Search results for: enzyme hydrolysis

Authors: Joanna Cabaj, Sylwia Baluta, Karol Malecha

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Catecholamines are vital neurotransmitters that mediate a variety of central nervous system functions, such as motor control, cognition, emotion, memory processing, and endocrine modulation. Dysfunctions in catecholamine neurotransmission are induced in some neurologic and neuropsychiatric diseases. Changeable neurotransmitters level in biological fluids can be a marker of several neurological disorders. Because of its significance in analytical techniques and diagnostics, sensitive and selective detection of neurotransmitters is increasingly attracting a lot of attention in different areas of bio-analysis or biomedical research. Recently, optical techniques for the detection of catecholamines have attracted interests due to their reasonable cost, convenient control, as well as maneuverability in biological environments. Nevertheless, with the observed need for a sensitive and selective catecholamines sensor, the development of a convenient method for this neurotransmitter is still at its basic level. The manipulation of nanostructured materials in conjunction with biological molecules has led to the development of a new class of hybrid-modified enzymatic sensors in which both enhancement of charge transport and biological activity preservation may be obtained. Immobilization of biomaterials on electrode surfaces is the crucial step in fabricating electrochemical as well as optical biosensors and bioelectronic devices. Continuing systematic investigation in manufacturing of enzyme–conducting sensitive systems, here is presented a convenient fluorescence as well as electrochemical sensing strategy for catecholamines detection.

Keywords: biosensors, catecholamines, fluorescence, enzymes

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Authors: Heriansyah Putra, Erizal Erizal, Sutoyo Sutoyo, Hideaki Yasuhara

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This paper discusses the possibility of organic waste material, i.e., soybean, as the bio-catalyst agent on the calcite precipitation method. Several combinations of soybean powder and jack bean extract are used as the bio-catalyst and mixed with the reagent composed of calcium chloride and urea. Its productivity in promoting calcite crystal is evaluated through a transparent test-tube experiment. The morphological and mineralogical aspects of precipitated calcite are also investigated using scanning electromagnetic (SEM) and X-ray diffraction (XRD), respectively. The applicability of this material to improve the engineering properties of soil are examined using the direct shear and unconfined compressive test. The result of this study shows that the utilization of soybean powder brings about a significant effect on soil strength. In addition, the use of soybean powder as a substitution material of urease enzyme also increases the efficacy of calcite crystal as the binder materials. The low calcite content promotes the high strength of the soil. The strength of 300 kPa is obtained in the presence of 2% of calcite content within the soil. The result of this study elucidated that substitution of soybean to jack bean extract is the potential and valuable alternative to improve the applicability of calcite precipitation method as soil improvement technique.

Keywords: calcite precipitation, jack bean, soil improvement, soybean

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Authors: Ibrahim Khalil Ibrahim, Enas Mohamed Shahine, Abeer Shawky El Hadedy, Emmanuel Kamal Aziz Saba, Ghada Salah Attia Hussein

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Osteoarthritis (OA) is a multifactorial disease characterized by a progressive degradation of articular cartilage and is the leading cause of disability in elderly persons. IL-18 contributes to the destruction of cartilage and bone in the disease process of arthritis. The aim of the study was to investigate the role of IL-18 in primary knee OA patients. Serum level of IL-18 was assessed by enzyme-linked immunosorbent assay in 30 primary knee OA patients and compared to 20 age and gender-matched healthy volunteers as a control group. Radiographic severity of OA was assessed by Kellgren and Lawrence (KL) global scale. Pain, stiffness and functional assessment were done using Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). OA patients had significantly higher serum IL-18 level than in control group (420.93 ± 345.4 versus 151.03 ± 144.16 pg/ml, P=0.001). Serum level of IL-18 was positively correlated with KL global scale (P=0.001). There were no statistically significant correlations between serum level of IL-18 and pain, stiffness, function subscales and total WOMAC index scores among the studied patients. In conclusions, IL-18 has a role in the pathogenesis of OA and it is positively correlated with the radiographic damage of OA.

Keywords: Interlukin-18, knee osteoarthritis, primary osteoarthritis, WOMAC scale

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Authors: Ge Cui, Mei Fang Chien, Chihiro Inoue

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In this research, enrichment culture using an inorganic liquid medium collected soil contaminated with 1,2-dichlorobenzene (1,2-DCB) in Sendai, Japan, was added 1,2-DCB as the sole carbon source to create a stable consortium. The purpose of this research is to analysis dominant microorganisms in the stable consortium and enzyme system which play a role in the degradation of DCBs. The consortium is now at 30 generation and is still being cultured. By the result of PCR-DGGE and clone library, two bacteria are dominant. The bacteria named sk1 was isolated. 40mg/l of 1,2-DCB and 40mg/l of 1,4-DCB were completely degraded after 32 hours and 50 hours, respectively, but no degradation occurred in the case of 1,3-DCB. By PCR, tecA1 (α-subunit of DCB dioxygenase) gene which plays a role degrading DCB to DCB dihydrodiol, and tecB (dehydrogenase) gene which plays a role degrading DCB dihydrodiol to dichlorocatechol were amplified from strain sk1. Bacteria named sk100 was also isolated. 40mg/l of 1,2-DCB was completely degraded after 32 hours, but no degradation occurred in case of 1,3-DCB and 1,4-DCB. By the result of the catalytic core region of dioxygenase amplified by PCR, gene played a role degrading DCB was analyzed. The results of this study concluded that the isolated strains which have not been reported are able to degrade 1,2-DCB stably, and the characterization of degradation and the genomic analysis which is now in progress is helpful to have an overall view of this microbial degradation.

