Search results for: fusion proteins

Authors: J. D. Cabral, E. Murray, P. Turner, E. Hewitt, A. Ali, M. McConnell

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Worldwide demand for organ replacement and tissue regeneration is progressively increasing. Three-dimensional (3D) bioprinting, where a physical construct is produced using computer-aided design, is a promising tool to advance the tissue engineering and regenerative medicine fields. In this paper we describe two different approaches to developing 3D bioprinted constructs for use in tissue regeneration. Bioink development is critical in achieving the 3D biofabrication of functional, regenerative tissues. Hydrogels, cross-linked macromolecules that absorb large amounts of water, have received widespread interest as bioinks due to their relevant soft tissue mechanics, biocompatibility, and tunability. In turn, not only is bioink optimisation crucial, but the creation of vascularized tissues remains a key challenge for the successful fabrication of thicker, more clinically relevant bioengineered tissues. Among the various methodologies, cell-laden hydrogels are regarded as a favorable approach; and when combined with novel core/shell 3D bioprinting technology, an innovative strategy towards creating new vessel-like structures. In this work, we investigate this cell-based approach by using human umbilical endothelial cells (HUVECs) entrapped in a viscoelastic chitosan/dextran (CD)-based core hydrogel, printed simulataneously along with a gelatin methacrylate (GelMA) shell. We have expanded beyond our previously reported FDA approved, commercialised, post-surgical CD hydrogel, Chitogel®, by functionalizing it with cell adhesion and proteolytic peptides in order to promote bone marrow-derived mesenchymal stem cell (immortalized BMSC cell line, hTERT) and HUVECs growth. The biocompatibility and biodegradability of these cell lines in a 3D bioprinted construct is demonstrated. Our studies show that particular peptide combinations crosslinked within the CD hydrogel was found to increase in vitro growth of BMSCs and HUVECs by more than two-fold. These gels were then used as a core bioink combined with the more mechanically robust, UV irradiated GelMA shell bioink, to create 3D regenerative, vessel-like scaffolds with high print fidelity. As well, microporous MEW scaffolds made from milk proteins blended with PCL were found to show promising bioactivity, exhibiting a significant increase in keratinocyte (HaCaTs) and fibroblast (normal human dermal fibroblasts, NhDFs) cell migration and proliferation when compared to PCL only scaffolds. In conclusion, our studies indicate that a peptide functionalized CD hydrogel bioink reinforced with a GelMA shell is biocompatible, biodegradable, and an appropriate cell delivery vehicle in the creation of regenerative 3D constructs. In addition, a novel 3D printing technique, melt electrowriting (MEW), which allows fabrication of micrometer fibre meshes, was used to 3D print polycaprolactone (PCL) and bioactive milk protein, lactorferrin (LF) and whey protein (WP), blended scaffolds for potential skin regeneration applications. MEW milk protein/PCL scaffolds exhibited high porosity characteristics, low overall biodegradation, and rapid protein release. Human fibroblasts and keratinocyte cells were seeded on to the scaffolds. Scaffolds containing high concentrations of LF and combined proteins (LF+WP) showed improved cell viability over time as compared to PCL only scaffolds. This research highlights two scaffolds made using two different 3D printing techniques using a combination of both natural and synthetic biomaterial components in order to create regenerative constructs as potential chronic wound treatments.

Keywords: biomaterials, hydrogels, regenerative medicine, 3D bioprinting

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Authors: Michella Alnajjar, Frederic Christien, Krzysztof Wolski, Cedric Bosch

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Additive manufacturing (AM) has gained more interest in the past few years because it allows 3D parts often having a complex geometry to be directly fabricated, layer by layer according to a CAD model. One of the AM techniques is the selective laser melting (SLM) which is based on powder bed fusion. In this work, the corrosion resistance of 17-4 PH steel obtained by SLM is investigated. Wrought 17-4 PH steel is a martensitic precipitation hardenable stainless steel. It is widely used in a variety of applications such as aerospace, medical and food industries, due to its high strength and relatively good corrosion resistance. However, the combined findings of X-Ray diffraction and electron backscatter diffraction (EBSD) proved that SLM-ed 17-4 PH steel has a fully ferritic microstructure, more specifically δ ferrite. The microstructure consists of coarse ferritic grains elongated along the build direction, with a pronounced solidification crystallographic texture. These results were associated with the high cooling and heating rates experienced throughout the SLM process (10⁵-10⁶ K/s) that suppressed the austenite formation and produced a 'by-passing' phenomenon of this phase during the numerous thermal cycles. Furthermore, EDS measurements revealed a uniform distribution of elements without any dendritic structure. The extremely high cooling kinetics induced a diffusionless solidification, resulting in a homogeneous elemental composition. Consequently, the corrosion properties of this steel are altered from that of conventional ones. By using electrochemical means, it was found that SLM-ed 17-4 PH is more resistant to general corrosion than the wrought steel. However, the SLM-ed material exhibits metastable pitting due to its high porosity density. In addition, the hydrogen embrittlement of SLM-ed 17-4 PH steel is investigated, and a correlation between its behavior and the observed microstructure is made.

