous molecular dynamics. The effect of model parameters, including range of interaction, stiffness, chain length, and chain sequence, implies that protein models fold according to various pathways. By reducing the range of interactions, the folding- and collapse- transition come together, and follow a single-step folding pathway from the denatured to the native state2. After parameterizing the model interaction-parameters, we developed an understanding of low-temperature conformational properties and fluctuations, and the correlation to the folding transition of proteins in isolation. The model fluctuations increase with temperature. We observe a low-temperature point, below which large fluctuations are frozen out. This implies that fluctuations at low-temperature can be correlated to the folding transition at the melting temperature. Because proteins “breath” at low temperatures, defining a native-state as a single structure with conserved contacts and a fixed three-dimensional structure is misleading. Rather, we introduce a new definition of a native-state ensemble based on our understanding of the core conservation, which takes into account the native fluctuations at low temperatures. This approach permits the study of a large range of length and time scales needed to link the molecular interactions to the macroscopically observed behaviour. In addition, these models studied are parameterized by fitting to experimentally observed protein-protein interactions characterized in terms of osmotic second virial coefficients.

Keywords: protein folding, native-ensemble, conformational fluctuation, aggregation

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Authors: Oqba Basal, Andras Szabo

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Using mineral fertilization is increasing worldwide, as it is claimed to be majorly responsible for achieving high yields; however, the negative impacts of mineral fertilization on soil and environment are becoming more obvious, with alternative methods being more necessary and applicable, especially with the current climatic changes which have imposed serious abiotic stresses, such as drought. An experiment was made during 2017 growing season in Debrecen, Hungary to investigate the effects of inoculation and N fertilization on the seed yield and protein concentration of the soybean (Glycine max (L.) Merr.) cultivar (Panonia Kincse) under three different irrigation regimes: severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Three N fertilizer rates were applied: no N fertilizer (0 N), 35 kg ha⁻¹ of N fertilizer (35 N) and 105 kg ha⁻¹ of N fertilizer (105 N). Half of the seeds in each treatment was inoculated with Bradyrhizobium japonicum inoculant, and the other half was not inoculated. The results showed significant differences in the seed yield associated with inoculation, irrigation and the interaction between them, whereas there were no significant differences in the seed yield associated with fertilization alone or in interaction with inoculation or irrigation or both. When seeds were inoculated, yield was increased when (35 N) was applied compared to (0 N) but not significantly; however, the high rate of N fertilizer (105 N) reduced the yield to a level even less than (0 N). When seeds were not inoculated, the highest rate of N increased the yield the most compared to the other two N fertilizer rates whenever the drought was present (moderate or severe). Under severe drought stress, inoculation was positively and significantly correlated with yield; however, adding N fertilizer increased the yield of uninoculated plants compared to the inoculated ones, regardless of the rate of N fertilizer. Protein concentration in the seeds was significantly affected by irrigation and by fertilization, but not by inoculation. Protein concentration increased as the N fertilization rate increased, regardless of the inoculation or irrigation treatments; moreover, increasing the N rate reduced the correlation coefficient of protein concentration with the irrigation. It was concluded that adding N fertilizer is not always recommended, especially when seeds are inoculated before being sown; however, it is very important under severe drought stress to sustain yield. Enhanced protein concentrations could be achieved by applying N fertilization, whether the seeds were pre-inoculated or not.

Keywords: drought stress, N fertilization, protein concentration, soybean

Procedia PDF Downloads 132

Authors: Richa Dhingra, Monika, Manav Malhotra, Tilak Raj Bhardwaj, Neelima Dhingra

