Search results for: M. tuberculosis inhibitors
Commenced in January 2007
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Edition: International
Paper Count: 639

Search results for: M. tuberculosis inhibitors

159 Differential Effects of Parity, Stress and Fluoxetine Treatment on Locomotor Activity and Swimming Behavior in Rats

Authors: Nur Hidayah Kaz Abdul Aziz, Norhalida Hashim, Zurina Hassan

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Peripartum period is a time where women are vulnerable to depression, and stress may further increase the risk of its occurrence. Use of selective serotonin reuptake inhibitors (SSRI) in the treatment of postpartum depression is a common practice. Comparison of antidepressant treatment, however, is rarely studied between gestated and nulliparous animals exposed to stress. This study was aimed to investigate the effect of parity and stress, as well as fluoxetine (an SSRI) treatment after stress exposure on the behavior of rats. Gestating and nulliparous Sprague Dawley rats were either subjected to chronic stressors or left undisturbed throughout the gestation period. After parturition, all stressors were stopped and some of the stressed rats were treated with fluoxetine (10mg/kg). Hence, the final groups formed were: 1. Non-stressed nulliparous rats, 2. Non-stressed dams, 3. Stressed nulliparous rats, 4. Stressed dams, 5. Fluoxetine-treated stressed nulliparous rats, and 6. Fluoxetine-treated stressed dams. Rats were tested in open field test (OFT), novel object recognition test (NOR) and forced swim test (FST) after weaning of pups. Gestational stress significantly reduced the locomotor activity of rats in OFT (p<0.05), while fluoxetine significantly increased the activity in nulliparous rats (p<0.001) but not the dams. While no differences were observed in NOR, stress and parity inhibited the rats from performing swimming behavior in FST. However, climbing and immobile behaviors in FST were found to have no significant differences, although there is a tendency of effect of treatment for immobility parameter (p=0.06) where fluoxetine-treated stressed dams were being the least immobile. In conclusion, the effects of parity and stress, as well as fluoxetine treatment, depended on the type of behavioral test performed.

Keywords: stress, parity, SSRI, behavioral tests

Procedia PDF Downloads 164
158 Osteoprotective Effect of Lawsonia inermis

Authors: Suraj Muke, Vikas Mankumare, Sadhana Sathaye

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Osteoporosis is the most common metabolic bone disease which affects an estimated 25 million people worldwide, leading to 1 million fractures, 40,000 annual deaths and health costs of billions of dollars. It is estimated that about 80% of total osteoporosis patients are women, amongst which majority are above the age of 45 years. Postmenopausal osteoporosis is associated with lack of intestinal calcium absorption, increasing pro-oxidant and inflammatory mediators. Lawsonia inermis is a biennial dicotyledonous herbaceous shrub is reported to possess a high flavonoid, high phenolic and Inhibitors of osteoclastogenesis like Daphneside and Daphnorin. The present study aimed to screen osteoprotective effect of methanolic extract of Lawsonia inermis (LIM) in rat model along with its antioxidant activity. LIM shows phenolic content 146.3Milligrams of Gallic acid equivalent present per gram of extract and 19.8 Milligrams of rutin per gram of extract of Total flavonoid content with IC50 value 42.99μg/ml. bilateral ovariectomized rat model in which Healthy female wistar rats were used for screening. Treatment with LIM was carried out using graded doses of 25mg/kg, 50mg/kg and 100mg/kg for period of 28 days. The negative control group comprised of ovariectomized rats along with saline treatment for four weeks whereas sham operated rats were used as positive control.LIM showed a decrease in bone turnover by preventing loss of urinary calcium and phosphorous moreover it decreased the alkaline phosphatase levels and loss of bone density is prevented by LIM suggesting decrease in osteoclast activity.

Keywords: antioxidant, osteoclast, osteoporosis, ovariectomized

Procedia PDF Downloads 400
157 Linking Metabolism, Pluripotency and Epigenetic Changes during Early Differentiation of Embryonic Stem Cells

Authors: Arieh Moussaieff, Bénédicte Elena-Herrmann, Yaakov Nahmias, Daniel Aberdam

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Differentiation of pluripotent stem cells is a slow process, marked by the gradual loss of pluripotency factors over days in culture. While the first few days of differentiation show minor changes in the cellular transcriptome, intracellular signaling pathways remain largely unknown. Recently, several groups demonstrated that the metabolism of pluripotent mouse and human cells is different from that of somatic cells, showing a marked increase in glycolysis previously identified in cancer as the Warburg effect. Here, we sought to identify the earliest metabolic changes induced at the first hours of differentiation. High-resolution NMR analysis identified 35 metabolites and a distinct, gradual transition in metabolism during early differentiation. Metabolic and transcriptional analyses showed the induction of glycolysis toward acetate and acetyl-coA in pluripotent cells, and an increase in cholesterol biosynthesis during early differentiation. Importantly, this metabolic pathway regulated differentiation of human and mouse embryonic stem cells. Acetate delayed differentiation preventing differentiation-induced histone de-acetylation in a dose-dependent manner. Glycolytic inhibitors upstream of acetate caused differentiation of pluripotent cells, while those downstream delayed differentiation. Our data suggests that a rapid loss of glycolysis in early differentiation down-regulates acetate and acetyl-coA production, causing a loss of histone acetylation and concomitant loss of pluripotency. It demonstrate that pluripotent stem cells utilize a novel metabolism pathway to maintain pluripotency through acetate/acetyl-coA and highlights the important role metabolism plays in pluripotency and early differentiation of stem cells.

