Search results for: protein Z

Authors: Strahinja Kovačević, Lidija Jevrić, Miloš Kuzmanović, Sanja Podunavac-Kuzmanović

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In this paper, various derivatives of benzimidazole have been evaluated against Gram-negative bacteria Escherichia coli. For all investigated compounds the minimum inhibitory concentration (MIC) was determined. Quantitative structure-activity relationships (QSAR) attempts to find consistent relationships between the variations in the values of molecular properties and the biological activity for a series of compounds so that these rules can be used to evaluate new chemical entities. The correlation between MIC and some absorption, distribution, metabolism and excretion (ADME) parameters was investigated, and the mathematical models for predicting the antibacterial activity of this class of compounds were developed. The quality of the multiple linear regression (MLR) models was validated by the leave-one-out (LOO) technique, as well as by the calculation of the statistical parameters for the developed models and the results are discussed on the basis of the statistical data. The results of this study indicate that ADME parameters have a significant effect on the antibacterial activity of this class of compounds. Principal component analysis (PCA) and agglomerative hierarchical clustering algorithms (HCA) confirmed that the investigated molecules can be classified into groups on the basis of the ADME parameters: Madin-Darby Canine Kidney cell permeability (MDCK), Plasma protein binding (PPB%), human intestinal absorption (HIA%) and human colon carcinoma cell permeability (Caco-2).

Keywords: benzimidazoles, QSAR, ADME, in silico

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Authors: Abderrahim Cheriguene, Fatiha Arioui

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The experimental study focuses on the extraction of pectin, whey protein and gelatin, and the study of their functional properties. Microbiological, physicochemical and sensory approach integrated has been implanted to study the effect of the incorporation of these natural food additives in the matrix of a fermented milk type set yogurt, to study the stability of the product during the periods of fermentation and post-acidification over a period of 21 days at 4°C. Pectin was extracted in hot HCl solution. Thermo-precipitation was carried out to obtain the whey proteins while the gelatin was extracted by hydrolysis of the collagen from bovine ossein. The fermented milk was prepared by varying the concentration of the incorporated additives. The measures and controls carried performed periodically on fermented milk experimental tests were carried out: pH, acidity, viscosity, the enumeration of Streptococcus thermophilus, cohesiveness, adhesiveness, taste, aftertaste, whey exudation, and odor. It appears that the acidity, viscosity, and number of Streptococcus thermophilus increased with increasing concentration of additive added in the experimental tests. Indeed, it seems clear that the quality of fermented milk and storability is more improved than the incorporation rate is high. The products showed a better test and a firmer texture limiting the whey exudation.

Keywords: fermented milk, pectin, gelatin, whey proteins, functional properties, quality, conservation, valorization

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Authors: Dominika Pihikova, Stefan Belicky, Tomas Bertok, Roman Sokol, Petra Kubanikova, Jan Tkac

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Since aberrant glycosylation is frequently accompanied by both physiological and pathological processes in a human body (cancer, AIDS, inflammatory diseases, etc.), the analysis of tumor-associated glycan patterns have a great potential for the development of novel diagnostic approaches. Moreover, altered glycoforms may assist as a suitable tool for the specificity and sensitivity enhancement in early-stage prostate cancer diagnosis. In this paper we discuss the construction and optimization of ultrasensitive sandwich biosensor platform employing lectin as glycan-binding protein. We focus on the immunoassay development, reduction of non-specific interactions and final glycoprofiling of human serum samples including both prostate cancer (PCa) patients and healthy controls. The fabricated biosensor was measured by label-free electrochemical impedance spectroscopy (EIS) with further lectin microarray verification. Furthermore, we analyzed different biosensor interfaces with atomic force microscopy (AFM) in nanomechanical mapping mode showing a significant differences in the altitude. These preliminary results revealing an elevated content of α-2,3 linked sialic acid in PCa patients comparing with healthy controls. All these experiments are important step towards development of point-of-care devices and discovery of novel glyco-biomarkers applicable in cancer diagnosis.

Keywords: biosensor, glycan, lectin, prostate cancer

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Authors: Zhang Zhenzhen

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Valproate (VPA) is commonly used in the treatment of bipolar disorder and epilepsy. The mechanism is complicated, including its ability to inhibit histone deacetylases (HDACs). Here, we show that VPA attenuated VEGF gene expression and the morphological changes in choroidal neovascularization (CNV) induced by photocoagulation in retina. C57BL/6 mice were injected subcutaneously at 300mg/kg twice daily with VPA before insult. Vascular endothelial growth factor (VEGF)-A and VEGF-B were examined in the eyes of VPA-treated mice and in human retinal pigment epithelial cell lines (ARPE-19) exposed to VPA. In addition, CNV was induced by photocoagulation in mice injected with VPA, and the volume of CNV was compared by fluorescence-labeled choroidal flat mount. Morphological changes were analyzed on stained histological sections. Western blot analysis was used to determine protein levels of VEGF-A and VEGF-B, and acetylation of histone H3 in each group. VPA injected intraperitoneally attenuated the VEGF-A and VEGF-B expression in the retina, accompanied by the hyperacetylation of retina tissue, indicating that VPA acts directly on retina tissues through acetylation to reduce the expression of VEGF. VPA also attenuated the VEGF-A mRNA expression in the retinal pigment epithelium showed by immunohistochemistry. Moreover, the administration of VPA significantly attenuated photocoagulation-induced CNV in mice. These results demonstrate that VPA attenuated VEGF production in retina associated with choroidal neovascularization possibly via the HDAC inhibition.