Keywords: DCB, 1, 2-DCB degrading strains, DCB dioxygenase, enrichment culture

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Authors: Virginia Martin Torrejon, Binjie Wu

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Gelatine is a protein biopolymer obtained from the partial hydrolysis of animal tissues which contain collagen, the primary structural component in connective tissue. Gelatine hydrogels have attracted considerable research in recent years as an alternative to synthetic materials due to their outstanding gelling properties, biocompatibility and compostability. Surfactants, such as sodium dodecyl sulfate (SDS), are often used in hydrogels solutions as surface modifiers or solubility enhancers, and their incorporation can influence the hydrogel’s viscoelastic properties and, in turn, its processing and applications. Literature usually focuses on studying the impact of formulation parameters (e.g., gelatine content, gelatine strength, additives incorporation) on gelatine hydrogels properties, but processing parameters, such as curing temperature, are commonly overlooked. For example, some authors have reported a decrease in gel strength at lower curing temperatures, but there is a lack of research on systematic viscoelastic characterisation of high strength gelatine and gelatine-SDS systems at a wide range of curing temperatures. This knowledge is essential to meet and adjust the technological requirements for different applications (e.g., viscosity, setting time, gel strength or melting/gelling temperature). This work investigated the effect of curing temperature (10, 15, 20, 23 and 25 and 30°C) on the elastic modulus (G’) and melting temperature of high strength gelatine-SDS hydrogels, at 10 wt% and 20 wt% gelatine contents, by small-amplitude oscillatory shear rheology coupled with Fourier Transform Infrared Spectroscopy. It also correlates the gel strength obtained by rheological measurements with the gel strength measured by texture analysis. Gelatine and gelatine-SDS hydrogels’ rheological behaviour strongly depended on the curing temperature, and its gel strength and melting temperature can be slightly modified to adjust it to given processing and applications needs. Lower curing temperatures led to gelatine and gelatine-SDS hydrogels with considerably higher storage modulus. However, their melting temperature was lower than those gels cured at higher temperatures and lower gel strength. This effect was more considerable at longer timescales. This behaviour is attributed to the development of thermal-resistant structures in the lower strength gels cured at higher temperatures.

Keywords: gelatine gelation kinetics, gelatine-SDS interactions, gelatine-surfactant hydrogels, melting and gelling temperature of gelatine gels, rheology of gelatine hydrogels

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Authors: Tripti Mishra, Shipra Shukla, Sanjeev Meena, , Ruchi Singh, Mahesh Pal, D. K. Upreti, Dipak Datta

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Roccella montagnei belongs to lichen family Roccelleceae growing luxuriantly along the coastal regions of India. As Roccella has been shown to be bioactive, we prepared methanolic extract and assessed its anticancer potential. The methanolic extract showed significant in vitro cytotoxic activity against four human cancer cell lines such as Colon (DLD-1, SW-620), Breast (MCF-7), Head and Neck (FaDu). This prompted us to isolate bioactive compounds through column chromatography. Two compounds Roccellic acid and Everninic acid have been isolated, out of which Everninic acid is reported for the first time. Both the compounds have been tested for in vitro cytotoxic activity in which Roccellic acid showed strong anticancer activity as compared to the Everninic acid. CDK-10 (Cyclin-dependent kinase) contributes to proliferation of cancer cells, and aberrant activity of these kinases has been reported in a wide variety of human cancers. These kinases, therefore, constitute biomarkers of proliferation and attractive pharmacological targets for the development of anticancer therapeutics. Therefore both the isolated compounds were tested for in silico molecular docking study against CDK-10 isomer enzyme to support the cytotoxic activity.

Keywords: cytotoxic activity, everninic acid, roccellic acid, R. montagnei

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Authors: Reza Ferdousi, Reza Safdari, Yadollah Omidi

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Drug-drug interactions (DDIs) are main cause of the adverse drug reactions and nature of the functional and molecular complexity of drugs behavior in human body make them hard to prevent and treat. With the aid of new technologies derived from mathematical and computational science the DDIs problems can be addressed with minimum cost and efforts. Market basket analysis is known as powerful method to identify co-occurrence of thing to discover patterns and frequency of the elements. In this research, we used market basket analysis to identify important bio-elements in DDIs occurrence. For this, we collected all known DDIs from DrugBank. The obtained data were analyzed by market basket analysis method. We investigated all drug-enzyme, drug-carrier, drug-transporter and drug-target associations. To determine the importance of the extracted bio-elements, extracted rules were evaluated in terms of confidence and support. Market basket analysis of the over 45,000 known DDIs reveals more than 300 important rules that can be used to identify DDIs, CYP 450 family were the most frequent shared bio-elements. We applied extracted rules over 2,000,000 unknown drug pairs that lead to discovery of more than 200,000 potential DDIs. Analysis of the underlying reason behind the DDI phenomena can help to predict and prevent DDI occurrence. Ranking of the extracted rules based on strangeness of them can be a supportive tool to predict the outcome of an unknown DDI.