Keywords: pluripotency, metabolomics, epigenetics, acetyl-coA

Procedia PDF Downloads 462
156 Re-Engineering of Traditional Indian Wadi into Ready-to-Use High Protein Quality and Fibre Rich Chunk

Authors: Radhika Jain, Sangeeta Goomer

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In the present study an attempt has been made to re-engineer traditional wadi into wholesome ready-to-use cereal-pulse-based chunks rich in protein quality and fibre content. Chunks were made using extrusion-dehydration combination. Two formulations i.e., whole green gram dhal with instant oats and washed green gram dhal with whole oats were formulated. These chunks are versatile in nature as they can be easily incorporated in day-to-day home-made preparations such as pulao, potato curry and kadhi. Cereal-pulse ratio was calculated using NDpCal%. Limiting amino acids such as lysine, tryptophan, methionine, cysteine and threonine were calculated for maximum amino acid profile in cereal-pulse combination. Time-temperature combination for extrusion at 130oC and dehydration at 65oC for 7 hours and 15 minutes were standardized to obtain maximum protein and fibre content. Proximate analysis such as moisture, fat and ash content were analyzed. Protein content of formulation was 62.10% and 68.50% respectively. Fibre content of formulations was 2.99% and 2.45%, respectively. Using a 5-point hedonic scale, consumer preference trials of 102 consumers were conducted and analyzed. Evaluation of chunks prepared in potato curry, kadi and pulao showed preferences for colour 82%, 87%, 86%, texture and consistency 80%, 81%, 88%, flavour and aroma 74%, 82%, 86%, after taste 70%, 75%, 86% and overall acceptability 77%, 75%, 88% respectively. High temperature inactivates antinutritional compounds such as trypsin inhibitors, lectins, saponins etc. Hence, availability of protein content was increased. Developed products were palatable and easy to prepare.

Keywords: extrusion, NDpCal%, protein quality, wadi

Procedia PDF Downloads 218
155 Ameliorative Effect of Martynia annua Linn. on Collagen-Induced Arthritis via Modulating Cytokines and Oxidative Stress in Mice

Authors: Alok Pal Jain, Santram Lodhi

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Martynia annua Linn. (Martyniaccae) is traditionally used in inflammation and applied locally to tuberculosis glands of camel’s neck. The leaves used topically to bites of venomous insects and wounds of domestic animals. Chemical examination of Martynia annua leaves revealed the presence of glycosides, tannins, proteins, phenols and flavonoids. The present study was aimed to evaluate the anti-arthritic activity of methanolic extract of Martynia annua leaves. Methanolic extract of Martynia annua leaves was tested by using in vivo collagen-induced arthritis mouse model to investigate the anti-rheumatoid arthritis activity. In addition, antioxidant effect of methanolic extract was determined by the estimation of antioxidants level in joint tissues. The severity of arthritis was assessed by arthritis score and edema. Levels of cytokines TNF-α and IL-6, in the joint tissue homogenate were measured using ELISA. A high dose (250 mg/kg) of methanolic extract was significantly reduced the degree of inflammation in mice as compared with reference drug. Antioxidants level and malondialdehyde (MDA) in joint tissue homogenate found significantly (p < 0.05) higher. Methanolic extract at dose of 250 mg/kg modulated the cytokines production and suppressed the oxidative stress in the mice with collagen-induced arthritis. This study suggested that Martynia annua might be alternative herbal medicine for the management of rheumatoid arthritis.

Keywords: Martynia annua, collagen, rheumatoid arthritis, antioxidants

Procedia PDF Downloads 288
154 Evaluation of the Safety Status of Beef Meat During Processing at Slaughterhouse in Bouira, Algeria

Authors: A. Ameur Ameur, H. Boukherrouba

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In red meat slaughterhouses a significant number of organs and carcasses were seized because of the presence of lesions of various origins. The objective of this study is to characterize and evaluate the frequency of these lesions in the slaughterhouse of the Wilaya of BOUIRA. On cattle slaughtered in 2646 and inspected 72% of these carcasses have been no seizures against 28% who have undergone at least one entry. 325 lung (44%), 164 livers (22%), 149 hearts (21%) are the main saisis.38 kidneys members (5%), 33 breasts (4%) and 16 whole carcasses (2%) are less seizures parties. The main reasons are the input hydatid cyst for most seized organs such as the lungs (64.5%), livers (51.8%), hearts (23.2%), hydronephrosis for the kidneys (39.4%), and chronic mastitis (54%) for the breasts. Then we recorded second-degree pneumonia (16%) to the lungs, chronic fascioliasis (25%) for livers. A significant difference was observed (p < 0.0001) by sex, race, origin and age of all cattle having been saisie.une a specific input patterns and So pathology was recorded based on race. The local breed presented (75.2%) of hydatid cyst, (95%) and chronic fascioliasis (60%) pyelonephritis, for against the improved breed presented the entire respiratory lesions include pneumonia (64%) the chronic tuberculosis (64%) and mastitis (76%). These results are an important step in the implementation of the concept of risk assessment as the scientific basis of food legislation, by the identification and characterization of macroscopic damage leading withdrawals in meat and to establish the level of inclusion of these injuries within the recommended risk assessment systems (HACCP).

Keywords: slaughterhouses, meat safety, seizure patterns, HACCP

Procedia PDF Downloads 448
153 Performance Evaluation of Next Generation Shale Stabilizer

Authors: N. K. Thakur

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A major proportion of the formations drilled for the production of hydrocarbons consists of clay containing shales. The petroleum industry has hugely investigated the role of clay minerals and their subsequent effect on wellbore stability during the drilling and production of hydrocarbons. It has been found that when the shale formation comes in contact with water-based drilling fluid, the interaction of clay minerals like montmorillonite with infiltrated water leads to hydration of the clay minerals, which causes shale swelling. When shale swelling proceeds further, it may lead to major drilling complications like caving, pipe sticking, which invariably influences wellbore stability, wellbore diameter, the mechanical strength of shale, stress distribution in the wellbore, etc. These problems ultimately lead to an increase in nonproductive time and additional costs during drilling. Several additives are used to prevent shale instability. Among the popular additives used for shale inhibition in drilling muds, ionic liquids and nanoparticles are emerging to be the best additives. The efficiency of the proposed additives will be studied and compared with conventional clay inhibitors like KCl. The main objective is to develop a highly efficient water-based mud for mitigating shale instability and reducing fluid loss which is environmentally friendly and does not alter the formation permeability. The use of nanoparticles has been exploited to enhance the rheological and fluid loss properties in water-based drilling fluid ionic liquid have attracted significant research interest due to its unique thermal stability. It is referred to as ‘green chemical’. The preliminary experimental studies performed are promising. The application of more effective mud additives is always desirable to make the drilling process techno-economically proficient.