Keywords: retina, acetylation, chorodial neovascularization, vascular endothelial growth factor

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Authors: Aminat. O Adelekan, Olawale T. Gbadebo

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Biscuit is one of the most consumed cereal foods in Nigeria and research has shown that locally available tropical crops like cassava, sweet potato can be made into flour and used in the production of biscuits and other pastries. This study investigates some quality parameters in biscuits made from blends of wheat, cassava and carrot flour. The values of result of samples increased with increasing percentage substitution of cassava and carrot flour in some quality parameter like fiber, ash, gluten content, and carbohydrate. The protein content reduced significantly (P < 0.05) with increasing percentage substitution of cassava and carrot flour which ranged from 14.80% to 11.80% compared with the control sample which had 15.60%. There was a recorded significant increase (P < 0.05) in some mineral composition such as calcium, magnesium, sodium, iron, phosphorus, and vitamin A and C composition as the percentage substitution of cassava and carrot flour increased. During storage stability test, samples stored in the fridge and freezer were found to be the best storage location to preserve the sensory attributes and inhibit microbial growth when compared with storage under the sun and on the shelf. Biscuit made with blends of wheat, cassava and carrot flour can therefore serve as an alternative to biscuits made from 100% wheat flour, as they are richer in vitamin A, vitamin C, carbohydrate, dietary fiber and some essential minerals.

Keywords: biscuit, carrot, flour blends, storage

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Authors: Fikadu Kifle Hailegeorgis

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Common bean is one of the most important sources of protein in Ethiopia and other developing countries. However, the Mexican bean weevil, Zabrotes subfasciatus (Boheman), is a major factor in the storage of common beans that causes losses. Studies were conducted to evaluate the efficacy of botanical powders of Jatropha curcas (L.), Neem/Azadrachta indica, and Parthenium hysterophorus (L) on local common bean varieties against Z subfasciatus at Melkassa Agriculture Research Center. Twenty local common bean varieties were evaluated twice against Z. Subfasciatus in a completely randomized design in three replications at the rate of 0.2g/250g of seed for each experiment. Malathion and untreated were used as standard checks. The result indicated that RAZ White and Round Yellow showed high resistance variety in experiments while Batu and Black showed high susceptible variety in experiments. Jatropha seed powder was the most effective against Z. subfasciatus. Parthenium seed powders and neem leaf powders also indicate promising results. Common beans treated with botanicals significantly (p<0.05) had a higher germination percentage than that of the untreated seed. In general, the results obtained indicated that using bean varieties (RAZ white and Round yellow) and botanicals (Jatropha) seed powder gave the best control of Z. subfasciatus.

Keywords: botanicals, malathion, resistant varieties, Z. subfasciatus

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Authors: Rozhgar A. Khailany, Mehri Igci, Emine Bayraktar, Sakip Erturhan, Metin Karakok, Ahmet Arslan

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Kidney cancer is the most lethal urological cancer accounting for 3% of adult malignancies. VHL, a tumor-suppressor gene, is best known to be associated with renal cell carcinoma (RCC). The VHL functions as negative regulator of hypoxia inducible factors. Recent sequencing efforts have identified several novel frequent mutations of histone modifying and chromatin remodeling genes in ccRCC (clear cell RCC) including PBRM1 and SETD2. The PBRM1 gene encodes the BAF180 protein, which involved in transcriptional activation and repression of selected genes. SETD2 encodes a histone methyltransferase, which may play a role in suppressing tumor development. In this study, RNAs of 30 paired tumor and normal samples that were grouped according to the types of kidney cancer and clinical characteristics of patients, including gender and average age were examined by RT-PCR, SSCP and sequencing techniques. VHL, PBRM1 and SETD2 expressions were relatively down-regulated. However, statistically no significance was found (Wilcoxon signed rank test, p > 0.05). Interestingly, no mutation was observed on the contrary of previous studies. Understanding the molecular mechanisms involved in the pathogenesis of RCC has aided the development of molecular-targeted drugs for kidney cancer. Further analysis is required to identify the responsible genes rather than VHL, PBRM1 and SETD2 in kidney cancer.

Keywords: kidney cancer, molecular biomarker, expression analysis, mutation screening

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Authors: Xiang Wang, Yan Pang, Zhaomiao Liu

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Droplet-based microfluidics have been studied extensively with the development of the Micro-Electro-Mechanical System (MEMS) which bears the advantages of high throughput, high efficiency, low cost and low polydispersity. Droplets, worked as versatile carriers, could provide isolated chambers as the internal dispersed phase is protected from the outside continuous phase. Droplets are used to add reagents to start or end bio-chemical reactions, to generate concentration gradients, to realize hydrate crystallization or protein analyses, while droplets coalescence acts as an important control technology. In this paper, deionized water is used as the dispersed phase, and several kinds of oil are used as the continuous phase to investigate the influence of the viscosity ratio of the two phases on the coalescence process. The microchannels are fabricated by coating a polydimethylsiloxane (PDMS) layer onto another PDMS flat plate after corona treatment. All newly made microchannels are rinsed with the continuous oil phase for hours before experiments to ensure the swelling fully developed. High-speed microscope system is used to document the serial videos with a maximum speed of 2000 frames per second. The critical capillary numbers (Ca*) of droplet pairs in various junctions are studied and compared. Ca* varies with different junctions or different liquids within the range of 0.002 to 0.01. However, droplets without extra control would have the problem of synchronism which reduces the coalescence efficiency.