Keywords: drug-drug interaction, market basket analysis, rule discovery, important bio-elements

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Authors: Francesco Angelucci, Katerina Veverova, Alžbeta Katonová, Lydia Piendel, Martin Vyhnalek, Jakub Hort

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Alzheimer's disease (AD) is a central nervous system (CNS) disease characterized by loss of memory, cognitive functions and neurodegeneration. Plasmin is an enzyme degrading many plasma proteins. In the CNS, plasmin may reduce the accumulation of A, and have other actions relevant to AD pathophysiology. Brain plasmin synthesis is regulated by two enzymes: one activating, the tissue plasminogen activator (tPA), and the other inhibiting, the plasminogen activator inhibitor-1 (PAI-1). We investigated whether tPA and PAI-1 serum levels in AD and amnestic mild cognitive impairment (aMCI) patients are altered compared to cognitively healthy controls. Moreover, we examined the PAI-1/tPA ratio in these patient groups. 40 AD, 40 aMCI and 10 healthy controls were recruited. Venous blood was collected and PAI-1 and tPA serum concentrations were quantified by sandwich ELISAs. The results showed that PAI-1 levels increased in AD and aMCI patients. This increase negatively correlated with cognitive deficit measured by MMSE. Similarly, the ratio between tPA and PAI-1 gradually increases in aMCI and AD patients. This study demonstrates that AD and aMCI patients have altered PAI-1 serum levels and PAI-1/tPA ratio. Since these enzymes are CNS regulators of plasmin, PAI-1 serum levels could be a marker reflecting a cognitive decline in AD.

Keywords: Alzheimer disease, amnestic mild cognitive impairment, plasmin, tissue-type plasminogen activator

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Authors: Kai-Cheng Hsu, Tzu-Ying Sung, Jinn-Moon Yang

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Colorectal cancer (CRC) is one of the main causes of cancer death in the world. Although several drugs have been developed to treat colorectal cancer, such as Regorafenib and 5-FU, their efficacy is often limited by the development of drug resistance. Therefore, development of new drugs with new scaffolds is necessary to treat CRC. Here, we used site-moiety maps to identify inhibitors against PIM1, LIMK1, SRC, and mTOR, which are often overexpressed in CRC. A site-moiety map represents physicochemical properties and moiety preferences of a binding site through anchors. An anchor contains three elements: (1) conserved interacting residues of a binding pocket; (2) moiety preference of the binding pocket; and (3) the type (e.g., hydrogen-bonding or van der Waals interactions) of interaction between the moieties and the binding pocket. Then, we performed a structure-based virtual screening of ~260,000 compounds and selected compound candidates with high site-moiety map scores for bioassays. Among these candidates, compound 1 and compound 2 inhibited the growth of CRC cells with IC50 values of <10 μM. The experimental result of enzyme-based assays indicated that compound 1 is a dual inhibitor against PIM1 (IC50 6 μM) and LIMK1(IC50 11 μM). Compound 2 was predicted as a SRC inhibitor and will be further validated. The compounds inhibited different protein targets compared to the current drugs. We believe that the compounds provide a starting point to design new drugs for CRC treatment.

Keywords: colorectal cancer, drug discovery, site-moiety map, virtual screening, PIM1, LIMK1

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Authors: Anusha Bhardwaj, Shekhar Shinde

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Nitric oxide (NO•) has been documented in research papers as one of the most versatile player in the therapeutics. It is identified as a biological multifunctional messenger molecule which is synthesized by the action of nitric oxide synthase (NOS) enzyme from L-arginine. The protective and the toxic effect in conjunction form the complete picture of the biological function of nitric oxide in humans. The dual nature is because of various factors such as concentration of NO, the isoform of NOS involved, type of cells in which it is synthesized, reaction partners like proteins, reactive oxygen intermediates, prosthetic groups, thiols etc., availability of the substrate L-arginine, intracellular environment in which NO is produced and generation of guanosine 3, 5’- cyclic monophosphate (cGMP). Activation of NOS through infection or trauma leads to one or more systemic effects including enhanced immune activity against invading pathogens, vaso/bronchodilatation in the cardiovascular and respiratory systems and altered neurotransmission which can be protective or toxic. Hence, NO affects the balance between healthy signaling and neurodegeneration in the brain. In lungs, it has beneficial effects on the function of airways as a bronchodilator and acts as the neurotransmitter of bronchodilator nerves. Whereas, on the other hand, NO may have deleterious effects by amplifying the asthmatic inflammatory response and also act as a vasodilator in the airways by increasing plasma exudation. But NOS Inhibitors and NO donors hamper the signalling pathway and hence a therapeutic application of NO is compromised.

Keywords: nitric oxide, multifunctional, dual nature, therapeutic applications

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Authors: Anjali Verma, Mahesh Pal, Veena Pande, Dalip Kumar Upreti

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The present study is carried out to explore the anti-hyperglycemic activity of Leucas cephalotes plant parts. A fruit, leaves, stems, and roots part of the Leucas cephalotes has been extracted in ethanol and have been evaluated for anti-hyperglycemic activity. The present study indicated that, ethanolic extract of fruit and leaves have shown significant α- amylase inhibitory activity with IC50 value of 92.86 ± 0.89 μg/mL and 98.09 ± 0.69 μg/mL respectively. Two known compounds β-sitosterol and lupeol were isolated from ethanolic extract of L. cephalotes leaves and were subjected to anti-hyperglycemic activity. Lupeol shows the best activity with IC50 55.73 ± 0.47 μg/mL and the results were verified by docking study of these compounds with mammalian α-amylase was carried out on its active site. It was concluded from the study that β-sitosterol and lupeol form one H-bond interactions with the active site residues either Asp212 or Thr21. The estimated free energy binding of β-sitosterol was found to be -9.47 kcal mol-1 with an estimated inhibition constant (Ki) of 558.94 nmol whereas the estimated free energy binding of lupeol was -11.73 kcal mol-1 with an estimated inhibition constant (Ki) of 476.71pmmol. The present study clearly showed that lupeol is more potent in comparison to β-sitosterol. The study indicates that L. cephalotes have significant potential to inhibit α-amylase enzyme.