Keywords: corrosion resistance, 17-4 PH stainless steel, selective laser melting, hydrogen embrittlement

Procedia PDF Downloads 141

Authors: Seung-Hwa Baek, In-Jung Nam, Sang-Han Lee

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The development of anti-melanogenic agents is important for the prevention of serious esthetic problem like a melasma, freckle, age spots, and chloasma. The aim of this study was to investigate the anti-melanogenic effect of sesamol, an active lignan isolated from sesame seed, by mushroom and cellular tyrosinase assay, melanin content and the analysis of melanogensis-related mRNA expressions in melana cells. Sesamol showed strong inhibitory activity against the mushroom tyrosinase in a dose-dependent manner. Intracellular tyrosinase inhibition activity was also confirmed by zymography. At a concentration of 50 μM, sesamol inhibited melanin production in melan-a cells with no cytoxicity while those of phenylthiourea (PTU) as a positive control were the same condition. Sesamol significantly inhibited the expression of melanogensis-related genes, such as tyrosinase, tyrosinase-related protein-1 (TRP-1), dopachrome tautomerase (Dct), microphthalmia-associated transcription factor (MITF) and melanocortin 1 receptor (MC1R). These findings indicate that sesamol could reduce melanin biosynthesis via the downregulation of tyrosinase activity and melanin production via subsequent gene expression of melanogenesis-related proteins. Together, these results suggest that the sesamol have strong potential in inhibiting melanin biosynthesis, in that the substance may be used as a new skin-whitening agent of cosmetic materials.

Keywords: sesamol, sesame seed, melanin biosynthesis, melanogenesis-related gene, skin-whitening agent

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Authors: Tunca Gul Altuntas, Aziz Baydar, Cemre Acar, Sezen Yılmaz, Tulay Coban

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Free radicals, which are generated in many bioorganic redox processes, play a role in the pathogenesis of several diseases including cancer, arthritis, hemorrhagic shock, inflammatory, cardiovascular, neurodegenerative diseases and age-related degenerative brain diseases. Exposures of normal cell to free radical damages several structures, oxidizes nucleic acids, proteins, lipids, or DNA. Compounds interfere with the action of reactive oxygen species might be useful in prevention and treatment of these pathologies. A series of indole compounds containing piperazine ring were synthesized. Coupling of indole-2-carboxylic acid with monosubstituted piperazines was accomplished with 1,1’-carbonyldiimidazole (CDI) in a good yield. The structures of prepared compounds were verified in good agreement with their 1H NMR (nuclear magnetic resonance), MS (mass spectrophotometry), and IR (infrared spectrophotometry) characteristics. In this work, all synthetized indole derivatives were screened in vitro for their antioxidative potential against vitamin E (α-tocopherol) using different antioxidant assays such as superoxide anion formation, lipid peroxidation levels in rat liver, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) stable radical scavenging activity. The synthesized compounds showed various levels of inhibition compared to vitamin E. This may give promising results for the development of new antioxidant agents.

Keywords: antioxidant, indoles, piperazines, reactive oxygen species

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Authors: Sakina Fatima, Joseph-Patrick W. E. Clarke, Patricia A. Thibault, Subha Kalyaanamoorthy, Michael Levin, Aravindhan Ganesan

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Heteronuclear ribonucleoprotein (hnRNPA1 or A1) is associated with the pathology of different diseases, including neurological disorders and cancers. In particular, the aggregation and dysfunction of A1 have been identified as a critical driver for neurodegeneration (NDG) in Multiple Sclerosis (MS). Structurally, A1 includes a low-complexity domain (LCD) and two RNA-recognition motifs (RRMs), and their interdomain coordination may play a crucial role in A1 aggregation. Previous studies propose that RNA-inhibitors or nucleoside analogs that bind to RRMs can potentially prevent A1 self-association. Therefore, molecular-level understanding of the structures, dynamics, and nucleotide interactions with A1 RRMs can be useful for developing therapeutics for NDG in MS. In this work, a combination of computational modelling and biochemical experiments were employed to analyze a set of RNA-A1 RRM complexes. Initially, the atomistic models of RNA-RRM complexes were constructed by modifying known crystal structures (e.g., PDBs: 4YOE and 5MPG), and through molecular docking calculations. The complexes were optimized using molecular dynamics simulations (200-400 ns), and their binding free energies were computed. The binding affinities of the selected complexes were validated using a thermal shift assay. Further, the most important molecular interactions that contributed to the overall stability of the RNA-A1 RRM complexes were deduced. The results highlight that adenine and guanine are the most suitable nucleotides for high-affinity binding with A1. These insights will be useful in the rational design of nucleotide-analogs for targeting A1 RRMs.

Keywords: hnRNPA1, molecular docking, molecular dynamics, RNA-binding proteins

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Authors: Umadevi B., Dinesh P. A., Indira. R., Vinay C. V.