Keywords: coalescence, concentration, critical capillary number, droplet pair, split

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Authors: Kriti Upadhyay, Ashraf Ali, Puja Sohal, Randeep Guleria

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Background: In Chronic Obstructive Pulmonary diseases (COPD) pathogenesis oxidative stress plays an important role. Hypoxia-Inducible factor (HIF-1α) is a dimeric protein complex which Functions as a master transcriptional regulator of the adaptive response to hypoxiaand is a risk factor that increases when oxidative stress triggers. The role ofHIF-1αin COPD due to smoking is lacking. Aim: This study aims to evaluate the role of HIF-1α in smoker COPD patients comparing its association with diseases severity. Method: In this cross-sectional study, we recruited 87 subjects, 57 were smokers with COPD,15 were smokers without COPD and other 15 were non-smoker healthy controls. The mean age was 54.6± 9.32 (cases 57.08±8.15; controls 50.0± 9.8). There were 62%smokers, 25% non-smokers,7% tobacco chewers and 6% ex-smokers. Enzyme-linked immune sorbent assay (ELISA) method was used for analyzing serum samples wherein HIF-1α was analyzed by Sandwich-ELISA. Results: In smoker COPD patients, a significantly higher HIF-1α level showed positive association with hypoxia, smoking status and severity of disease (p=0.03). The mean value of HIF-1α was not significantly different in smokers without COPD and healthy controls. Conclusion: It is found that HIF-1α level was increased in smoker COPD, but not in smokers without COPD. This suggests that development of COPD drive the HIF-1α pathway and it correlates with the severity of diseases.

Keywords: COPD, chronic obstructive pulmonary diseases, smokers, nonsmokers, hypoxia

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Authors: Abdul Manan, Choudhary Muhammad Ayyub, Rashid Ahmad, Muhammad Adnan Bukhari

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A project was designed to investigate the effect of foliar application of methyl jasmonate (MeJA) on physiological, biochemical and ionic attributes of salinity stressed and normal tomato plants at different stages. Salinity stress at every stage markedly reduced the net photosynthetic rate, stomatal conductance, transpiration rate, water relations parameters, protein contents, total free aminoacids and potassium (K+) contents. While, antioxidant enzymes (peroxidase (POX) and catalase (CAT)), sodium (Na+) contents and proline contents were increased substantially. Foliar application of MeJA ameliorated the drastic effects of salinity regime by recovery of physiological and biochemical attributes by enhanced production of antioxidant enzymes and osmoprotectants. The efficacy of MeJA at very initial stage (15 days after sowing (15 DAS)).proved effective for attenuating the deleterious effects of salinity stress than other stages (15 days after transplanting (15 DAT) and 30 days after transplanting (30 DAT)). To the best of our knowledge, different times of foliar feeding of MeJA was observed first time for amelioration of salinity stress in tomato plants that would be of pivotal significance for scientist to better understand the dynamics of physiological and biochemical processes in tomato.

Keywords: methyl jasmonate, osmoregulation, salinity stress, stress tolerance, tomato

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Authors: Farnaz Yusuf, Naseem A. Gaur

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The increase in environmental concerns, rapid depletion of fossil fuel reserves and intense interest in achieving energy security has led to a global research effort towards developing renewable sources of fuels. Second generation biofuels have attracted more attention recently as the use of lignocellulosic biomass can reduce fossil fuel dependence and is environment-friendly. Xylose is the main pentose and second most abundant sugar after glucose in lignocelluloses. Saccharomyces cerevisiae does not readily uptake and use pentose sugars. For an economically feasible biofuel production, both hexose and pentose sugars must be fermented to ethanol. Therefore, it is important to develop S. cerevisiae host platforms with more efficient xylose utilization. This work aims to construct a xylose fermenting yeast strains with engineered oxido-reductative pathway for xylose metabolism. Engineered strain was further improved by adaptive evolutionary engineering approach. The engineered strain is able to grow on xylose as sole carbon source with the maximum ethanol yield of 0.39g/g xylose and productivity of 0.139g/l/h at 96 hours. The further improvement in strain development involves over expression of pentose phosphate pathway and protein engineering of xylose reductase/xylitol dehydrogenase to change their cofactor specificity in order to reduce xylitol accumulation.

Keywords: biofuel, lignocellulosic biomass, saccharomyces cerevisiae, xylose

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Authors: Rosa Buxeda, Lorenzo Saliceti-Piazza, Rodolfo J. Romañach, Luis Ríos, Sandra L. Maldonado-Ramírez

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Biopharmaceuticals manufacturing is one of the major economic activities worldwide. Ninety-three percent of the workforce in a biomanufacturing environment concentrates in production-related areas. As a result, strategic collaborations between industry and academia are crucial to ensure the availability of knowledgeable workforce needed in an economic region to become competitive in biomanufacturing. In the past decade, our institution has been a key strategic partner with multinational biotechnology companies in supplying science and engineering graduates in the field of industrial biotechnology. Initiatives addressing all levels of the educational pipeline, from K-12 to college to continued education for company employees have been established along a ten-year span. The Amgen BioTalents Program was designed to provide undergraduate science and engineering students with training in biomanufacturing. The areas targeted by this educational program enhance their academic development, since these topics are not part of their traditional science and engineering curricula. The educational curriculum involved the process of producing a biomolecule from the genetic engineering of cells to the production of an especially targeted polypeptide, protein expression and purification, to quality control, and validation. This paper will report and describe the implementation details and outcomes of the first sessions of the program.