Keywords: ionic liquid, shale inhibitor, wellbore stability, unconventional

Procedia PDF Downloads 179
152 New 5’-O- and 6-Substituted Purine Nucleoside Analogs: Synthesis and Cytotoxic Activity on Selected Human Cancer Cell Lines

Authors: Meral Tuncbilek, Duygu Sac, Irem Durmaz, Rengul Cetin Atalay

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Nucleoside analogs are a pharmacologically diverse family that includes cytotoxic compounds, antiviral agents, and immunosuppressive molecules. Purine nucleoside derivatives such as fludarabine, cladribine, and pentostatin are significant drugs used in chemotherapy for the treatment of solid tumors and hematological malignancies. In this study, we synthesized novel purine ribonucleoside analogs containing a 4-(4-substituted phenylsulfonyl) piperazine in the substituent at N6- and O-substituted sulfonyl group at 5’-position as putative cytotoxic agents. The newly obtained compounds were then characterized for their cytotoxicity in human cancer cell lines. The 5’, 6-disubstituted 9-(β-D-ribofuranosyl)purine derivatives (44-67) were readily obtained from commercially available inosine in seven steps in very cost effective synthesis approach. The newly synthesized compounds were first evaluated for their anti-tumor activities against human liver (Huh7), colon (HCT116) and breast (MCF7) carcinoma cell lines. The IC50 values were in micromolar concentrations with 5’, 6-disubstituted purine nucleoside derivatives. Time-dependent IC50 values for each molecule were also calculated in comparison with known cytotoxic agents Camptothecin (CPT), 5-Fluorouracil (5-FU), Cladribine, Pentostatine and Fludarabine. N6-(4-trifluoromethyl phenyl) / N6-(4-bromophenyl) and 5’-O-(4-methoxybenzene sulfonyl) / 5’-O-(benzenesulfonyl) derivatives 54, 64 displayed the best cytotoxic activity with IC50 values of 8.8, 7 µM against MCF7 cell line. The N6-(4-methylphenyl) analog 50 was also very active (IC50= 10.7 μM) against HCT116 cell line. Furthermore, compound 64 had a better cytotoxic activity than the known cell growth inhibitors 5-FU and Fludarabine on Huh7 (1.5 vs 30.7, 29.9 μM for 5-FU and Fludarabine).

Keywords: cytotoxic activity, Huh7, HCT116, MCF7, nucleoside, synthesis

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151 Exploring 1,2,4-Triazine-3(2H)-One Derivatives as Anticancer Agents for Breast Cancer: A QSAR, Molecular Docking, ADMET, and Molecular Dynamics

Authors: Said Belaaouad

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This study aimed to explore the quantitative structure-activity relationship (QSAR) of 1,2,4-Triazine-3(2H)-one derivative as a potential anticancer agent against breast cancer. The electronic descriptors were obtained using the Density Functional Theory (DFT) method, and a multiple linear regression techniques was employed to construct the QSAR model. The model exhibited favorable statistical parameters, including R2=0.849, R2adj=0.656, MSE=0.056, R2test=0.710, and Q2cv=0.542, indicating its reliability. Among the descriptors analyzed, absolute electronegativity (χ), total energy (TE), number of hydrogen bond donors (NHD), water solubility (LogS), and shape coefficient (I) were identified as influential factors. Furthermore, leveraging the validated QSAR model, new derivatives of 1,2,4-Triazine-3(2H)-one were designed, and their activity and pharmacokinetic properties were estimated. Subsequently, molecular docking (MD) and molecular dynamics (MD) simulations were employed to assess the binding affinity of the designed molecules. The Tubulin colchicine binding site, which plays a crucial role in cancer treatment, was chosen as the target protein. Through the simulation trajectory spanning 100 ns, the binding affinity was calculated using the MMPBSA script. As a result, fourteen novel Tubulin-colchicine inhibitors with promising pharmacokinetic characteristics were identified. Overall, this study provides valuable insights into the QSAR of 1,2,4-Triazine-3(2H)-one derivative as potential anticancer agent, along with the design of new compounds and their assessment through molecular docking and dynamics simulations targeting the Tubulin-colchicine binding site.

Keywords: QSAR, molecular docking, ADMET, 1, 2, 4-triazin-3(2H)-ones, breast cancer, anticancer, molecular dynamic simulations, MMPBSA calculation

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150 Induction of G1 Arrest and Apoptosis in Human Cancer Cells by Panaxydol

Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee

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In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.

Keywords: apoptosis, cancer, G1 arrest, panaxydol

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149 Traffic Accident Risk Assessment on National Roads: A Case Study in East Aceh Regency

Authors: Muksalmina

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Transportation plays an important role in people's daily activities but is often marred by traffic accidents. In Indonesia, traffic accidents are the third leading cause of death after coronary heart disease and tuberculosis, according to the World Health Organization (2013). Several roads in East Aceh District are strategic access points for economic growth in the Aceh region. There were 446 traffic accidents in 2023, which is the highest case in the last five years. This study aims to analyze black spot locations on national roads in East Aceh District and evaluate road safety deficiencies in the area. The research methodology began by selecting the locations with the highest accident rates based on data from East Aceh Police from 2019-2023. Next, Average Daily Traffic (ADT) was measured by projecting population growth data. The analysis of road safety deficiencies included measurements of road geometrics, traffic signs and markings, and traffic volumes at black spot locations. The study results showed deficiencies in lane width, shoulder width, and inadequate road safety facilities at several locations. Recommendations for improvements include increasing lane and shoulder widths and adding signs and markings to improve safety. This study is expected to serve as a reference for the government and relevant stakeholders in improving traffic safety in East Aceh District.