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A mathematical model is developed to study the simultaneous effects of particle drag and slip parameter on the velocity as well as rate of flow in an annular cross sectional region bounded by two eccentric cylinders. In physiological flows this phenomena can be observed in an eccentric catheterized artery with inner cylinder wall is impermeable and outer cylinder wall is permeable. Blood is a heterogeneous fluid having liquid phase consisting of plasma in which a solid phase of suspended cells and proteins. Arterial wall gets damaged due to aging and lipid molecules get deposited between damaged tissue cells. Blood flow increases towards the damaged tissues in the artery. In this investigation blood is modeled as two phase fluid as one is a fluid phase and the other is particulate phase. The velocity of the fluid phase and rate of flow are obtained by transforming eccentric annulus to concentric annulus with the conformal mapping. The formulated governing equations are analytically solved for the velocity and rate of flow. The numerical investigations are carried out by varying eccentricity parameter, slip parameter and drag parameter. Enhancement of slip parameter signifies loss of fluid then the velocity and rate of flow will be decreased. As particulate drag parameter increases then the velocity as well as rate flow decreases. Eccentricity facilitates transport of more fluid then the velocity and rate of flow increases.

Keywords: catheter, slip parameter, drag parameter, eccentricity

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Authors: Gordon James Knowles

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Historically the Chinatown district in Sandwich Islands has been plagued with the traditional vice crimes of illegal drugs, gambling, and prostitution since the early 1800s. However, a new form of psychologically addictive arcade style table gambling machines has become the dominant form of illegal revenue made in Honolulu, Hawaii. This study attempts to document the drive, desire, or will to play and wager with arcade style video gaming and understand the role of illegal game rooms in facilitating pathological gambling addiction. Indicators of police corruption by Chinese organized crime syndicates related to protection rackets, bribery, and pay-offs were revealed. Information fusion from a police science and sociological intelligence perspective indicates insurgent warfare is being waged on the streets of Honolulu by the People’s Republic of China. This state-sponsored communist terrorism in the Hawaiian Islands used “contactless” irregular warfare entailing: (1) the deployment of psychologically addictive gambling machines, (2) the distribution of the physically addictive fentanyl drug as a lethal chemical weapon, and (3) psychological warfare by circulating pro-China anti-American propaganda newspapers targeted at the small island populace.

Keywords: Chinese and Polynesian organized crime, china daily newspaper, electronic arcade style table games, gaming technology addiction, illegal compulsive gambling, and police intelligence

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Authors: Yamani Amal, Lazaae Jamila, Elachouri Mostafa

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Plants and their active agents – especially polyphenols – may have a principal role in the treatment of diseases that result from the defect of physiological antioxidant mechanisms. Bryonia dioica is well known in Moroccan traditional medicine for alleviatin pain and traiting many diseases. We have focused on plant belonging to Cucurbitaceae Family from around the world to understand their therapeutic uses and their potential antioxidant activities Although several biological activities and Chemical composition of Bryonia dioica are well characterized, no direct, in vitro study, of this natural product examined the antioxydant effect of the extract from the roots of Bryonia dioica. The aim of this study was to determine in vitro antioxidant activity of the B.dioica root, using antioxidant analysis methods based on determination of Hydroxyradical Scavenging, 1,1-diphenyl-2-picrylhydrazine (DPPH) radical scavenging, Hydrogenperoxide Scavenging and Nitric Oxide Scavenging. In this study, it was demonstrated, that, B. dioica root extract showed excellent antioxidant properties. This investigation showed that the roots of this plant contain potent natural scavengers R. It may represent an interesting source of antioxidant phenolics that may favour the extension of their cultivation as new source of natural antioxidants in addition to containing high quality proteins for human or animal nutrition. Therefore, there is need for all stakeholders on the Morocco to strive towards taking advantage of our enormous biodiversity resources to free our people from diseases, abject poverty and stagnation.

Keywords: Morocco, bryoniadioica, in vitro, antioxydant

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Authors: Jeong Woo Kang, MiYoung Baek, Ki-Suk Kim, Kwang-Jick Lee, ByungJae So

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Introduction: A fast, easy and sensitive detection method was developed and validated by liquid chromatography tandem mass spectrometry for the determination of marbofloxacin in pig plasma which was further applied to study the pharmacokinetics of marbofloxacin. Materials and Methods: The plasma sample (500 μL) was mixed with 1.5 ml of 0.1% formic acid in MeCN to precipitate plasma proteins. After shaking for 20 min, The mixture was centrifuged at 5,000 × g for 30 min. It was dried under a nitrogen flow at 50℃. 500 μL aliquot of the sample was injected into the LC-MS/MS system. Chromatographic analysis was carried out mobile phase gradient consisting 0.1% formic acid in D.W. (A) and 0.1% formic acid in MeCN (B) with C18 reverse phase column. Mass spectrometry was performed using the positive ion mode and the selected ion monitoring (MRM). Results and Conclusions: The method validation was performed in the sample matrix. Good linearities (R2>0.999) were observed and the quantified average recoveries of marbofloxacin were 87 - 92% at level of 10 ng g-1 -100 ng g-1. The percent of coefficient of variation (CV) for the described method was less than 10 % over the range of concentrations studied. The limits of detection (LOD) and quantification (LOQ) were 2 and 5 ng g-1, respectively. This method has also been applied successfully to pharmacokinetic analysis of marbofloxacin after intravenous (IV), intramuscular (IM) and oral administration (PO). The mean peak plasma concentration (Cmax) was 2,597 ng g-1at 0.25 h, 2,587 ng g-1at 0.44 h and 2,355 ng g-1at 1.58 h for IV, IM and PO, respectively. The area under the plasma concentration-time curve (AUC0–t) was 24.8, 29.0 and 25.2 h μg/mL for IV, IM and PO, respectively. The elimination half-life (T1/2) was 8.6, 13.1 and 9.5 for IV, IM and PO, respectively. Bioavailability (F) of the marbofloxacin in pig was 117 and 101 % for IM and PO, respectively. Based on these result, marbofloxacin does not have any obstacles as therapeutics to develop the oral formulations such as tablets and capsules.