Keywords: alpha-amylase, beta-sitosterol, hyperglycemia, lupeol

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Authors: S. S. Patil, R. M. Mhetre, S. V. Patil

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Lisinopril is an angiotensin converting enzyme inhibitor used in the treatment of hypertension and heart failure in prophylactic treatment after myocardial infarction and in diabetic nephropathy. However, it is very poorly absorbed from gastro-intestinal tract. Intranasal administration is an ideal alternative to the parenteral route for systemic drug delivery. Formulating multiparticulate system with mucoadhesive polymers provide a significant increase in the nasal residence time. The aim of the present approach was to overcome the drawbacks of the conventional dosage forms of lisinopril by formulating intranasal microspheres with Carbopol 974P NF and HPMC K4 M along with film forming polymer ethyl cellulose.The microspheres were prepared by emulsion solvent evaporation method. The prepared microspheres were characterized for encapsulation efficiency, drug loading, particle size, and surface morphology, degree of swelling, ex vivo mucoadhesion, drug release, ex vivo diffusion studies. All formulations has shown entrapment efficiency between 80 to more than 95%, mucoadhesion was more than 80 % and drug release up to 90 %. Ex vivo studies revealed tht the improved bioavailability of drug compared to oral drug administration. Both in vitro and in vivo studies conclude that combination of Carbopol and HPMC based microspheres shown better results than single carbopol based microspheres for the delivery of lisinopril.

Keywords: microspheres, lisinopril, nasal delivery, solvent evaporation method

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Authors: Syed M. Shahid, Rozeena Shaikh, Syeda N. Nawab, Abid Azhar

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Diabetes mellitus (DM) is a prevalent non-communicable disease worldwide. DM may lead to many vascular complications like hypertension, nephropathy, retinopathy, neuropathy and foot infections. Pathogenesis of diabetic nephropathy (DN) is implicated by the polymorphisms in genes encoding the specific components of renin angiotensin aldosterone system (RAAS) which include angiotensinogen (AGT), angiotensin-II receptor and angiotensin converting enzyme (ACE) genes. This study was designed to explore the possible association of AG (M268T) polymorphism in the patients of diabetes and nephropathy in Pakistan. Study subjects included 100 controls, 260 diabetic patients without renal insufficiency and 190 diabetic nephropathy patients with persistent albuminuria. Fasting blood samples were collected from all the subjects after getting institutional ethical approval and informed consent. The biochemical estimations, PCR amplification and direct sequencing for the specific region of AGT gene was carried out. A significantly high frequency of TT genotype and T allele of AGT (M268T) was observed in the patients of diabetes with nephropathy as compared to controls and diabetic patients without any known renal impairment. The TT genotype and T allele of AGT (M268T) polymorphism may be considered as a genetic risk factor for the development and progression of nephropathy in diabetes. Further cross sectional population studies would be of help to establish and confirm the observed possible association of AGT gene variations with development of nephropathy in diabetes.

Keywords: RAAS, AGT (M268T), diabetes, nephropathy

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Authors: Manuela Amorim, Cláudia S. Marques, Maria Conceição Calhau, Hélder J. Pinheiro, Maria Manuela Pintado

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Recently it has been recognized peptides from different food sources with biological activities. However, no relevant study has proven the potential of brewer yeast peptides in the modulation of gut microbiota. The importance of human intestinal microbiota in maintaining host health is well known. Probiotics, prebiotics and the combination of these two components, can contribute to support an adequate balance of the bacterial population in the human large intestine. The survival of many bacterial species inhabiting the large bowel depends essentially on the substrates made available to them, most of which come directly from the diet. Some of these substrates can be selectively considered as prebiotics, which are food ingredients that can stimulate beneficial bacteria such as Lactobacilli or Bifidobacteria growth in the colon. Moreover, conventional food can be used as vehicle to intake bioactive compounds that provide those health benefits and increase people well-being. In this way, the main objective of this work was to study the potential prebiotic activity of brewer yeast peptide extract (BYP) obtained via hydrolysis of yeast proteins by cardosins present in Cynara cardunculus extract for possible use as a functional ingredient. To evaluate the effect of BYP on the modulation of gut microbiota in diet-induced obesity model, Wistar rats were fed either with a standard or a high-fat diet. Quantified via 16S ribosomal RNA (rRNA) expression by quantitative PCR (qPCR), genera of beneficial bacteria (Lactobacillus spp. and Bifidobacterium spp.) and three main phyla (Firmicutes, Bacteroidetes and Actinobacteria) were assessed. Results showed relative abundance of Lactobacillus spp., Bifidobacterium spp. and Bacteroidetes was significantly increased (P < 0.05) by BYP. Consequently, the potential health-promoting effects of WPE through modulation of gut microbiota were demonstrated in vivo. Altogether, these findings highlight the possible intervention of BYP as gut microbiota enhancer, promoting healthy life style, and the incorporation in new food products, leads them bringing associated benefits endorsing a new trend in the improvement of new value-added food products.