Keywords: black spot, traffic accident, severity index, road safety

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148 Sublethal Effect of Tebufenozide, an Ecdysteroid Agonist, on the Reproduction of German Cockroach (Blattodea: Blattellidae)

Authors: Samira Kilani-Morakchi, Amina Badi, Nadia Aribi

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German cockroach, Blattella germanica, is known to be an important pest due to its high reproductive potential and its ability to build up large infectious populations. The infestations were generally controlled by neurotoxic insecticides including organophosphates (OP), carbamate and pyrethroids. An alternative cockroach’s control approach is the use insect growth regulators (IGRs). The relative fewer effects of these chemicals on non-target insects and animals, and their favourable environmental fate, make them attractive insecticides for inclusion in integrated pest management programmes. The juvenoids and chitin synthesis inhibitors are two classes of IGRs that have received the most attention for useful chemicals to manage German cockroaches while ecdysone agonists were mostly used to control Lepidopteran species. In the present study, the sublethal effects of the non-sreroidal ecdysone agonist tebufenozide were evaluated topically on adults of the B. germanica. The effects on reproduction were observed in adults females of cockroaches that survived exposure to LD25 (146 µg/g of insect) of tebufenozide. Dissection of treated females showed a clear reduction in both the number of oocytes per paired ovaries and the size of basal oocytes, as compared to controls. In addition, tebufenozide significantly reduced the mating success of pairs and altered the fertility as shown through the reduction of ootheca development and total absence of viable nymph. Tebufenozide disrupted the German cockroach reproduction by interfering with homeostasis of the insect hormones. In conclusion, the overall results suggested that tebufenozide can be used as a biorational insecticide for controlling cockroaches.

Keywords: B. germanica, ecdysteroid agonist, tebufenozide, reproduction

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147 Eudesmane-Type Sesquiterpenes from Laggera alata Inhibiting Angiogenesis

Authors: Liang Ning, Chung Hau Yin

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Angiogenesis is the process of new blood vessel development. It has been recognized as a therapeutic target for blocking cancer growth four decades ago. Vascular sprouting is initiated by pro-angiogenic factors. Vascular endothelial cell growth factor (VEGF) plays a central role in angiogenic initiation, many patients with cancer or ocular neovascularization have been benefited from anti-VEGF therapy. Emerging approaches impacting in the later stages of vessel remodeling and maturation are expected to improve clinical efficacy. TIE receptor as well as the corresponding angiopoietin ligands, were identified as another endothelial cell specific receptor tyrosine kinase signaling system. Much efforts were made to reduce the activity of angiopoietin-TIE receptor axis. Two eudesmane-type sesquiterpenes from laggera alata, namely, 15-dihydrocostic acid and ilicic acid were found with strong anti-angiogenic properties in zebrafish model. Meanwhile, the mRNA expression levels of VEGFR2 and TIE2 pathway related genes were down-regulated in the sesquiterpenes treated zebrafish embryos. Besides, in human umbilical vein endothelial cells (HUVECs), the sesquiterpenes have the ability to inhibit VEGF-induced HUVECs proliferation and migration at non-toxic concentration. Moreover, angiopoietin-2 induced TIE2 phosphorylation was inhibited by the sesquiterpenes, the inhibitory effect was detected in angiopoietin-1 induced HUVECs proliferation as well. Thus, we hypothesized the anti-angiogenic activity of the compounds may via the inhibition of VEGF and TIE2 related pathways. How the compounds come into play as the pathways inhibitors need to be evaluated in the future.

Keywords: Laggera alata, eudesmane-type sesquiterpene, anti-angiogenesis, VEGF, angiopoietin, TIE2

Procedia PDF Downloads 195
146 An In-Depth Experimental Study of Wax Deposition in Pipelines

Authors: Arias M. L., D’Adamo J., Novosad M. N., Raffo P. A., Burbridge H. P., Artana G.

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Shale oils are highly paraffinic and, consequently, can create wax deposits that foul pipelines during transportation. Several factors must be considered when designing pipelines or treatment programs that prevents wax deposition: including chemical species in crude oils, flowrates, pipes diameters and temperature. This paper describes the wax deposition study carried out within the framework of Y-TEC's flow assurance projects, as part of the process to achieve a better understanding on wax deposition issues. Laboratory experiments were performed on a medium size, 1 inch diameter, wax deposition loop of 15 mts long equipped with a solid detector system, online microscope to visualize crystals, temperature and pressure sensors along the loop pipe. A baseline test was performed with diesel with no paraffin or additive content. Tests were undertaken with different temperatures of circulating and cooling fluid at different flow conditions. Then, a solution formed with a paraffin added to the diesel was considered. Tests varying flowrate and cooling rate were again run. Viscosity, density, WAT (Wax Appearance Temperature) with DSC (Differential Scanning Calorimetry), pour point and cold finger measurements were carried out to determine physical properties of the working fluids. The results obtained in the loop were analyzed through momentum balance and heat transfer models. To determine possible paraffin deposition scenarios temperature and pressure loop output signals were studied. They were compared with WAT static laboratory methods. Finally, we scrutinized the effect of adding a chemical inhibitor to the working fluid on the dynamics of the process of wax deposition in the loop.

Keywords: paraffin desposition, flow assurance, chemical inhibitors, flow loop

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145 Biodiversity Affects Bovine Tuberculosis (bTB) Risk in Ethiopian Cattle: Prospects for Infectious Disease Control

Authors: Sintayehu W. Dejene, Ignas M. A. Heitkönig, Herbert H. T. Prins, Zewdu K. Tessema, Willem F. de Boer

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Current theories on diversity-disease relationships describe host species diversity and species identity as important factors influencing disease risk, either diluting or amplifying disease prevalence in a community. Whereas the simple term ‘diversity’ embodies a set of animal community characteristics, it is not clear how different measures of species diversity are correlated with disease risk. We, therefore, tested the effects of species richness, Pielou’s evenness and Shannon’s diversity on bTB risk in cattle in the Afar Region and Awash National Park between November 2013 and April 2015. We also analysed the identity effect of a particular species and the effect of host habitat use overlap on bTB risk. We used the comparative intradermal tuberculin test to assess the number of bTB infected cattle. Our results suggested a dilution effect through species evenness. We found that the identity effect of greater kudu - a maintenance host – confounded the dilution effect of species diversity on bTB risk. bTB infection was positively correlated with habitat use overlap between greater kudu and cattle. Different diversity indices have to be considered together for assessing diversity-disease relationships, for understanding the underlying causal mechanisms. We posit that unpacking diversity metrics is also relevant for formulating control strategies to manage cattle in ecosystems characterized by seasonally limited resources and intense wildlife-livestock interactions.