Keywords: marbofloxacin, LC-MS/MS, pharmacokinetics, chromatographic

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Authors: Jannika Dombrowski, Sabine Ambros, Ulrich Kulozik

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Due to the increasing popularity of healthy products, probiotics are still of rising importance in food manufacturing. With the aim to amplify the field of probiotic application to non-chilled products, the cultures have to be preserved by drying. Microwave drying has proved to be a suitable technique to achieve relatively high survival rates, resulting from drying at gentle temperatures, among others. However, diffusion limitation due to compaction of cell suspension during drying can prolong drying times as well as deteriorate product properties (grindability, rehydration performance). Therefore, we aimed to embed probiotics in an aerated matrix of whey proteins (surfactants) and di-/polysaccharides (foam stabilization, probiotic protection) during drying. As a result of the manifold increased inner surface of the cell suspension, drying performance was enhanced significantly as compared to non-foamed suspensions. This work comprises investigations on suitable foam matrices, being stable under vacuum (variation of protein concentration, type and concentration of di-/polysaccharide) as well as development of an applicable microwave drying process in terms of microwave power, chamber pressure and maximum product temperatures. Performed analyses included foam characteristics (overrun, drainage, firmness, bubble sizes), and properties of the dried cultures (survival, activity). In addition, efficiency of the drying process was evaluated.

Keywords: foam structure, microwave drying, polysaccharides, probiotics

Procedia PDF Downloads 262

Authors: Gina Zavaleta-Espejo, Segundo R. Jáuregui-Rosas, Fanny V. Samanamud-Moreno, José Saldaña Jiménez, Anibal Felix-Quintero, Víctor Montero-Del Aguila, Elsi Mejía-Uriarte

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Vicugnapacos "alpaca" fabrics are considered special for their finesse, and the garments in the textile market are very luxurious. It has many special characteristics such as antiallergic, soft, hygroscopic, among others. In this sense, the research aimed to evaluate the antimicrobial activity of alpaca fabrics functionalized with silver nanoparticles on the bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. For the functionalization of the fabrics, AgNO3 and different concentrations of trisodium citrate (TSC) 2, 6, and 10 mg. Tissue characterization was performed using Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The determination of the antimicrobial activity of the alpaca tissues was made by the Kirby-Bauer method with alpaca tissue discs functionalized with silver nanoparticles, an experimental design was made in completely randomized blocks with three treatments and a negative control with three repetitions. The results showed that inhibition halos were formed for both bacteria, therefore, the functionalized tissues have a high antimicrobial activity, whose mechanism of action is attributed to the free radicals (ROS) generated by the nanoparticles that cause oxidative damage to the bacteria. proteins and lipids of the bacterial cell wall.

Keywords: antimicrobial, animal fibers, fabrics, functionalization, trisodium citrate

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Authors: Agata Zakrzewska, Dominik Kopeć, Adrian Ochtyra

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The growing number of new cameras, sensors, and research methods allow for a broader application of thermal data in remote sensing vegetation studies. The aim of this research was to check whether it is possible to use thermal infrared data with a spectral range (3.6-4.9 μm) obtained during the day and the night to assess the health condition of selected species of deciduous trees in an urban environment. For this purpose, research was carried out in the city center of Warsaw (Poland) in 2020. During the airborne data acquisition, thermal data, laser scanning, and orthophoto map images were collected. Synchronously with airborne data, ground reference data were obtained for 617 studied species (Acer platanoides, Acer pseudoplatanus, Aesculus hippocastanum, Tilia cordata, and Tilia × euchlora) in different health condition states. The results were as follows: (i) healthy trees are cooler than trees in poor condition and dying both in the daytime and nighttime data; (ii) the difference in the canopy temperatures between healthy and dying trees was 1.06oC of mean value on the nighttime data and 3.28oC of mean value on the daytime data; (iii) condition classes significantly differentiate on both daytime and nighttime thermal data, but only on daytime data all condition classes differed statistically significantly from each other. In conclusion, the aerial thermal data can be considered as an alternative to hyperspectral data, a method of assessing the health condition of trees in an urban environment. Especially data obtained during the day, which can differentiate condition classes better than data obtained at night. The method based on thermal infrared and laser scanning data fusion could be a quick and efficient solution for identifying trees in poor health that should be visually checked in the field.

Keywords: middle wave infrared, thermal imagery, tree discoloration, urban trees

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Authors: Giuffrida Graziella, Pennisi Stefania, Coppa Federica, Iannello Giulia, Cartelli Simone, Lo Faro Riccardo, Ferruggia Greta, Brundo Maria Violetta

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Cladode chemical composition may vary according to soil factors, cultivation season, and plant age. The primary metabolites of cladodes are water, carbohydrates, and proteins. The carbohydrates in cladodes are divided into two types: structural and storage. Polysaccharides from Opuntia ficus‐indica (L.) Mill plants build molecular networks with the capacity to retain water; thus, they act as mucoprotective agents. Mucilage is the main polysaccharide of cladodes; it contains polymers of β‐d‐galacturonic acid bound in positions (1–4) and traces of R‐linked l‐rhamnose (1-2). Mucilage regulates both the cell water content during prolonged drought and the calcium flux in the plant cells. The in vitro analysis of keratinocytes in monolayer, through the scratch-wound-healing assay, provided promising results. After 48 hours of exposure, the wound scratch was almost completely closed in cells treated with cladode extract. After 72 hours, the treated cells reached complete confluence, while in the untreated cells (negative control) the confluence was reached after 96 hours. We also added a positive control group of cells treated with colchicine, which inhibited wound closure for a more comprehensive analysis.