Keywords: functional ingredients, gut microbiota, prebiotics, brewer yeast peptide extract

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Authors: Rajni Kant Panik, Deependra Singh, Manju Singh

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Despite significant advances in the treatment of severe burn injury, infection and sepsis persist as frequent causes of morbidity and mortality for burn victims due to extensive compromise of the skin and contiguous tissue that serve as a protective barrier against microbial invasion. In the setting of a burn wound infection, Staphylococcus aureus is the most commonly isolated pathogens from bloodstream infections in burn care hospitals. We aimed to develop a biocompatible system of Poly vinyl alcohol (PVA)-sodium alginate hydrogel carrying multiple drugs- catalase and mupirocin in controlled manner for effective and complete burn wound healing. PLGA nanoparticles of Catalase and mupirocin were prepared by homogenization method and optimized system was incorporated in PVA-sodium alginate slurry. PVA-sodium alginate hydrogels were prepared by freeze thaw method. The prepared dispersion was casted into films to prepare multiphase hydrogel system and characterized by in vitro and in vivo studies. The study clearly showed the beneficial effect of antioxidant enzyme and antibiotic in the treatment of infected burn wound, as evidenced by the reduced incidence of wound infection and the shortening of healing time.

Keywords: burn wound, catalase, mupirocin, wound healing

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Authors: Z. Mmango, U. Nwodo, L. V. Mabinya, A. I. Okoh

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Cellulose and hemicellulose account for a large portion of the world‘s plant biomass. In nature, these polysaccharides are intertwined forming complex materials that requires multiple and expensive treatment processes to free up the raw materials trapped in the matrix. Enzymatic degradation remains as the preferred technique as it is inexpensive and eco-friendly. However, the insufficiencies of enzyme battery systems in the degradation of lignocellulosic complex motivate the search for effective degrading enzymes from bacterial isolates from uncommon environment. The study aimed at the evaluation of actinomycetes isolated from saw dust samples collected from wood factory under bed. Cellulase and xylanase production was screened through organism culture on carboxyl methyl cellulose agar and Birchwood xylan. Halo zone indicating lignocellose utilization was shown by an isolate identified through 16S rRNA gene as Micrococcus luteus. The optimum condition for the production of cellulase and xylanase were incubation temperature of 25 °C, fermentation medium pH 5 and 10, agitation speed of 50 and 200 (rpm) and fermentation incubation time of 96 and 84 (h) respectively. The high cellulose and xylanase activity obtained from this isolate portends industrial relevance.

Keywords: carboxyl methyl cellulose, birchwood xylan, optimization, cellulase, xylanase, micrococcus, DNS method

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Authors: Parvin Farzanegi

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Background: Diabetes is a metabolic disorder associated with increased free radicals and oxidative stress. The evidence indicates that physical inactivity is a modifiable behavioral risk factor for a wide range of chronic disorders such as diabetes mellitus. We investigated the effects of eight-week aerobic exercise on some antioxidant enzyme activities in postmenopausal women with type 2 diabetes mellitus (T2DM). Methods: sixteen sedentary postmenopausal women with T2DM were randomly assigned to the control (n=8; CG) and exercise group (n=8; EG). The exercise consisted of progressive aerobic training at a moderate intensity (50-70% of the maximum heart rate), for 25-60 min/day, and 3 days/week for 8 weeks. Age, sex, and body mass index were similar in the two groups. Antioxidant status was evaluated by measuring the superoxide dismutase (SOD) and catalase (CAT) activity. Also levels of malondialdehyde (MDA) as an index of lipid peroxidation and glucose in the plasma were measured before and after the intervention. Results: Following the 8 weeks of exercise training, the plasma MDA and glucose levels were significantly reduced in EG compared to CG (P=0.001 and P=0.011 respectively). However, SOD (P=0.017) and CAT (P=0.011) activities were increased in EG compared to CG. Conclusion: The present study suggests regular aerobic exercise appears can exert protective effects against oxidative stress due to its ability to increase antioxidant defense and glucose control in postmenopausal women with T2DM.

Keywords: aerobic exercise, antioxidant, diabetes mellitus, type 2

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Authors: P. O. Ughachukwu, P. O. Okonkwo, P. C. Unekwe, J. O. Ogamba

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Background: The prevalence of hepatitis B viral infection is high worldwide with liver cirrhosis and hepatocellular carcinoma as important complications. Cases of poor antibody response to hepatitis B vaccination abound. Immunosuppression, especially from glucocorticoids, is often cited as a cause of poor antibody response and there are documented evidences of irrational administration of glucocorticoids to children and adults. The study was, therefore, designed to find out if administration of glucocorticoids affects immune response to vaccination against hepatitis B in mice. Methods: Mice of both sexes were randomly divided into 2 groups. Daily intramuscular methylprednisolone injections, (15 mg kg-1), were given to the test group while sterile deionized water (0.1ml) was given to control mice for 30 days. On day 6 all mice were given 2 μg (0.1ml) hepatitis B vaccine and a booster dose on day 27. On day 34, blood samples were collected and analyzed for anti-HBs titres using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was done using Graph Pad Prism 5.0 and the results taken as statistically significant at p value < 0.05. Results: There were positive serum anti-HBs responses in all mice groups but the differences in titres were not statistically significant. Conclusions: At the dosages and length of exposure used in this study, methylprednisolone injection did not significantly inhibit anti-HBs response in mice following immunization against hepatitis B virus. By extrapolation, methylprednisolone, when used in the usual clinical doses and duration of therapy, is not likely to inhibit immune response to hepatitis B vaccinations in man.

Keywords: anti-HBs, hepatitis B vaccine, immune response, methylprednisolone, mice

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Authors: Mirnawati, G. Ciptaan, Ferawati

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This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.

Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus

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Authors: S. Hallal, Z. Chami, A. Hadji-Lehtihet, S. Sokhal-Boudella, A. Berhoune, L. Yargui

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Gaucher disease is the most common lysosomal storage in our population, it is due to a deficiency of β –glucosidase acid. The enzyme deficiency causes a pathological accumulation of undegraded substrate in lysosomes. This metabolic overload is responsible for a multisystemic disease with hepatosplenomegaly, anemia, thrombocytopenia, and bone involvement. Neurological involvement is rare. The laboratory diagnosis of Gaucher disease consists of phenotypic diagnosis by determining the enzymatic activity of β - glucosidase by fluorimetric method, a study by genotypic diagnosis in the GBA gene, limiting the search recurrent mutations (N370S, L444P, 84 GG); PCR followed by an enzymatic digestion. Abnormal profiles were verified by sequencing. Monitoring of treated patients is provided by the determination of chitotriosidase. Our experience spaning a period of 6 years (2007-2014) has enabled us to diagnose 78 patients out of a total of 328 requests from the various departments of pediatrics, internal medicine, neurology. Genotypic diagnosis focused on the entire family of 9 children treated at pediatric CHU Mustapha, which help define the clinical form; or 5 of them had type III disease, carrying the L444P mutation in the homozygous state. Three others were composite (N370/L444P) (N370S/other unintended mutation in our study), and only in one family no recurrent mutation has been found. This molecular study permits screening of heterozygous essential for genetic counseling.

Keywords: Gaucher disease, mutations, N370S, L444P

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Authors: Naso Isaiah Thanavisuth

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Medical cannabis can be used for treatment including pain, multiple sclerosis, Parkinson's disease, and cancer. However, medical cannabis leads to adverse effects (AEs), which is delta-9-tetrahydrocannabinol (THC). In previous studies, the major of THC metabolism enzymes are CYP2C9. Especially, the variation of CYP2C9 gene consist of CYP2C9*2 on exon 3 and CYP2C9*3 on exon 7 to decrease enzyme activity. Notwithstanding, there is no data describing whether the variant of CYP2C9 genes are apharmacogenetics marker for the prediction of THC-induced AEs in Thai patients. We want to investigate the association between CYP2C9 gene and THC-induced AEs in Thai patients. We enrolled 39 Thai patients with medical cannabis treatment who were classified by clinical data. The CYP2C9*2 and *3 genotyping were conducted using the TaqMan real time PCR assay. All Thai patients who received the medical cannabis consist of twenty-four (61.54%) patients were female, and fifteen (38.46%) were male, with age range 27- 87 years. Moreover, the most AEs in Thai patients who were treated with medical cannabis between cases and controls were tachycardia, arrhythmia, dry mouth, and nausea. Particularly, thirteen (72.22%) medical cannabis-induced AEs were female and age range 33 – 69 years. In this study, none of the medical cannabis groups carried CYP2C9*2 variants in Thai patients. The CYP2C9*3 variants (*1/*3, intermediate metabolizer, IM) and (*3/*3, poor metabolizer, PM) were found, three of thirty-nine (7.69%) and one of thirty-nine (2.56%), respectively. Although, our results indicate that there is no found the CYP2C9*2. However, the variation of CYP2C9 allele might serve as a pharmacogenetics marker for screening before initiating the therapy with medical cannabis for the prevention of medical cannabis-induced AEs.

Keywords: CYP2C9, medical cannabis, adverse effects, THC, P450

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Authors: Hawon Lee, Young-Pil Kim

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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

Procedia PDF Downloads 178

Authors: Mei-Ying Huang, Huei-Jen Ju, Liang-Wei Tseng, Chin-Jung Hsu

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The aim of this study was to investigate a probiotic, Leuconostoc mesenteroides B4, and its products, isomaltooligosaccharide and dextran, on growth performance, digestive enzymes, immune responses, and pathogen resistance of spotted grouper Epinephelus coioides. The grouper were fed control and diets supplemented with L. mesenteroides B4 (107 CFU/g), isomaltooligosaccharide (0.15%), isomaltooligosaccharide (0.15%) + L. mesenteroides B4 (107 CFU/g) (I + B4), and dextran (0.15%) + L. mesenteroides B4 (107 CFU/g) (D + B4) for 8 weeks. The result showed that final weights and percent weight gains of the grouper fed diets supplemented with L. mesenteroides B4 and I + B4 were significantly higher than that of the control group (p < 0.05). The activities of digestive enzymes in the grouper fed with I + B4 were significantly higher than the control group (p < 0.05), too. After challenge with Vibrio harveyi, the enzyme activities of antiprotease and lysozyme as well as of respiratory burst of the fish fed with I + B4 and D + B4 were significantly higher than that of the control group (p < 0.05). The grouper fed with the both diets also had higher survival rates than that of the control group after the challenge. Overall, the study indicated that feeding diets supplemented with L. mesenteroides B4, and its products, isomaltooligosaccharide, and dextran could be an effective method for enhancing the growth performance and disease resistance in orange-spotted grouper.