Keywords: evenness, diversity, greater kudu, identity effect, maintenance hosts, multi-host disease ecology, habitat use overlap

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144 Preparation of Flurbiprofen Derivative for Enhanced Brain Penetration

Authors: Jungkyun Im

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Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective for relieving pain and reducing inflammation. They are nonselective inhibitors of two isoforms of COX, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), and thereby inhibiting the production of hormone-like lipid compounds such as, prostaglandins and thromboxanes which cause inflammation, pain, fever, platelet aggregation, etc. In addition, recently there are many research articles reporting the neuroprotective effect of NSAIDs in neurodegenerative diseases, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). However, the clinical use of NSAIDs in these diseases is limited by low brain distribution. Therefore, in order to assist the in-depth investigation on the pharmaceutical mechanism of flurbiprofen in neuroprotection and to make flurbiprofen a more potent drug to prevent or alleviate neurodegenerative diseases, delivery of flurbiprofen to brain should be effective and sufficient amount of flurbiprofen must penetrate the BBB thus gaining access into the patient’s brain. We have recently developed several types of guanidine-rich molecular carriers with high molecular weights and good water solubility that readily cross the blood-brain barrier (BBB) and display efficient distributions in the mouse brain. The G8 (having eight guanidine groups) molecular carrier based on D-sorbitol was found to be very effective in delivering anticancer drugs to a mouse brain. In the present study, employing the same molecular carrier, we prepared the flurbiprofen conjugate and studied its BBB permeation by mouse tissue distribution study. Flurbiprofen was attached to a molecular carrier with a fluorescein probe and multiple terminal guanidiniums. The conjugate was found to internalize into live cells and readily cross the BBB to enter the mouse brain. Our novel synthetic flurbiprofen conjugate will hopefully delivery NSAIDs into brain, and is therefore applicable to the neurodegenerative diseases treatment or prevention.

Keywords: flurbiprofen, drug delivery, molecular carrier, organic synthesis

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143 Role of Basic Health Units in Provision of Primary Health Services in District Swabi

Authors: Naila Awan, Shahrukh Inam

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This study was conducted to highlight the role of basic health units in district Swabi, which provides primary health services to the people of district Swabi having four tehsils. Tehsil Swabi was selected purposively for the study. Three villages were purposively selected from district Swabi. A sum of 110 respondents was randomly selected for interview i.e., 27 from Botakaa, 39 from Gulatee, and 44 from Darra Cham, using proportion allocation sampling technique. A pretested and well-designed interview schedule was used to collect as per the objective and Chi square test was applied to find an association between the quality of medicines and health improvement. The output of the test shows that the government was doing its best and providing enough facilities to the individuals at the healthcare units, and they were utilizing them. These resources were easily accessible to the people of the community. Medicines provided by the government were of good quality and quantity. There were also school health sessions and community health sessions (SHS/CHS) to deliver useful information and awareness regarding health problems and diseases were conducted. The staff of the BHU was present at work time and was performing their duties. The respondents seemed satisfied with their behavior and the duty of the staff. However, there were no emergency resources existing at the BHU after the working hours of the medical staff. It is recommended that government should provide appropriate quantity and quality of medicines to the basic health units so that these healthcare units don’t have to face any shortages regarding medicines at the end of the month. In addition, laboratory and blood testing facilities need to be provided in the basic health units, and also the infrastructure should be made suitable, satisfactory, and more functional.

Keywords: community health session, basic health units, outpatient department, tuberculosis

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142 Anti-TNF: Possibilities of Rising Anti-Phosphorylcholine Antibodies

Authors: Md. Mizanur Rahman, Anquan Liu, Anna Frostegård, Johan Frostegård

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The role of the human immune system is essential in cardiovascular diseases and atherosclerosis. Activated cells in atherosclerosis produce abundant amounts of cytokines, but the exact mechanisms involved in the effects of these inflammatory cytokines are not clear in atherosclerosis. In a large clinical cohort, we have previously determined that antibodies against phosphorylcholine (anti-PC) are negatively and independently associated with both development of atherosclerosis and also a low risk of cardiovascular disease. Further, we reported that rheumatoid arthritis patients who were non-responders to TNF-inhibitors, where those with low anti-PC levels. Upon anti-TNF treatment, anti-PC levels increased. We, therefore, hypothesised that proinflammatory cytokines such as TNF could play a role in anti-PC regulation. Peripheral blood mononuclear cells (PBMC) were cultured with or without TNF and anti-TNF. The cell supernatants were collected after six days for ELISA measurements. In separate experiments, cells were cultured for 24 hours in both polystyrene plates and ELISPOT plates under a similar condition for ELISA and ELISPOT assays respectively. Total RNA was extracted after 6 hours of cell culture to perform RT-qPCR. Cell viability was confirmed by trypan blue staining and MTT assays. ELISA measurements detected less than 40% of anti-PC in TNF-treated cells, in comparison to control cells, whereas anti-PC production was recovered by anti-TNF treatment. ELISPOT assays showed that TNF suppresses anti-PC production by inhibiting anti-PC producing B-cells. In addition, RT-qPCR and ELISA showed that TNF also has effects also on B-cell activation as BAFF expression was inhibited by TNF treatment. Atherosclerosis is a major cause of cardiovascular diseases, but anti-PC is a protection marker for atherosclerosis development. Our findings show that TNF is a negative regulator of anti-PC production. Immune modulation and rising of anti-PC could be of major significance for the patients.