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5-Alpha-reductases (5AR), a membrane bound, NADPH dependent enzyme and convert male hormone testosterone (T) into more potent androgen dihydrotestosterone (DHT). DHT is the required for the development and function of male sex organs, but its overproduction has been found to be associated with physiological conditions like Benign Prostatic Hyperplasia (BPH). Thus the inhibition of 5ARs could be a key target for the treatment of BPH. In present study, 2D and 3D Quantitative Structure Activity Relationship (QSAR) pharmacophore models have been generated for 5AR based on known inhibitory concentration (IC₅₀) values with extensive validations. The four featured 2D pharmacophore based PLS model correlated the topological interactions (–OH group connected with one single bond) (SsOHE-index); semi-empirical (Quadrupole2) and physicochemical descriptors (Mol. wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity, and has the highest correlation coefficient (r² = 0.98, q² =0.84; F = 57.87, pred r² = 0.88). Internal and external validation was carried out using test and proposed set of compounds. The contribution plot of electrostatic field effects and steric interactions generated by 3D-QSAR showed interesting results in terms of internal and external predictability. The well validated 2D Partial Least Squares (PLS) and 3D k-nearest neighbour (kNN) models were used to search novel 5AR inhibitors with different chemical scaffold. To gain more insights into the molecular mechanism of action of these steroidal derivatives, molecular docking and in silico absorption, distribution, metabolism, and excretion (ADME) studies were also performed. Studies have revealed the hydrophobic and hydrogen bonding of the ligand with residues Alanine (ALA) 63A, Threonine (THR) 60A, and Arginine (ARG) 456A of 4AT0 protein at the hinge region. The results of QSAR, molecular docking, in silico ADME studies provide guideline and mechanistic scope for the identification of more potent 5-Alpha-reductase inhibitors (5ARI).

Keywords: 5α-reductase inhibitor, benign prostatic hyperplasia, ligands, molecular docking, QSAR

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Authors: Medha Basu, Kumardeb Banerjee, Dipak Ghosh

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After the massive blow of the COVID-19 pandemic, several health hazards have been reported all over the world. Serious cases of Major Depressive Disorder (MDD) are seen to be common in about 15% of the global population, making depression one of the leading mental health diseases, as reported by the World Health Organization. Various psychological and pharmacological treatment techniques are regularly being reported. Music, a globally accepted mode of entertainment, is often used as a therapeutic measure to treat various health conditions. We have tried to understand how Indian Classical Music can affect the overall well-being of the human brain. A case study has been reported here, where a Flute-rendition has been chosen from a detailed audience response survey, and the effects of that clip on human brain conditions have been studied from both psychological and neural perspectives. Taking help from internationally-accepted depression-rating scales, two questionnaires have been designed to understand both the prolonged and immediate effect of music on various emotional states of human lives. Thereafter, from EEG experiments on 5 participants using the same clip, the parameter ‘ALAY’, alpha frontal asymmetry (alpha power difference of right and left frontal hemispheres), has been calculated. Works of Richard Davidson show that an increase in the ‘ALAY’ value indicates a decrease in depressive symptoms. Using the non-linear technique of MFDFA on EEG analysis, we have also calculated frontal asymmetry using the complexity values of alpha-waves in both hemispheres. The results show a positive correlation between both the psychological survey and the EEG findings, revealing the prominent role of music on the human brain, leading to a decrease in mental unrest and an increase in overall well-being. In this study, we plan to propose the scientific foundation of music therapy, especially from a neurocognition perspective, with appropriate neural bio-markers to understand the positive and remedial effects of music on the human brain.

Keywords: music therapy, EEG, psychological survey, frontal alpha asymmetry, wellbeing

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Authors: Sarita Puri, Tapan K. Chaudhuri

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Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.

Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability

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Authors: Saba Belgheisi

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Food packaging, is an important discipline in the area of food technology, concerns preservation and protection of foods. The objective of this research was to determine of the effect of whey protein based edible coating on the moisture loss and sensory attributes of fresh mutton after 0, 1, 3 and 5 days at 5° C. The moisture content, moisture loss and sensory attributes (juiciness, color and odor) of the coated and uncoated samples were analyzed. The results showed that, moisture content, moisture loss, juiciness and color of the coated and uncoated samples have significant differences (p < 0.05) at the intervals of 0 to 1 and 1 to 3 days of storage. But no significant difference was observed at interval time 3 to 5 days of storage (p > 0.05). Also, there was no significant differences in the odor values of the coated and uncoated samples (p > 0.05). Therefore, the coated samples had consistently more moisture, juiciness and colored values than uncoated samples after 3 days at 5° C. So, whey protein edible coating could enhance product presentation and eliminate the need for placing absorbent pads at the bottom of the trays.