Keywords: biomanufacturing curriculum, interdisciplinary learning, workforce development, industry-academia partnering

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Authors: Gargi Ghoshal, Ashay Jain

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Microencapsulation of β-carotene was optimized by complex coacervation technique using casein/gum tragacanth (CAS/GT) coating as a function of pH, initial protein to polysaccharide mixing ratio (Pr:Ps), total biopolymer concentration, core material load, zeta potential, and ionic strength. This study was aimed to understand the influence of experimental parameters on the coacervation kinetics, the coacervate yield, and entrapment efficiency. At a Pr:Ps = 2:1, an optimum pH of complex coacervation was found 4.35, at which the intensity of electrostatic interaction was maximum. At these ratios of coating, the phase separation occurred the fastest and the final coacervate yield and entrapment efficiency was the highest. Varying the Pr: Ps shifted the value of optimum pH. This incident was due to the level of charge compensation of the CAS/GT complexes. Finally, electrostatic interaction and formation of coacervates between CAS and GT were confirmed by Fourier transform infra-red (FTIR) spectra. The size and surface properties of coacervates were studied using scanning electron microscopy (SEM). The resultant formulation (β-carotene loaded microcapsules) was evaluated for in vitro release study and antioxidant activity. Stability of encapsulated β-carotene was also evaluated under three levels of temperature (5, 25 and 40 °C) for 3 months. Encapsulation strongly increased the stability of micronutrients. Our results advocate potential of microcapsules as a novel carrier for the safeguard and sustained release of micronutrient.

Keywords: β-carotene, casein, complex coacervation, controlled release, gum tragacanth, microcapsules

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Authors: Hsin-Lien Lin, Ju-Hui Fu

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Dendritic cells (DCs) are the major professional antigen-presenting cells for the development of optimal T-cell immunity. DCs can be used as pharmacological targets to screen novel biological modifiers for the treatment of harmful immune responses, such as transplantation rejection. Dryopteris crassirhizoma Nakai (Aspiadaceae) is used for traditional herbal medicine in the region of East Asia. The root of this fern plant has been listed for treating inflammatory diseases. Dryocrassin is the tetrameric phlorophenone component derived from Dryopteris. Here, we tested the immunomodulatory potential of dryocrassin on lipopolysaccharide (LPS)-stimulated activation of mouse bone marrow-derived DCs in vitro and in skin allograft transplantation in vivo. Results demonstrated that dryocrassin reduced the secretion of tumor necrosis factor-α, interleukin-6, and interleukin-12p70 by LPS-stimulated DCs. The expression of LPS-induced major histocompatibility complex class II, CD40, and CD86 on DCs was also blocked by dryocrassin. Moreover, LPS-stimulated DC-elicited allogeneic T-cell proliferation was lessened by dryocrassin. In addition, dryocrassin inhibited LPS-induced activation of IϰB kinase, JNK/p38 mitogen-activated protein kinase, as well as the translocation of NF-ϰB. Treatment with dryocrassin obviously diminished 2,4-dinitro-1-fluorobenzene- induced delayed-type hypersensitivity and prolonged skin allograft survival. Dryocrassin may be one of the potent immunosuppressive agents for transplant rejection through the destruction of DC maturation and function.

Keywords: dryocrassin, dendritic cells, immunosuppression, skin allograft

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Authors: Nadia A. Mohamed, Zakaria El-Khayat, Wagdy K. B. Khalil, Mehrez E. El-Naggar

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Drug nephrotoxicity is still a problem for patients who have taken drugs for elongated periods or permanently. Ultrasound-assisted sol−gel method was used to prepare hollow structured poroussilica nanoemulsion loaded with sodium salicylate as a model drug. The work was extended to achieve the target of the current work via investigating the protective role of this nanoemulsion model as anti-inflammatory drug or ginger for its antioxidant effect against cisplatin-induced nephrotoxicity in male albino rats. The results clarify that the nanoemulsion model was synthesized using ultrasonic assisted with small size and well stabilization as proved by TEM and DLS analysis. Additionally, blood urea nitrogen (BUN), Serum creatinine (SC) and Urinary total protein (UTP) were increased, and the level of creatinine clearance (Crcl) was decreased. All those were met with disorders in oxidative stress and downregulation in the expression of the nephrin gene. Also, histopathological changes of the kidney tissue were observed. These changes back to normal by treatment with silica nanoparticles loaded sodium salicylate (Si-Sc-NPs), ginger or both. Conclusions oil/water nanoemulsion of (Si-Sc NPs) and ginger showed a protective and promising preventive strategy against nephrotoxicity due to their antioxidant and anti-inflammatory effects, and that offers a new approach in attenuating drug induced nephrotoxicity.

Keywords: sodium salicylate nanoencapsulation, nephrin mRNA, drug nephrotoxicity, cisplatin, experimental rats

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Authors: Wenjun Zhang, Yunqing Du, Steven W. Cranford, Ming L. Wang

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The beginning of 21st century has witnessed new advancements in the design and use of new materials for biosensing applications, from nano to macro, protein to tissue. Traditional analytical methods lack a complete toolset to describe the complexities introduced by living systems, pathological relations, discrete hierarchical materials, cross-phase interactions, and structure-property dependencies. Materiomics – via systematic molecular dynamics (MD) simulation – can provide structure-process-property relations by using a materials science approach linking mechanisms across scales and enables oriented biosensor design. With this approach, DNA biosensors can be utilized to detect disease biomarkers present in individuals’ breath such as acetone for diabetes. Our wireless sensor array based on single-stranded DNA (ssDNA)-decorated single-walled carbon nanotubes (SWNT) has successfully detected trace amount of various chemicals in vapor differentiated by pattern recognition. Here, we present how MD simulation can revolutionize the way of design and screening of DNA aptamers for targeting biomarkers related to oral diseases and oral health monitoring. It demonstrates great potential to be utilized to build a library of DNDA sequences for reliable detection of several biomarkers of one specific disease, and as well provides a new methodology of creating, designing, and applying of biosensors.