Keywords: anti-PC, Anti-TNF, atherosclerosis, cardiovascular diseases, phosphorylecholine

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141 Susceptibility of Spodoptera littoralis, Field Populations in Egypt to Chlorantraniliprole and the Role of Detoxification Enzymes

Authors: Mohamed H. Khalifa, Fikry I. El-Shahawi, Nabil A. Mansour

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The cotton leafworm, Spodoptera littoralis (Boisduval) is a major insect pest of vegetables and cotton crops in Egypt, and exhibits different levels of tolerance to certain insecticides. Chlorantraniliprole has been registered recently in Egypt for control this insect. The susceptibilities of three S. littoralis populations collected from El Behaira governorate, north Egypt to chlorantraniliprole were determined by leaf-dipping technique on 4th instar larvae. Obvious variation of toxicity was observed among the laboratory susceptible, and three field populations with LC50 values ranged between 1.53 µg/ml and 6.22 µg/ml. However, all the three field populations were less susceptible to chlorantraniliprole than a laboratory susceptible population. The most tolerant populations were sampled from El Delengat (ED) Province where S. littoralis had been frequently challenged by insecticides. Certain enzyme activity assays were carried out to be correlated with the mechanism of the observed field population tolerance. All field populations showed significantly enhanced activities of detoxification enzymes compared with the susceptible strain. The regression analysis between chlorantraniliprole toxicities and enzyme activities revealed that the highest correlation is between α-esterase or β-esterase (α-β-EST) activity and collected field strains susceptibility, otherwise this correlation is not significant (P > 0.05). Synergism assays showed the ED and susceptible strains could be synergized by known detoxification inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.01-8.76-fold and 1.09-2.94 fold, respectively), TPP showed the maximum synergism in both strains. The results show that there is a correlation between the enzyme activity and tolerance, and carboxylic-esterase (Car-EST) is likely the main detoxification mechanism responsible for tolerance of S. littoralis to chlorantraniliprole.

Keywords: chlorantraniliprole, detoxification enzymes, Egypt, Spodoptera littoralis

Procedia PDF Downloads 267
140 The Impact of Pediatric Cares, Infections and Vaccines on Community and People’s Lives

Authors: Nashed Atef Nashed Farag

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Introduction: Reporting adverse events following vaccination remains a challenge. WHO has mandated pharmacovigilance centers around the world to submit Adverse Events Following Immunization (AEFI) reports from different countries to a large electronic database of adverse drug event data called Vigibase. Despite sufficient information about AEFIs on Vigibase, they are not available to the general public. However, the WHO has an alternative website called VigiAccess, an open-access website that serves as an archive for reported adverse reactions and AEFIs. The aim of the study was to establish a reporting model for a number of commonly used vaccines in the VigiAccess system. Methods: On February 5, 2018, VigiAccess comprehensively searched for ESSI reports on the measles vaccine, oral polio vaccine (OPV), yellow fever vaccine, pneumococcal vaccine, rotavirus vaccine, meningococcal vaccine, tetanus vaccine, and tuberculosis vaccine (BCG). These are reports from all pharmacovigilance centers around the world since they joined the WHO Drug Monitoring Program. Results: After an extensive search, VigiAccess found 9,062 AEFIs from the measles vaccine, 185,829 AEFIs from the OPV vaccine, 24,577 AEFIs from the yellow fever vaccine, 317,208 AEFIs from the pneumococcal vaccine, 73,513 AEFIs from the rotavirus vaccine, and 145,447 AEFIs from meningococcal cal vaccine, 22,781 EI FI vaccines against tetanus and 35,556 BCG vaccines against AEFI. Conclusion: The study found that among the eight vaccines examined, pneumococcal vaccines were associated with the highest number of AEFIs, while measles vaccines were associated with the fewest AEFIs.

Keywords: surgical approach, anatomical approach, decompression, axillary nerve, quadrangular space adverse events following immunization, cameroon, COVID-19 vaccines, nOPV, ODK vaccines, adverse reactions, VigiAccess, adverse event reporting

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139 Availability of TB Infection Control Plans at Rural Hospitals of South Africa

Authors: Takalani Tshitangano

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Background: In Limpopo province the rate of new tuberculosis (TB) cases increase daily. The Infection Control (IC) plan is one of the essential actions for TB IC. This study aimed to establish the availability of these plans at health care facilities. Objectives: The objectives were to explore and describe the awareness and knowledge of health care workers (HCWs) of the availability and content of TB IC plan; and to identity the role of infection control committees from the perspective of HCWs. Method: A qualitative approach using a cross-sectional descriptive design was adopted. The target population was all HCWs from the seven hospitals of Vhembe district. A purposive sampling approach was used to select 57 participants. The approval to conduct this study was obtained from the relevant authorities and participants. Data were collected through seven focus group discussions comprising five to 10 members. An unstructured discussion guide was used to collect data, and an open-coding method was used to analyse the data. Lincoln and Guba’s criteria ensured trustworthiness of the study findings. Results: Findings revealed that HCWs were not aware of the availability and the information contained in the TB IC plans. No person was designated as TB IC officer at hospital level. There was lack of a TB IC Committee and teams as well as ineffective utilisation of those that did exist. Conclusions: It was concluded that if the TB IC plans are not available at health care facilities, then the TB IC practices implemented by HCWs vary, resulting in TB nosocomial infection transmission. It was recommended that the World Health Organisation’s TB IC plans be adopted and implemented in Vhembe district.