Keywords: coating, whey protein, mutton, moisture, sensory

Procedia PDF Downloads 443

Authors: Sodamade A., Bolaji K. A.

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Problems associated with protein malnutrition are still prevalent in third-world countries, leading to the constant search for plants that could serve as nutrients and medicinal purposes. Huckleberry is one of the plants that has been proven useful locally in the treatment of numerous ailments and diseases. A fresh sample of the plant (Vaccinium pavifolium) was collected from a vegetable garden situated near the Erelu dam of the Emmanuel Alayande College of Education Campus, Oyo. The sample was authenticated at the Forestry Research Institute of Nigeria (FRIN) Ibadan. The leaves of the plant were plucked and processed for leaf protein concentrates before proximate composition, mineral analysis phytochemical and antimicrobial properties were determined using a standard method of analysis. The results of proximate constituents showed; moisture content; 9.89±0.051g/100g, Ash; 3.23±0.12g/100g, crude fat; 3.96±0.11g/100g and 61.27±0.56g/100g of Nitrogen free extractive. The mineral analysis of the sample showed; Mg; 0.081±0.00mg/100g, Ca; 42.30±0.05mg/100g, Na; 27.57±0.09mg/100g, K; 6.81±0.01mg/100g, P; 8.90±0.03mg/100g, Fe; 0.51±0.00mg/100g, Zn; 0.021±0.00mg/100g, Cd; 0.04±0.04mg/100g, Pb; 0.002±0.00mg/100g, Cr; 0.041±0.00mg/100g Cadmium and Mercury were not detected in the sample. The result of phytochemical analysis of leaf protein concentrates of the Huckleberry showed the presence of Alkaloid, Saponin, Flavonoid, Tanin, Coumarin, Steroids, Terpenoids, Cardiac glycosides, Glycosides, Quinones, Anthocyanin, phytosterols, and phenols. Ethanolic extracts of the Vaccinium parvifolium L. leaf protein concentrates showed that it contains bioactive compounds that are capable of combating the following microorganisms; Staphylococcus aureus, Streptococcus pyogenes, Streptococcus faecalis, Pseudomonas aeruginosa, Klebisialae pneumonia and Proteus mirabilis. The results of the analysis of Vaccinium parvifolium L. leaf protein concentrates showed that the sample contains valuable nutrient and mineral constituents, and phytochemical compounds that could make the sample useful for medicinal activities.

Keywords: leaf protein concentrates, vaccinium parvifolium, nutritional characteristics, mineral composition, antimicrobial activity

Procedia PDF Downloads 52

Authors: Hemalatha Ganapathyswamy, Thirukkumar Subramani

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Celiac disease is an autoimmune response to dietary wheat gluten. Gluten is the main structure forming protein in bread and hence developing gluten-free bread is a technological challenge. The study aims at using nonwheat flours like barnyard millet and lentil flour to replace wheat in bread formulations. Other characteristics of these grains, such as high protein, soluble fiber, mineral content and bioactive components make them attractive alternatives to traditional gluten-free ingredients in the production of high protein, gluten-free bread. The composite flour formulations for the development of gluten-free bread were optimized using lentil flour (50 to 70 g), barnyard millet flour (0 to 30 g) and corn flour (0 to 30 g) by means of response surface methodology with various independent variables for physical, sensorial and nutritional characteristics. The optimized composite flour which had a desirability value of 0.517, included lentil flour –62.94 g, barnyard millet flour– 24.34 g and corn flour– 12.72 g with overall acceptability score 8.00/9.00. The optimized gluten-free bread formulation had high protein (14.99g/100g) and fiber (1.95g/100g) content. The glycemic index of the gluten-free bread was 54.58 rendering it as low glycemic which enhances the functional benefit of the gluten-free bread. Since the standardised gluten-free bread from barnyard millet and lentil flour are high protein, and gluten-free with low glycemic index, the product would serve as an ideal therapeutic food in the management of both celiac disease and diabetes mellitus with better nutritional value.

Keywords: gluten free bread, lentil, low glycemic index, response surface methodology

Procedia PDF Downloads 168