Keywords: biosensor, DNA, biomarker, molecular dynamics simulation

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Authors: Essam A. Makky, Siti H. Mohd Rasdi, J. B. Al-Dabbagh, G. F. Najmuldeen

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The synthesis of silver nano particles (SNPs) extensively studied by using chemical and physical methods. Here, the biological methods were used and give benefits in research field in the aspect of very low cost (from waste to wealth) and safe time as well. The study aims to isolate and exploit the microbial power in the production of industrially important by-products in nano-size with high economic value, to extract highly valuable materials from hazardous waste, to quantify nano particle size, and characterization of SNPs by X-Ray Diffraction (XRD) analysis. Disposal X-ray films were used as substrate because it consumes about 1000 tons of total silver chemically produced worldwide annually. This silver is being wasted when these films are used and disposed. Different bacterial isolates were obtained from various sources. Silver was extracted as nano particles by microbial power degradation from disposal X-ray film as the sole carbon source for ten days incubation period in darkness. The protein content was done and all the samples were analyzed using XRD, to characterize of silver (Ag) nano particles size in the form of silver nitrite. Bacterial isolates CL4C showed the average size of SNPs about 19.53 nm, GL7 showed average size about 52.35 nm and JF Outer 2A (PDA) showed 13.52 nm. All bacterial isolates partially identified using Gram’s reaction and the results obtained exhibited that belonging to Bacillus sp.

Keywords: nanotechnology, bioremediation, disposal X-ray film, nanoparticle, waste, XRD

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Authors: Sepideh Hassanpour Khodaei

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Background/Objectives: The purpose of this study is to evaluate the effect of mummy substances on two issues of proliferation and production of matrix protein synthesis in wound healing. Methods: The methodology used for this aim involves isolating mesenchymal stem cells and human fibroblasts procured at Pastor Institute, Iran. The cells were treated with mummy substances separately and co-cultured between ASCs and WJSCs, and fibroblasts. Proliferation was assessed by Ki67 method in monolayer conditions. Synthesis of components of extracellular matrix (ECM) such as collagen type I, type III, and fibronectin 1 (FN1) was determined by qPCR. Results: The effects of adipocyte stem cells (ASCs), Wharton Jelly Stem Cells (WJSCs), and Mummy material on fibroblast proliferation and migration were evaluated. The present finding underlined the importance of Mummy material, ASCs, and WJSCs in the proliferation and migration of fibroblast cells. Furthermore, the expression of collagen I, III, and FN1 was increased in the presence of the above material and cells. Conclusion: This study presented an effective in vitro method for the healing process. Hence, the prospect of utilizing Mummy material and stem cell-based therapies in wound healing as a therapeutic approach is promising.

Keywords: mummy material, wound healing, adipose tissue, Wharton’s jelly

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Authors: Chowon Kim, Yoo-Kyung Kim, Kyeong Yeon Park, Hyun-Shik Lee

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Perfluoroalkyl compounds (PFCs) are globally distributed synthetic compounds that are known to adversely affect human health. Developmental toxicity assessment of PFCs is important to facilitate the evaluation of their environmental impact. In the present study, we assessed the developmental toxicity and teratogenicity of PFCs with different numbers of carbon atoms on Xenopus embryogenesis. An initial frog embryo teratogenicity assay-Xenopus (FETAX) assay was performed that identified perfluorohexanoic (PFHxA) and perfluoroheptanoic (PFHpA) acids as potential teratogens and developmental toxicants. The mechanism underlying this teratogenicity was also investigated by measuring the expression of tissue-specific biomarkers such as phosphotyrosine‑binding protein, xPTB (liver); NKX2.5 (heart); and Cyl18 (intestine). Whole‑mount in situ hybridization, reverse transcriptase‑polymerase chain reaction (RT-PCR), and histologic analyses detected severe defects in the liver and heart following exposure to PFHxA or PFHpA. In addition, immunoblotting revealed that PFHpA significantly increased the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), while PFHxA slightly increased these, as compared with the control. These results suggest that PFHxA and PFHpA are developmental toxicants and teratogens, with PFHpA producing more severe effects on liver and heart development through the induction of ERK and JNK phosphorylation.

Keywords: PFCs, ERK, JNK, xenopus

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Authors: Amel A. Refaie, Amal Ramadan, Abdel-Tawab H. Mossa

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This study was designed to evaluate the adverse effects of sub-chronic exposure to the fipronil on the liver of female rats at a dose equal to 400 mg /kg (1/10LD50) in drinking water and the protective role of fish oil at concentration 117.6 mg/Kg b.wt via oral routes daily for 28 days. Fipronil treatment caused a decrease in body weight gain and increase in relative liver weight. Fipronil induced a significant increase in the liver biomarkers enzymes such as alanine aminotransferases (ALT), aspartate aminotransferases (AST), alkaline phosphatase (ALP) and levels of total protein while fipronil caused a significant decrease in butyryl cholinesterase activity in FPN-treated rats. Oxidative stress biomarkers such as superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) were significantly decreased in liver tissue, while lipid peroxidation (LPO) was significantly increased in fipronil treating rats in a dose-dependent manner. FPN caused histopathological alterations in liver of female rats. From our results, it can be reported that FPN induced lipid peroxidation, oxidative stress, liver injury in female rats and fish oil used to protect animals against the adverse effect of pesticide exposure. These pathophysiological alterations in liver tissues could be due to the toxic effect of fipronil that associated with a generation of free radicals.