Keywords: health care workers' awareness, health care workers' knowledge, availability of TB infection control plans, rural hospitals

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138 Improvement of Microscopic Detection of Acid-Fast Bacilli for Tuberculosis by Artificial Intelligence-Assisted Microscopic Platform and Medical Image Recognition System

Authors: Hsiao-Chuan Huang, King-Lung Kuo, Mei-Hsin Lo, Hsiao-Yun Chou, Yusen Lin

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The most robust and economical method for laboratory diagnosis of TB is to identify mycobacterial bacilli (AFB) under acid-fast staining despite its disadvantages of low sensitivity and labor-intensive. Though digital pathology becomes popular in medicine, an automated microscopic system for microbiology is still not available. A new AI-assisted automated microscopic system, consisting of a microscopic scanner and recognition program powered by big data and deep learning, may significantly increase the sensitivity of TB smear microscopy. Thus, the objective is to evaluate such an automatic system for the identification of AFB. A total of 5,930 smears was enrolled for this study. An intelligent microscope system (TB-Scan, Wellgen Medical, Taiwan) was used for microscopic image scanning and AFB detection. 272 AFB smears were used for transfer learning to increase the accuracy. Referee medical technicians were used as Gold Standard for result discrepancy. Results showed that, under a total of 1726 AFB smears, the automated system's accuracy, sensitivity and specificity were 95.6% (1,650/1,726), 87.7% (57/65), and 95.9% (1,593/1,661), respectively. Compared to culture, the sensitivity for human technicians was only 33.8% (38/142); however, the automated system can achieve 74.6% (106/142), which is significantly higher than human technicians, and this is the first of such an automated microscope system for TB smear testing in a controlled trial. This automated system could achieve higher TB smear sensitivity and laboratory efficiency and may complement molecular methods (eg. GeneXpert) to reduce the total cost for TB control. Furthermore, such an automated system is capable of remote access by the internet and can be deployed in the area with limited medical resources.

Keywords: TB smears, automated microscope, artificial intelligence, medical imaging

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137 Mechanism of Modeling the Level of Bcr-Abl Oncoprotein by Ubiquitin-Proteasome System in Chronic Myeloid Leukemia

Authors: Svitlana Antonenko, Gennady Telegeev

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Introductive statement: The development of chronic myeloid leukemia (CML) is caused by Bcr-Abl oncoprotein. Modern treatments with tyrosine kinase inhibitors are greatly complicated by the mutational variability of the Bcr-Abl oncoprotein, which causes drug resistance. Therefore, there is an urgent need to develop new approaches to the treatment of the disease, which will allow modeling the level of Bcr-Abl oncoprotein in the cell. Promising in this direction is the identification of proteases that can selectively promote cellular proteolysis of oncoproteins. The aim of the study was to study the effect of the interaction of Bcr-Abl with deubiquitinase USP1 on the level of oncoprotein in CML cells. Methodology: K562 cells were selected for the experiment. Сells were incubated with ML323 inhibitor for 24 hours. Precipitation of endogenous proteins from K562 cell lysate was performed using anti-Bcr-Abl antibodies. Cell lysates and precipitation results were studied by Western blot. Subcellular localization of proteins was studied by immunofluorescence analysis followed by confocal microscopy. The results were analyzed quantitatively and statistically. Major findings: The Bcr-Abl/USP1 protein complex was detected in CML cells, and it was found that inhibition of USP1 deubiquitinating activity by the compound ML323 leads to disruption of this protein complex and a decrease in the level of Bcr-Abl oncoprotein in cells. The interaction of Bcr-Abl with USP1 may result in deubiquitination of the oncoprotein, which disrupts its proteasomal degradation and leads to the accumulation of CML in cells. Conclusion: We believe that the interaction of oncoprotein with USP1 may be one of the prerequisites that contribute to malignant cell transformation due to the deubiquitination of oncoprotein, which leads to its accumulation and disease progression. A correlation was found between the deubiquitinating activity of USP1 and the level of oncoprotein in CML cells. Thus, we identify deubiquitinase USP1 as a promising therapeutic target for the development of a new strategy for the treatment of CML by modulating the level of Bcr-Abl in the cell.

Keywords: chronic myeloid leukemia, Bcr-Abl, USP1, deubiquitination Bcr-Abl, K562 cell

Procedia PDF Downloads 61
136 Synthesis and Biological Activities of Novel -1,2,3-Triazoles Derivatives

Authors: Zahra Dehghani, Hoda Dehghani, Elham Zarenezhad

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1,2,3-Triazole derivatives are important compounds in medicinal chemistry owing to their wide applications in drug discovery. They can readily associate with biologically targets through the hydrogen bonding and dipole interactions. The 1,2,3-triazole core is a key structural motif in many bioactive compounds, exhibiting a broad spectrum of biological activities, such as antiviral, anticancer, anti-HIV, antibiotic, antibacterial, and antimicrobial. Additionally, they have found significant industrial applications as dyes, agrochemicals, corrosion inhibitors, photo stabilizers, and photographic materials. we disclose the synthesis and characterization of 1-azido-3-(aryl-2-yloxy)propan-2-ol drivatives. The chemistry works well with various ß-azido alcohols involving aryloxy, alkoxy and alkyl residues, and also tolerates a wide spectrum of electron-donating and electron-withdrawing functional groups in both alkyne and azide molecules. Most of ß-azidoalcohols used in these experiments were pre-synthesized by the regioselective ring opening reaction of corresponded epoxides with sodium azide, whereas the majority of terminal alkynes were prepared via SN2-type reaction of propargyl bromide and corresponded nucleophiles. To evaluate the bioactivity of title compounds, the in vitro antifungal activity of all compound was investigated against several pathogenic fungi including Candida albicans, Candida krusei, Aspergillus niger, and Trichophyton rubrum , clotrimazole and fluconazole was used as standard antifungal drugs, also To understand the antibacterial activity of synthesized compounds, they were in vitro screened against E. coli and S. aureus as Gram-negative and Gram-positive bacteria, respectively. The in vitro tests have shown the promising antifungal but marginal antibacterial activity against tested fungi and bacteria.