Keywords: cladodes, metabolites, polysaccharide, scratch-wound-healing assay

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Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

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Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050, was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey

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Authors: Wahyu Purnama Putra, Artono Isharanto

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Thoracic trauma is trauma that hits the thoracic wall or intrathoracic organs, either due to blunt trauma or sharp trauma. Thoracic trauma often causes impaired ventilation-perfusion due to damage to the lung parenchyma. This results in impaired tissue oxygenation, which is one of the causes of acute respiratory distress syndrome (ARDS). These changes are caused by the release of pro-inflammatory mediators, plasmatic proteins, and proteases into the alveolar space associated with ongoing edema, as well as oxidative products that ultimately result in severe inhibition of the surfactant system. This study aims to predict the incidence of acute respiratory distress syndrome (ARDS) through human neutrophil elastase levels. This study examines the relationship between plasma elastase levels as a predictor of the incidence of ARDS in thoracic trauma patients in Malang. This study is an observational cohort study. Data analysis uses the Pearson correlation test and ROC curve (receiver operating characteristic curve). It can be concluded that there is a significant (p= 0.000, r= -0.988) relationship between elastase levels and BGA-3. If the value of elastase levels is limited to 23.79 ± 3.95, the patient will experience mild ARDS. While if the value of elastase levels is limited to 57.68 ± 18.55, in the future, the patient will experience moderate ARDS. Meanwhile, if the elastase level is between 107.85 ± 5.04, the patient will likely experience severe ARDS. Neutrophil elastase levels correlate with the degree of severity of ARDS incidence.

Keywords: ARDS, human neutrophil elastase, severity, thoracic trauma

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Authors: V. C. Santos-Ebinuma, J. F. B. Pereira, M. F. S. Teixeira, A. Pessoa Jr., P. Jauregi

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There is worldwide interest in process development for colorants production from natural sources. Microorganisms provide an alternative source of natural colorants which can be produced by cultivation technology and extracted from fermented broth. The aim of the present work was to study the recovery of red colorants from fermented broth of Penicillium purpurogenum DPUA 1275 using the technique of Colloidal Gas Aphrons (CGA); CGA are surfactant-stabilized microbubbles generated by intense stirring of a surfactant solution. CGA were generated by the cationic, hexadecyl trimethyl ammonium bromide (CTAB) surfactant. Firstly, experiments were carried out at different surfactant/fermented broth volumetric ratios (VCGA/VFB, VRATIO) varying between 3 and 18 at pH 6.9. Secondly, the experiments were carried out at VRATIO of 6 and 12 in different pH, namely, 6.9, 8.0, 9.0 and 10.0. The first results of recovery showed that an increase in the VRATIO from 3 to 6 and 12 promoted an increase as recovery as partition coefficient. However, at VRATIO of 18 the lowest partition coefficient was obtained. The best results were achieved at VRATIO of 6 and 12, namely recovery, Re, around 60% and partition coefficient, K, of 2.5 and 3.0 to 6 and 12 VRATIO, respectively. The second set of experiments showed that the pH 9.0 promoted the best results at VRATIO of 12 as follow: Re=70%, K=5.39, proteins and sugar selectivity (SePROT, 3.75 and SeSUGAR, 7.20, respectively). These results indicate that with CTAB the recovery is mainly driven by electrostatic interactions. In conclusion, the results above show that CGA employing a cationic surfactant is a promissory technique and it can be used as the first step of purification to recovery red colorants from fermented broth.

Keywords: liquid-liquid extraction, colloidal gas aphrons, recovery, natural colorants

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Authors: M. Youngsabanant-Areekijseree, C. Thepsithar, K. Sribuddhachart, J. Tananantayot

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Porcine oviductal fluid (pOF) from oviduct, an unused organ from the slaughterhouse, was effectively used for biotechnology studies. The fluid components consisted of micro- and macro-nutrients, amino acids, carbon source and proteins that played important roles in animal cell and embryo development. This was our knowledge on investigating pOF as growth promoting substance in culture medium of an orchid, Dendrobium mirbelianum. Two-leaf shoots were cultured in liquid Vacin and Went (VW) medium as a standard medium supplemented with 2 g/L peptone (Pe) or 100 g/ L boiled-potato water (Po) alone or in combinations, and added with 0, 1, 3 or 5 ml/L pOF. All explants were cultured in a stationary condition for 8 weeks. It was found that medium added with 100 g/L Po and 1 ml/L pOF provided the best results (1.02 g fresh weight, 4.2 shoots, 0.53 cm shoot height, 4.4 protocorms, 11.0 leaves and 5.7 roots with 100% survival) when compared to other medium, but not statistically significant difference from medium added with 100 g/L Po (0.86 g fresh weight, 4.3 shoots, 0.51 cm shoot height, 4.6 protocorms, 12.4 leaves and 6.6 roots with 100% survival). However, VW medium supplemented with 1 or 3 ml/L pOF alone showed the higher percentage of survival (100%) than VW medium (86.67%). It was shown the potential role of pOF as an organic supplement for promoting growth of plants. Acknowledgements—The project was funded by a grant from Silpakorn University Research & Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: Dendrobium mirbelianum, pig, oviductal fluid, in vitro growth