Keywords: orange-spotted grouper, probiotic Leuconostoc mesenteroides B4, isomaltooligosaccharide, dextran, growth performance, pathogen resistance

Procedia PDF Downloads 239

Authors: Atena Naeimi, Mehri-Sadat Ekrami-Kakhki

Abstract:

Fe3O4 is one of the important magnetic oxides with spinel structure; it has exhibited unique electric and magnetic properties based on the electron transfer between Fe2+ and Fe3+ in the octahedral sites. Fe3O4 have received considerable attention in various areas such as cancer therapy, drug targeting, enzyme immobilization catalysis, magnetic cell separation, magnetic refrigeration systems and super-paramagnetic materials. Fe3O4–ZnO nanostructures were synthesized via a surfactant-free ultrasonic reaction at room temperatures. The effect of various parameters such as temperature, time, and power on the size and morphology of the product was investigated. Alternating gradient force magnetometer shows that Fe3O4 nanoparticles exhibit super-paramagnetic behaviour at room temperature. For preparation of nanocomposite 1 g of Fe3O4 nanostructures were dispersed in 100 mL of distilled water. 0.25 g of Zn (NO3)2 and 20 mL of NH3 solution 1 M were then slowly added to the solution under ultrasonic irradiation. The product was centrifuged, washed with distilled water and dried in the air. The photocatalytic behaviour of Fe3O4–ZnO nanoparticles was evaluated using the degradation of a methyl orange aqueous solution under ultraviolet light irradiation. As time increased, more and more methyl orange was adsorbed on the nanoparticles catalyst, until the absorption peak vanish. The methyl orange concentration decreased rapidly with increasing UV-irradiation time.

Keywords: nanocomposite, ultrasonic, paramagnetic, photocatalytic

Procedia PDF Downloads 279

Authors: Stella O. Olubodun, George E. Eriyamremu

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The study examined the effect of Bonny Light whole crude oil (WC) and its water soluble fraction (WSF) on the activities of antioxidant enzymes (catalase (CAT) and superoxide dismutase (SOD)) and crude mitochondria ATPases in the radicle of germinating bean (Vigna unguiculata). The percentage germination, level of lipid peroxidation, antioxidant enzyme, and mitochondria Ca2+ and Mg2+ ATPase activities were measured in the radicle of bean after 7, 14, and 21 days post germination. Viable bean seeds were planted in soils contaminated with 10ml, 25ml, and 50ml of whole crude oil (WC) and its water soluble fraction (WSF) to obtain 2, 5, and 10% v/w crude oil contamination. There was dose dependent reduction of the number of bean seeds that germinated in the contaminated soils compared with control (p<0.001). The activities of the antioxidant enzymes, as well as, adenosine triphosphatase enzymes, were also significantly (p<0.001) altered in the radicle of the plants grown in contaminated soil compared with the control. Generally, the level of lipid peroxidation was highest after 21 days post germination when compared with control. Stress to germinating bean caused by Bonny Light crude oil or its water soluble fraction resulted in adaptive changes in crude mitochondria ATPases in the radicle.

Keywords: antioxidant enzymes, bonny light crude oil, radicle, mitochondria ATPases

Procedia PDF Downloads 278

Authors: Kshitij Bhardwaj, R.K. Trivedi, Shipra Dixit

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We used biological detoxification method that converts toxic residue waste of Jatropha curcas oil seeds (non edible oil seed) into industrial bio-products and animal feed material. Present study describes the complete degradation of phorbol esters by Aspergillus Niger strain during solid state fermentation (SSF) of deoiled Jatropha curcas seed cake. Phorbol esters were completely degraded in 15 days under the optimized SSF conditions viz deoiled cake 5.0 gm moistened with 5.0 ml distilled water; inoculum 2 ml of overnight grown Aspergillus niger; incubated at 30◦ C, pH 7.0. This method simultaneously induces the production of Protease enzyme by Aspergillus Niger which has high potential to be used in feedstuffs .The maximum Protease activities obtained were 709.16 mg/ml in Jatropha curcas oil seed cake. The protein isolate had small amounts of phorbol esters, phytic acid, and saponin without any lectin. Its minimum and maximum solubility were at pH 4.0&12.0. Water and oil binding capacities were 3.22 g water/g protein and 1.86 ml oil/g protein respectively.Emulsion activity showed high values in a range of basic pH. We concluded that Jatropha Curcas seed cake has a potential to be used as a novel source of functional protein for food or feed applications.

Keywords: solid state fermentation, Jatropha curcas, oil seed cake, phorbol ester

Procedia PDF Downloads 456

Authors: Wighens Ngoie Ilunga, Pamela J. Welz, Olewaseun O. Oyekola, Daniel Ikhu-Omoregbe

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Currently, most sludge from the wastewater treatment plants of edible oil factories is disposed to landfills, but landfill sites are finite and potential sources of environmental pollution. Production of biodiesel from wastewater sludge can contribute to energy production and waste minimization. However, conventional biodiesel production is energy and waste intensive. Generally, biodiesel is produced from the transesterification reaction of oils with alcohol (i.e., Methanol, ethanol) in the presence of a catalyst. Homogeneously catalysed transesterification is the conventional approach for large-scale production of biodiesel as reaction times are relatively short. Nevertheless, homogenous catalysis presents several challenges such as high probability of soap. The current study aimed to reuse wastewater sludge from the edible oil industry as a novel feedstock for both monounsaturated fats and bioethanol for the production of biodiesel. Preliminary results have shown that the fatty acid profile of the oilseed wastewater sludge is favourable for biodiesel production with 48% (w/w) monounsaturated fats and that the residue left after the extraction of fats from the sludge contains sufficient fermentable sugars after steam explosion followed by an enzymatic hydrolysis for the successful production of bioethanol [29% (w/w)] using a commercial strain of Saccharomyces cerevisiae. A novel nano-magnetic catalyst was synthesised from mineral processing alkaline tailings, mainly containing dolomite originating from cupriferous ores using a modified sol-gel. The catalyst elemental chemical compositions and structural properties were characterised by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FTIR) and the BET for the surface area with 14.3 m²/g and 34.1 nm average pore diameter. The mass magnetization of the nano-magnetic catalyst was 170 emu/g. Both the catalytic properties and reusability of the catalyst were investigated. A maximum biodiesel yield of 78% was obtained, which dropped to 52% after the fourth transesterification reaction cycle. The proposed approach has the potential to reduce material costs, energy consumption and water usage associated with conventional biodiesel production technologies. It may also mitigate the impact of conventional biodiesel production on food and land security, while simultaneously reducing waste.