Keywords: fipronil (FPN), fish oil, hepatotoxicity, transaminases, antioxidant enzymes, female rats

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Authors: Sagri Sharma

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Analysis of diseases integrating multi-factors increases the complexity of the problem and therefore, development of frameworks for the analysis of diseases is an issue that is currently a topic of intense research. Due to the inter-dependence of the various parameters, the use of traditional methodologies has not been very effective. Consequently, newer methodologies are being sought to deal with the problem. Supervised Learning Algorithms are commonly used for performing the prediction on previously unseen data. These algorithms are commonly used for applications in fields ranging from image analysis to protein structure and function prediction and they get trained using a known dataset to come up with a predictor model that generates reasonable predictions for the response to new data. Gene expression profiles generated by DNA analysis experiments can be quite complex since these experiments can involve hypotheses involving entire genomes. The application of well-known machine learning algorithm - Support Vector Machine - to analyze the expression levels of thousands of genes simultaneously in a timely, automated and cost effective way is thus used. The objectives to undertake the presented work are development of a methodology to identify genes relevant to Hepatocellular Carcinoma (HCC) from gene expression dataset utilizing supervised learning algorithms and statistical evaluations along with development of a predictive framework that can perform classification tasks on new, unseen data.

Keywords: artificial intelligence, biomarker, gene expression datasets, hepatocellular carcinoma, machine learning, supervised learning algorithms, support vector machine

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Authors: Kelsi Hagerty, Ami Rosen, Aaliyah Heyward, Nadia Ali, Emily Brown, Erin Demo, Yue Guan, Modele Ogunniyi, Brianna McDaniels, Alanna Morris, Kunal Bhatt

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Hereditary transthyretin amyloidosis (hATTR) is a progressive, multi-systemic, and life-threatening disease caused by a disruption in the TTR protein that delivers thyroxine and retinol to the liver. This disruption causes the protein to misfold into amyloid fibrils, leading to the accumulation of the amyloid fibrils in the heart, nerves, and GI tract. Over 130 variants in the TTR gene are known to cause hATTR. The Val122Ile variant is the most common in the United States and is seen almost exclusively in people of African descent. TTR variants are inherited in an autosomal dominant fashion and have incomplete penetrance and variable expressivity. Individuals with hATTR may exhibit symptoms from as early as 30 years to as late as 80 years of age. hATTR is characterized by a wide range of clinical symptoms such as cardiomyopathy, neuropathy, carpal tunnel syndrome, and GI complications. Without treatment, hATTR leads to progressive disease and can ultimately lead to heart failure. hATTR disproportionately affects individuals of African descent; the estimated prevalence of hATTR among Black individuals in the US is 3.4%. Unfortunately, hATTR is often underdiagnosed and misdiagnosed because many symptoms of the disease overlap with other cardiac conditions. Due to the progressive nature of the disease, multi-systemic manifestations that can lead to a shortened lifespan, and the availability of free genetic testing and promising FDA-approved therapies that enhance treatability, early identification of individuals with a pathogenic hATTR variant is important, as this can significantly impact medical management for patients and their relatives. Furthermore, recent literature suggests that TTR genetic testing should be performed in all patients with suspicion of TTR-related cardiomyopathy, regardless of age, and that follow-up with genetic counseling services is recommended. Relatives of patients with hATTR benefit from genetic testing because testing can identify carriers early and allow relatives to receive regular screening and management. Despite the striking prevalence of hATTR among Black individuals, hATTR remains underdiagnosed in this patient population, and germline genetic testing for hATTR in Black individuals seems to be underrepresented, though the reasons for this have not yet been brought to light. Historically, Black patients experience a number of barriers to seeking healthcare that has been hypothesized to perpetuate the underdiagnosis of hATTR, such as lack of access and mistrust of healthcare professionals. Prior research has described a myriad of factors that shape an individual’s decision about whether to pursue presymptomatic genetic testing for a familial pathogenic variant, such as family closeness and communication, family dynamics, and a desire to inform other family members about potential health risks. This study explores these factors through 10 in-depth interviews with patients with hATTR about what factors may be contributing to the underdiagnosis of hATTR in the Black population. Participants were selected from the Emory University Amyloidosis clinic based on having a molecular diagnosis of hATTR. Interviews were recorded and transcribed verbatim, then coded using MAXQDA software. Thematic analysis was completed to draw commonalities between participants. Upon preliminary analysis, several themes have emerged. Barriers identified include i) Misdiagnosis and a prolonged diagnostic odyssey, ii) Family communication and dynamics surrounding health issues, iii) Perceptions of healthcare and one’s own health risks, and iv) The need for more intimate provider-patient relationships and communication. Overall, this study gleaned valuable insight from members of the Black community about possible factors contributing to the underdiagnosis of hATTR, as well as potential solutions to go about resolving this issue.