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Authors: Muhammad Jawad Saleem, Faisal Hafeez, Muhammad Arshad, Afifa Naeem, Ayesha Iftekhar

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Bacillus thuringiensis is a gram-positive spore-forming bacterium that belongs to the Bacillus cereus group of Bacilli and it produces ICP (insecticidal crystal protein) Cry toxins or Cysts toxins. Spores are produced as parasporal crystalline inclusions bodies (also known as endotoxins) at the onset of sporulation during the stationary growth phase. During vegetative growth that does not form crystals and is called vegetative insecticidal proteins (VIP) and secreted an insecticidal protein (SIP). Bacillus thuringiensis (Bt) is important for pest management either in the form of insecticides or through incorporated in the gene of the crop. Bioassays were conducted on the F2 generation of 1st instar larvae of H. armigera by the diet incorporation method to determine the susceptibility to Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The median lethal concentration (LC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.11 to 1.06 µg/ml and moult inhibitory concentration (MIC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.05 to 0.25 µg/ml. Cry1Ac was found most toxic to 1st instar larvae of H. armigera as compared to other Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The experimental results are important to policy-makers and technology providers to develop strategies for the exploitation of transgenic Bt cotton varieties as a component of integrated pest management.

Keywords: Bt toxin, Cry1Ac, Cry2Ab, Cry2A, susceptibility, Helicoverpa armigera

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Authors: Adewole Tomiwa Adetunji, Francis Bayo Lewu, Richard Mundembe

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Plants require nitrogen (N) to support desired production levels. There is a need for better understanding of N transport mechanism in order to improve N assimilation by plant root. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which N is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was amplified, sequenced and characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design 976 base pairs AMT1-specific primers which include forward primer 5’- GCCATCGCCGCCGCCGG-3’ and reverse primer 5’-GGGTCAGATCCATACCCGC-3’. These primers were used to amplify the Solanum tuberosum AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family due to the clade and high similarity it shared with other plant AMT1 genes. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1, and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th-10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

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Authors: Carmela Perta

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Among the studies flourished in the field of historical sociolinguistics, mainly in the strand devoted to English history, during its Medieval and early modern phases, multilingual texts had been analysed using theories and models coming from contact linguistics, thus applying synchronic models and approaches to the past. This is true also in the case of contact phenomena which would transcend the writing level involving the language systems implicated in contact processes to the point of perceiving a new variety. This is the case for medieval administrative-commercial texts in which, according to some Scholars, the degree of fusion of Anglo-Norman, Latin and middle English is so high a mixed code emerges, and there are recurrent patterns of mixed forms. Interesting is a collection of multilingual business writings by John Balmayn, an Englishman overseeing a large shipment in Tuscany, namely the Cantelowe accounts. These documents display various analogies with multilingual texts written in England in the same period; in fact, the writer seems to make use of the above-mentioned patterns, with Middle English, Latin, Anglo-Norman, and the newly added Italian. Applying an atomistic yet dynamic approach to the study of contact phenomena, we will investigate these documents, trying to explore the nature of the switching forms they contain from an intra-writer variation perspective. After analysing the accounts and the type of multilingualism in them, we will take stock of the assumed mixed code nature, comparing the characteristics found in this genre with modern assumptions. The aim is to evaluate the possibility to consider the switching forms as core elements of a mixed code, used as professional variety among merchant communities, or whether such texts should be analysed from a switching perspective.

Keywords: historical sociolinguistics, historical code switching, letters, medieval england

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Authors: Deepak Mohan, Sushma Sharma, Mohammad Asif

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Diclofenac sodium being one of the most common non-steroidal anti-inflammatory drugs is normally used as painkiller and to reduce inflammation. The drug is known to alter the enzymatic activities of acid and alkaline phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminases. The drug also results in change in the concentration of proteins and lipids in the body. The present study is an attempt to study different biochemical changes electrophoretically due to administration of different doses of diclofenac (4mg/kg/body weight and 14mg/kg/body weight) on liver and testes of mice from 7-28 days of investigation. Homogenization of the tissue was done, supernatant separated was loaded in the gel and native polyacrylamide gel electrophoresis was conducted. Diclofenac administration resulted in alterations of all these biochemical parameters which were observed in native polyacrylamide gel electrophoretic studies. The severe degenerative changes as observed during later stages of the experiment showed correlation with increase or decrease in the activities of all the enzymes studied in the present investigation. Image analysis of gel in liver showed a decline of 7.4 and 5.3 % in low and high dose group after 7 days whereas a decline of 9.6 and 7.5% was registered after 28 days of investigation. Similar analysis for testis also showed an appreciable decline in the activity of alkaline phosphatase after 28 days. Gel analysis of serum was also performed to find a correlation in the enzymatic activities between the tissue and blood.