Keywords: biodiesel, bioethanol, edible oil wastewater sludge, nano-magnetism

Procedia PDF Downloads 120

Authors: Akwu N. A., Naidoo Y., Salau V. F., Olofinsan K. A.

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Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is a Southern African medicinal plant indigenously used with other plants for birthing problems. The anti-diabetic properties of the hexane, chloroform, and methanol extracts of Grewia lasiocarpa stem bark were assessed using in vitro α-glucosidase enzyme inhibition assay. The predictive in silico drug-likeness and toxicity properties of the phytocompounds were conducted using the pKCSM, ADMElab, and SwissADME computer-aided online tools. The highest α-glucosidase percentage inhibition was observed in the hexane extract (86.76%, IC50= 0.24 mg/mL), followed by chloroform (63.08%, IC50= 4.87 mg/mL) and methanol (53.22%, IC50= 9.41 mg/mL); while acarbose, the standard anti-diabetic drug was (84.54%, IC50= 1.96 mg/mL). The α-glucosidase assay revealed that the hexane extract exhibited the strongest carbohydrate inhibiting capacity and is a better inhibitor than the standard reference drug-acarbose. The computational studies also affirm the results observed in the in vitroα-glucosidaseassay. Thus, the extracts of G. lasiocarpa may be considered a potential plant-sourced compound for treating type 2 diabetes mellitus. This is the first study on the anti-diabetic properties of Grewia lasiocarpa hexane, chloroform, and methanol extracts using in vitro and in silico models.

Keywords: grewia lasiocarpa, α-glucosidase inhibition, anti-diabetes, ADMET

Procedia PDF Downloads 80

Authors: Mohd Maniruzzaman, Md. Monjurul Alam, Md. Hafezur Rahaman, Anika Amir Mohona

Abstract:

The consumption of water by various industries is increasing day by day, and the wastewaters from them are increasing as well. These wastewaters consist of various kinds of color, dissolved solids, toxic heavy metals, residual chlorine, and other non-degradable organic materials. If these wastewaters are exposed directly to the environment, it will be hazardous for the environment and personal health. So, it is very necessary to treat these wastewaters before exposing into the environment. In this research, we have demonstrated the successful processing and utilization of fully bio-based cellulose micro crystal (CMC) composite for the removal of heavy metals, dyes, and salinity from industrial wastewaters. Banana rachis micro-cellulose were prepared by acid hydrolysis (H₂SO₄) of banana (Musa acuminata L.) rachis fiber, and Bijoypur raw clay were treated by organic solvent tri-ethyl amine. Composites were prepared with varying different composition of banana rachis nano-cellulose and modified Bijoypur (north-east part in Bangladesh) clay. After the successful characterization of cellulose micro crystal (CMC) and modified clay, our targeted filter was fabricated with different composition of cellulose micro crystal and clay in the locally fabricated packing column with 7.5 cm as thickness of composites fraction. Waste-water was collected from local small textile industries containing basic yellow 2 as dye, lead (II) nitrate [Pb(NO₃)₂] and chromium (III) nitrate [Cr(NO₃)₃] as heavy metals and saline water was collected from Khulna to test the efficiency of banana rachis cellulose micro crystal-clay composite for removing the above impurities. The filtering efficiency of wastewater purification was characterized by Fourier transforms infrared spectroscopy (FTIR), X-ray diffraction (X-RD), thermo gravimetric analysis (TGA), atomic absorption spectrometry (AAS), scanning electron microscopy (SEM) analyses. Finally, our all characterizations data are shown with very high expected results for in industrial application of our fabricated filter.

Keywords: banana rachis, bio-based filter, cellulose micro crystal-clay composite, wastewaters, synthetic dyes, heavy metal, water salinity

Procedia PDF Downloads 103

Authors: Hari Bezwada, Michelle Cheon, Ryan Divan, Hannah Escritor, Michelle Kagramian, Isha Korgaonkar, Maya MacAdams, Udgita Pamidigantam, Richard Pilny, Eleanor Race, Angadh Singh, Nathan Zhang, LeeAnn Nguyen, Fina Liotta

Abstract:

Malaria is a deadly disease caused by the Plasmodium parasite, which continues to develop resistance to current antimalarial drugs. In this research project, the effectiveness of numerous thiazole derivatives was explored in inhibiting the PfPKG, a crucial part of the Plasmodium life cycle. This study involved the synthesis of six thiazole-derived amides to inhibit the PfPKG pathway. Nuclear Magnetic Resonance (NMR) spectroscopy and Infrared (IR) spectroscopy were used to characterize these compounds. Furthermore, AutoDocking software was used to predict binding affinities of these thiazole-derived amides in silico. In silico, compound 6 exhibited the highest predicted binding affinity to PfPKG, while compound 5 had the lowest affinity. Compounds 1-4 displayed varying degrees of predicted binding affinity. In-vitro, it was found that compound 4 had the best percent inhibition, while compound 5 had the worst percent inhibition. Overall, all six compounds had weak inhibition (approximately 30-39% at 10 μM), but these results provide a foundation for future drug discovery experiments.

Keywords: Medicinal Chemistry, Malaria, drug discovery, PfPKG, Thiazole, Plasmodium

Procedia PDF Downloads 64