Keywords: cardiac amyloidosis, heart failure, TTR, genetic testing

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Authors: Arianne V. Julian, Michael R. Umagat, Renato G. Reyes

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Pleurotus ostreatus, which is one of the most widely cultivated mushrooms, is an excellent source of protein and other minerals but inherently contains low calcium level. Calcium plays several vital functions in human health; therefore, adequate daily intake is necessary. Supplementation of growth substrate is a significant approach in mushroom production to improve nutritional content and yield. This study focused on the influence of varying concentrations of Ca supplementation derived from natural sources including agricultural lime, eggshell and oyster shell in rice straw-based formulation for the production of P. ostreatus. The effect of Ca supplementation on the total yield and Ca content were obtained. Results revealed that these natural sources increased both the yield and Ca of P. ostreatus. Mushroom grown in substrate with 8-10% agricultural lime and 6% eggshell powder produced the highest yields while using oyster shell powder did not vary with the control. Meanwhile, substrate supplementation using agricultural lime and eggshell powder in all concentrations have increased Ca in fruiting bodies. However, Ca was not absorbed in the oyster shell powder-supplemented substrate. These findings imply the potential of agricultural lime and eggshell powder in the production of Ca-enriched mushrooms resulting in higher yield.

Keywords: calcium fortification, mushroom production, natural sources, Pleurotus ostreatus

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Authors: Nirupama Dixit, Anyaa Mittal, Neeru Sood

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Alzheimer’s disease (AD) is a progressive form of dementia, contributing to up to 70% of cases, mostly observed in elderly but is not restricted to old age. The pathophysiology of the disease is characterized by specific pathological changes in brain. The changes (i.e. accumulation of metal ions in brain, formation of extracellular β-amyloid (Aβ) peptide aggregates and tangle of hyper phosphorylated Tau protein inside neurons) damage the neuronal connections irreversibly. The current issues in improvement of life quality of Alzheimer's patient lies in the fact that the diagnosis is made at a late stage of the disease and the medications do not treat the basic causes of Alzheimer's. The targeted delivery of drug through the blood brain barrier (BBB) poses several limitations via traditional approaches for treatment. To overcome these drug delivery limitation, nanoparticles provide a promising solution. This review focuses on current strategies for efficient targeted drug delivery using nanoparticles and improving the quality of therapy provided to the patient. Nanoparticles can be used to encapsulate drug (which is generally hydrophobic) to ensure its passage to brain; they can be conjugated to metal ion chelators to reduce the metal load in neural tissue thus lowering the harmful effects of oxidative damage; can be conjugated with drug and monoclonal antibodies against BBB endogenous receptors. Finally this review covers how the nanoparticles can play a role in diagnosing the disease.

Keywords: Alzheimer's disease, β-amyloid plaques, blood brain barrier, metal chelators, nanoparticles

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Authors: Hanan, S. Siam, Safaa A. Mahmoud, A. S. Taalab

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A pot experiment was conducted in greenhouse of NRC, Dokki, Cairo, Egypt to study the response of wheat plants to different levels of superphosphate at (60kg P2O5 or 30 kg P2O5) with or without potassium silicate or chicken compost (2.5 ton/fed.) on growth yield and nutrients status especially, and phosphorus and silica availability. Data reveal that the addition either chicken or compost increased significantly affected on all the growth and yield parameters as well as nutrients status and protein of the different parts of wheat plants if compared with control (60kg P2O5 or 30 kg P2O5). Data also reveal that the highest mean values were obtained when potassium silicate with was added to 60 kg P2O5, while the lowest values of the previous parameters were obtained when 30 kg P2O5 alone was added to plants. Furthermore, data indicated that the highest mean values of all mentioned parameters were obtained when chicken compost was applied with any rate of P as compared with silica addition at the same rates of P. According to the results, the highest values of all mentioned parameters were obtained when addition of chicken compost and potassium silicate including the high rate of P at (60 kg P2O5) while the lowest values of the previous parameters were obtained when plants received of phosphorus (30 kg P2O5) alone.

Keywords: wheat, yield, chicken compost, potassium, phosphorus, silicate, nutrients status

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Authors: Cahit Demetgul, Sahin Bayraktar, Neslihan Beyazit

Abstract:

Metalloenzymes are enzyme proteins containing metal ions, which are directly bound to the protein or to enzyme-bound nonprotein components. One of the major metalloenzymes that play a key role in oxidation reactions is catechol oxidase, which shows catecholase activity i.e. oxidation of a broad range of catechols to quinones through the four-electron reduction of molecular oxygen to water. Studies on the model compounds mimicking the catecholase activity are very useful and promising for the development of new, more efficient bioinspired catalysts, for in vitro oxidation reactions. In this study, a new tetradentate asymmetrical Schiff-base and its Cu(II) complex were synthesized by condensation of 4-nitro-1,2-phenylenediamine with 6-formyl-7-hydroxy-5-methoxy-2-methylbenzopyran-4-one and by using an appropriate Cu(II) salt, respectively. The prepared compounds were characterized by elemental analysis, FT-IR, NMR, UV-Vis and magnetic susceptibility. The catecholase-mimicking activity of the new Schiff Base Cu(II) complex was performed for the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC) in methanol at 25 °C, where the electronic spectra were recorded at different time intervals. The yield of the quinone (3,5-DTBQ) was determined from the measured absorbance at 400 nm of the resulting solution. The compatibility of catalytic reaction with Michaelis-Menten kinetics was also investigated. In conclusion, we have found that our new Schiff Base Cu(II) complex presents a significant capacity to catalyze the oxidation reaction of the catechol to o-quinone.