Keywords: diclofenac, inflammation, polyacrylamide, phosphatase

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Authors: Cahit Demetgul, Sahin Bayraktar, Neslihan Beyazit

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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

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Authors: Matthew Ferguson, Tatyana Konkova, Ioannis Violatos

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Microstructure analysis of additively manufactured (AM) materials is an important step in understanding the interrelationship between mechanical properties and materials performance. Literature on the effect of laser-based AM process parameters on the microstructure in the substrate-deposit interface is limited. The interface region, the adjoining area of substrate and deposit, is characterized by the presence of the fusion zone (FZ) and heat-affected zone (HAZ), experiencing rapid thermal gyrations resulting in thermal-induced transformations. Inconel 718 was utilized as work material for both the substrate and deposit. Three blocks of Inconel 718 material were deposited by Direct Energy Deposition (DED) using three different laser powers, 550W, 750W and 950W, respectively. A coupled thermo-mechanical transient approach was utilized to correlate temperature history to the evolution of microstructure. The thermal history of the deposition process was monitored with the thermocouples installed inside the substrate material. The interface region of the blocks was analyzed with Optical Microscopy (OM) and Scanning Electron Microscopy (SEM), including the electron back-scattered diffraction (EBSD) technique. Laser power was found to influence the dissolution of intermetallic precipitated phases in the substrate and grain growth in the interface region. Microstructure and thermal history data were utilized to draw conclusive comparisons between the investigated process parameters.

Keywords: additive manufacturing, direct energy deposition, electron back-scattered diffraction, finite element analysis, inconel 718, microstructure, optical microscopy, scanning electron microscopy, substrate-deposit interface region

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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

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Authors: Shubham Dwivedi, Raghavender Medishetti, Rita Rani, Aarti Sevilimedu, Pushkar Kulkarni, Yogeeswari Perumal

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Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder of early onset, characterized by impaired sociability, cognitive function and stereotypies. There is a significant urge to develop and establish new animal models with ASD-like characteristics for better understanding of underlying mechanisms. The aim of the present study was to develop a cost and time effective zebrafish model with quantifiable parameters to facilitate mechanistic studies as well as high-throughput screening of new molecules for autism. Zebrafish embryos were treated with valproic acid and a battery of behavioral tests (anxiety, inattentive behavior, irritability and social impairment) was performed on larvae at 7th day post fertilization, followed by study of molecular markers of autism. This model shows a significant behavioural impairment in valproic acid treated larvae in comparison to control which was again supported by alteration in few marker genes and proteins of autism. The model also shows a rescue of behavioural despair with positive control drugs. The model shows robust parameters to study behavior, molecular mechanism and drug screening approach in a single frame. Thus we postulate that our 7 days zebrafish larval model for autism can help in high throughput screening of new molecules on autism.

Keywords: autism, zebrafish, valproic acid, neurodevelopment, behavioral assay

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Authors: Mohd Adil, Rosina Khan, Danishuddin, Asad U. Khan

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The aim of this study was to evaluate the influence of the crude and active solvent fraction of Trachyspermum ammi on S. mutans cariogenicity, effect on expression of genes involved in biofilm formation and caries development in rats. GC–MS was carried out to identify the major components present in the crude and the active fraction of T. ammi. The crude extract and the solvent fraction exhibiting least MIC were selected for further experiments. Scanning electron microscopy was carried out to observe the effect of the extracts on S. mutans biofilm. Comparative gene expression analysis was carried out for nine selected genes. 2-Isopropyl-5-methyl-phenol was found as major compound in crude and the active fraction. Binding site of this compound within the proteins involved in biofilm formation was mapped with the help of docking studies. Real-time RT-PCR analyses revealed significant suppression of the genes involved in biofilm formation. All the test groups showed reduction in caries (smooth surface as well as sulcal surface caries) in rats. Moreover, it also provides new insight to understand the mechanism influencing biofilm formation in S. mutans. Furthermore, the data suggest the putative cariostatic properties of T. Ammi and hence can be used as an alternative medicine to prevent caries infection.

Keywords: bio-film, Streptococcus mutans, dental caries, bio-informatic

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Authors: Yaya Gao, Jifeng Liu, Yafeng Liu

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Objectives: This study aimed to improve clinicians' understanding and diagnosis of the Mammary analogue secretory carcinoma of the salivary gland（MASC）. Methods: The clinical features of a MASC patient who was admitted to WestChina Hospital of Sichuan University in July 2020 were reviewed and analyzed. A 49-year-old woman with left upper neck pain for three months was admitted to the hospital. She underwent adenoma resection of the left submandibular gland 14 years ago and mucoepidermoid carcinoma resection surgery five years ago. Three months before admission, the patient developed pain in the left mandibular angle after "fatigue" and gradually developed radiation pain in the left ear, which could be relieved after rest. A mass of 1cm could be touched at the mandibular, with tenderness, poor mobility, and hard texture. No swelling, heat, pain, rupture, or pus was found on the surrounding skin. Color doppler ultrasonography of the salivary gland indicated a weak echo mass of 23*14*17mm in the left parotid gland. Results: Surgical excision was completed. Immunohistochemistry of the tumor samples after operation showed that P63（a few，+）, CK7（+）, S100（+）, DOG1（-）, Ki67（MIB-1）（+，5%），pan-TRK（+）, PAS(+) . ETV-6 gene translocation was detected in FISH in postoperative pathology, which indicated MASC. After this diagnosis, the patient sent the postoperative specimen of the second submandibular tumor to our hospital for consultation. The morphology of the two was similar. FISH detected ETV-6 gene translocation, so the second pathological diagnosis was revised to MASC. Conclusion: MASC of the salivary gland is a rare salivary gland tumor whose diagnosis depends on the result of the ETV6-NTRK3 fusion gene.