Keywords: catecholase activity, Michaelis-Menten kinetics, Schiff base, transition metals

Procedia PDF Downloads 293

Authors: Ritesh K. Shukla

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Degradation of biological samples in terms of macromolecules (DNA, RNA, and protein) are the major challenges in the forensic investigation which misleads the result interpretation. Currently, there are no precise methods available to circumvent this problem. Therefore, at the preliminary level, some methods are urgently needed to solve this issue. In this order, Comet assay is one of the most versatile, rapid and sensitive molecular biology technique to assess the DNA degradation. This technique helps to assess DNA degradation even at very low amount of sample. Moreover, the expedient part of this method does not require any additional process of DNA extraction and isolation during DNA degradation assessment. Samples directly embedded on agarose pre-coated microscopic slide and electrophoresis perform on the same slide after lysis step. After electrophoresis microscopic slide stained by DNA binding dye and observed under fluorescent microscope equipped with Komet software. With the help of this technique extent of DNA degradation can be assessed which can help to screen the sample before DNA fingerprinting, whether it is appropriate for DNA analysis or not. This technique not only helps to assess degradation of DNA but many other challenges in forensic investigation such as time since deposition estimation of biological fluids, repair of genetic material from degraded biological sample and early time since death estimation could also be resolved. With the help of this study, an attempt was made to explore the application of well-known molecular biology technique that is Comet assay in the field of forensic science. This assay will open avenue in the field of forensic research and development.

Keywords: comet assay, DNA degradation, forensic, molecular biology

Procedia PDF Downloads 135

Authors: Latifa Hamoudi-Belarbi, L'Hadi Nouri, Khaled Belkacemi

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The effect of three methods of drying (traditional drying, freeze-drying and spray-drying) on the survival of concentrated cultures of Geotrichum fragrans and Wickerhamomyces anomalus was studied. The survival of yeast cultures was initially compared immediately after freeze-drying using HES 12%(w/v)+Sucrose 7% (w/v) as protectant, traditional drying in dry rice cakes and finally spray-drying with whey proteins. The survival of G. fragrans and W. anomalus was studied during 4 months of storage at 4°C and 25°C, in the darkness, under vacuum and at 0% relative humidity. The results demonstrated that high survival was obtained using traditional method of preservation in rice cakes (60% for G. fragrans and 65% for W. anomalus) and freeze-drying in (68% for G. fragrans and 74% for W. anomalus). However, poor survival was obtained by spray-drying method in whey protein with 20% for G. fragrans and 29% for W. anomalus. During storage at 25°C, yeast cultures of G. fragrans and W. anomalus preserved by traditional and freeze-drying methods showed no significant loss of viable cells up to 3 months of storage. Spray-dried yeast cultures had the greatest loss of viable count during the 4 months of storage at 25°C. During storage at 4°C, preservation of yeasts cultures using traditional method of preservation provided better survival than freeze-drying. This study demonstrated the effectiveness of the traditional method to preserve yeasts cultures compared to the high cost methods like freeze-drying and spray-drying.

Keywords: freeze-drying, traditional drying, spray drying, yeasts

Procedia PDF Downloads 466

Authors: Fariha Akhter Chowdhury, Mohammad Nurul Islam, Anamika Saha, Sabrina Mahboob, Abu Syed Md. Mosaddek, Md. Omar Faruque, Most. Fahmida Begum, Rajib Bhattacharjee

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Urinary tract infection (UTI) is one of the most common infectious diseases in Bangladesh where Escherichia coli is the prevalent organism and responsible for most of the infections. Lipopolysaccharide (LPS) is known to act as a major virulence factor of E. coli. The present study aimed to purify, extract and visualize LPS of E. coli clinical isolates from urine samples of patients with UTI. The E. coli strain was isolated from the urine samples of 10 patients with UTI and then the antibiotic sensitivity pattern of the isolates was determined. The purification of LPS was carried out using the hot aqueous-phenol method and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, which was directly stained using the modified silver staining method and Coomassie blue. The silver-stained gel demonstrated both smooth and rough type LPS by showing trail-like band patterns with the presence and lacking O-antigen region, respectively. Coomassie blue staining showed no band assuring the absence of any contaminating protein. Our successful extraction of purified LPS from E. coli isolates of UTI patients’ urine samples can be an important step to understand the UTI disease conditions.

Keywords: Escherichia coli, electrophoresis, polyacrylamide gel, silver staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

Procedia PDF Downloads 360

Authors: Hamidreza Miri, Farahnaz Momtazi

Abstract:

Wheat is one of the most important crops in Fars province, and because of water shortage, there is a great emphasis on its water use efficiency in the production field. A field experiment was conducted in 2021 and 2022 in order to evaluate wheat yield and its components in raised planting system in Arsanjan, Fars province. The experiment was conducted as a split plot with three irrigation treatments (irrigation equal to evapotranspiration, 80% of evapotranspiration irrigation (moderate drought stress), and 60% of evapotranspiration irrigation (severe drought stress)) as the main plot and three planting methods (conventional flat planting, 60 cm raised bed planting and 120 cm raised bed planting) as a subplot. The results indicated that drought stress significantly decreased traits such as plant height, grain yield, ear number, seed number, and biological yield while increasing seed protein. Raised bed planting significantly increased the traits in comparison with conventional flat planting. So that plating with a 120 cm raised bed increased grain yield by 22.1% and 25.9% in the first and second years, respectively. This increase was 17% for biological, 75 for ear number, and 21% for seed number. Planting in raised bed system reduced the adverse effect of drought stress on wheat traits. In conclusion, based on the observed results planting in raised bed system can be adopted as an appropriate planting pattern for improving yield and water productivity in experimental regions and similar climates.

Keywords: wheat, raised bed planting, drought stress, yield, water use

Procedia PDF Downloads 46