Keywords: mammary analogue secretory carcinoma, ETV6-NTRK3, salivary gland, misdiagnosed

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Authors: Muhammad H. Alu’datt, Inteaz Alli

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This study was conducted to prepare and evaluate the biological properties of protein co-precipitates from flaxseed and soybean. Protein was prepared by NaOH extraction through the mixing of soybean flour (Sf) and flaxseed flour (Ff) or mixtures of soybean extract (Se) and flaxseed extract (Fe). The protein co-precipitates were precipitated by isoelectric (IEP) and isoelectric-heating (IEPH) co-precipitation techniques. Effects of extraction and co-precipitation techniques on co-precipitate yield were investigated. Native-PAGE, SDS-PAGE were used to study the molecular characterization. Content and antioxidant activity of extracted free and bound phenolic compounds were evaluated for protein co-precipitates. Removal of free and bound phenolic compounds from protein co-precipitates showed little effects on the electrophoretic behavior of the proteins or the protein subunits of protein co-precipitates. Results showed that he highest protein contents and yield were obtained in for Sf-Ff/IEP co-precipitate with values of 53.28 and 25.58% respectively as compared to protein isolates and other co-precipitates. Results revealed that the Sf-Ff/IEP showed a higher content of bound phenolic compounds (53.49% from total phenolic content) as compared to free phenolic compounds (46.51% from total phenolic content). Antioxidant activities of extracted bound phenolic compounds with and without heat treatment from Sf-Ff/IEHP were higher as compared to free phenolic compounds extracted from other protein co-precipitates (29.68 and 22.84%, respectively).

Keywords: antioxidant, phenol, protein co-precipitate, yield

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Authors: Bheeshma B., Mathivanan N., Manoj Deepak M., Prabhu Thangaraju, K. Venkatachalam

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This study involves the effect of autologous blood injection for patients who had degeneration of the origin of extensor carpi radialis brevis which was confirmed radio logically and by ultrasound examination and failed cortisone injections to the lateral epicondylitis. In this prospective longitudinal series involves pre-injection assessment of grip strength, pain, and function, using the patient-rated tennis elbow evaluation. In this study, blood from the contralateral limb is taken and injected into the affected limb with the help of ultrasound guidance and then the patient wore a customized wrist support for five days, after which they were commenced with stretching, strengthening, and massage programme with an occupational therapist. In these patients assessment was done after six months and then finally at 12 months after injection, using the patient-rated tennis elbow evaluation. 50 patients completed the study, showing significant improvement in pain; the worst pain decreased by two to five points out of a 10-point visual analogue for pain. Self-perceived function improved by 11–25 points out of 100. Women showed significant increase in grip, but men did not. Our study thus concludes that autologous blood injection show significant improvement in pain and function in patients with chronic lateral epicondylitis, who did not have relief with cortisone injection.

Keywords: lateral epicondylitis, autologous blood injection, conservative treatment, plasma-rich proteins (PRPs)

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Authors: Joan O. Ogundele, Aladesanmi A. Oshodi, Adekunle I. Amoo

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Abstract In-vitro multi-enzyme protein digestibility (IVMPD) and some anti-nutritional factors (ANF) of five melon (egusi) seed flours (MSF) and their protein isolates (PI) were carried out. Their PI have potentials comparable to that of soya beans. It is important to know the IVMPD and ANF of these protein sources as to ensure their safety when adapted for use as alternate protein sources to substitute for cow milk, which is relatively expensive in Nigeria. Standard methods were used to produce PI of Citrullus colocynthis, Citrullus vulgaris, African Wine Kettle gourd (Lageneria siceraria I), Basket Ball gourd (Lagenaria siceraria II) and Bushel Giant Gourd (Lageneria siceraria III) seeds and to determine the ANF and IVMPD of the MSF and PI unheated and at 37oC. Multi-enzymes used were trypsin, chymotrypsin and peptidase. IVMPD of MSF ranged from (70.67±0.70) % (C. vulgaris) to (72.07± 1.79) % (L.siceraria I) while for their PI ranged from 74.33% (C.vulgaris) to 77.55% (L.siceraria III). IVMPD of the PI were higher than those of MSF. Heating increased IVMPD of MSF with average value of 79.40% and those of PI with average of 84.14%. ANF average in MSF are tannin (0.11mg/g), phytate (0.23%). Differences in IVMPD of MSF and their PI at different temperatures may arise from processing conditions that alter the release of amino acids from proteins by enzymatic processes. ANF in MSF were relatively low, but were found to be lower in the PI, therefor making the PI safer for human consumption as an alternate source of protein.

Keywords: Anti-nutrients, Enzymatic protein digestibility, Melon (egusi)., Protein Isolates.

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