Keywords: biological activities, antibacterial, antifungal, 1, 2, 3-Triazole

Procedia PDF Downloads 421
135 Combined Treatment of Aged Rats with Donepezil and the Gingko Extract EGb 761® Enhances Learning and Memory Superiorly to Monotherapy

Authors: Linda Blümel, Bettina Bert, Jan Brosda, Heidrun Fink, Melanie Hamann

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Age-related cognitive decline can eventually lead to dementia, the most common mental illness in elderly people and an immense challenge for patients, their families and caregivers. Cholinesterase inhibitors constitute the most commonly used antidementia prescription medication. The standardized Ginkgo biloba leaf extract EGb 761® is approved for treating age-associated cognitive impairment and has been shown to improve the quality of life in patients suffering from mild dementia. A clinical trial with 96 Alzheimer´s disease patients indicated that the combined treatment with donepezil and EGb 761® had fewer side effects than donepezil alone. In an animal model of cognitive aging, we compared the effect of combined treatment with EGb 761® or donepezil monotherapy and vehicle. We compared the effect of chronic treatment (15 days of pretreatment) with donepezil (1.5 mg/kg p. o.), EGb 761® (100 mg/kg p. o.), or the combination of the two drugs, or vehicle in 18 – 20 month old male OFA rats. Learning and memory performance were assessed by Morris water maze testing, motor behavior in an open field paradigm. In addition to chronic treatment, the substances were administered orally 30 minutes before testing. Compared to the first day and to the control group, only the combination group showed a significant reduction in latency to reach the hidden platform on the second day of testing. Moreover, from the second day of testing onwards, the donepezil, the EGb 761® and the combination group required less time to reach the hidden platform compared to the first day. The control group did not reach the same latency reduction until day three. There were no effects on motor behavior. These results suggest a superiority of the combined treatment of donepezil with EGb 761® compared to monotherapy.

Keywords: age-related cognitive decline, dementia, ginkgo biloba leaf extract EGb 761®, learning and memory, old rats

Procedia PDF Downloads 358
134 Associations between Physical Activity and Risk Factors for Type II Diabetes in Prediabetic Adults

Authors: Rukia Yosuf

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Diabetes is a national healthcare crisis related to both macrovascular and microvascular complications. We hypothesized that higher levels of physical activity are associated with lower total and visceral fat mass, lower systolic blood pressure, and increased insulin sensitivity. Participant inclusion criteria: 21-50 years old, BMI ≥ 30 kg/m2, hemoglobin A1C 5.7-6.4, fasting glucose 100-125 mg/dL, and HOMA IR ≥ 2.5. Exclusion criteria: history of diabetes, hypertension, HIV, renal disease, hearing loss, alcoholic intake over four drinks daily, use of organic nitrates or PDE5 inhibitors, and decreased cardiac function. Total physical activity was measured using accelerometers, body composition using DXA, and insulin resistance via fsIVGTT. Clinical and biochemical cardiometabolic risk factors, blood pressure and heart rate were obtained using a calibrated sphygmomanometer. Anthropometric measures, fasting glucose, insulin, lipid profile, C-reactive protein, and BMP were analyzed using standard procedures. Within our study, we found correlations between levels of physical activity in a heterogeneous group of prediabetic adults. Patients with more physical activity had a higher degree of insulin sensitivity, lower blood pressure, total visceral adipose tissue, and overall lower total mass. Total physical activity levels showed small, but significant correlations with systolic blood pressure, visceral fat, lean mass and insulin sensitivity. After normalizing for the race, age, and gender using multiple regression, these associations were no longer significant considering our small sample size. More research into prediabetes will decrease the population of diabetics overall. In the future, we could increase sample size and conduct cross sectional and longitudinal studies in various populations with prediabetes.

Keywords: diabetes, kidney disease, nephrology, prediabetes

Procedia PDF Downloads 180
133 Development and Validation Method for Quantitative Determination of Rifampicin in Human Plasma and Its Application in Bioequivalence Test

Authors: Endang Lukitaningsih, Fathul Jannah, Arief R. Hakim, Ratna D. Puspita, Zullies Ikawati

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Rifampicin is a semisynthetic antibiotic derivative of rifamycin B produced by Streptomyces mediterranei. RIF has been used worldwide as first line drug-prescribed throughout tuberculosis therapy. This study aims to develop and to validate an HPLC method couple with a UV detection for determination of rifampicin in spiked human plasma and its application for bioequivalence study. The chromatographic separation was achieved on an RP-C18 column (LachromHitachi, 250 x 4.6 mm., 5μm), utilizing a mobile phase of phosphate buffer/acetonitrile (55:45, v/v, pH 6.8 ± 0.1) at a flow of 1.5 mL/min. Detection was carried out at 337 nm by using spectrophotometer. The developed method was statistically validated for the linearity, accuracy, limit of detection, limit of quantitation, precise and specifity. The specifity of the method was ascertained by comparing chromatograms of blank plasma and plasma containing rifampicin; the matrix and rifampicin were well separated. The limit of detection and limit of quantification were 0.7 µg/mL and 2.3 µg/mL, respectively. The regression curve of standard was linear (r > 0.999) over a range concentration of 20.0 – 100.0 µg/mL. The mean recovery of the method was 96.68 ± 8.06 %. Both intraday and interday precision data showed reproducibility (R.S.D. 2.98% and 1.13 %, respectively). Therefore, the method can be used for routine analysis of rifampicin in human plasma and in bioequivalence study. The validated method was successfully applied in pharmacokinetic and bioequivalence study of rifampicin tablet in a limited number of subjects (under an Ethical Clearance No. KE/FK/6201/EC/2015). The mean values of Cmax, Tmax, AUC(0-24) and AUC(o-∞) for the test formulation of rifampicin were 5.81 ± 0.88 µg/mL, 1.25 hour, 29.16 ± 4.05 µg/mL. h. and 29.41 ± 4.07 µg/mL. h., respectively. Meanwhile for the reference formulation, the values were 5.04 ± 0.54 µg/mL, 1.31 hour, 27.20 ± 3.98 µg/mL.h. and 27.49 ± 4.01 µg/mL.h. From bioequivalence study, the 90% CIs for the test formulation/reference formulation ratio for the logarithmic transformations of Cmax and AUC(0-24) were 97.96-129.48% and 99.13-120.02%, respectively. According to the bioequivamence test guidelines of the European Commission-European Medicines Agency, it can be concluded that the test formulation of rifampicin is bioequivalence with the reference formulation.

Keywords: validation, HPLC, plasma, bioequivalence

Procedia PDF Downloads 284
132 Investigating the Flavin-Dependent Thymidylate Synthase (FDTS) Enzyme from Clostridioides Difficile (C. diff)

Authors: Sidra Shaw, Sarenna Shaw, Chae Joon Lee, Irimpan Mathews, Eric Koehn

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One of the biggest public health concerns of our time is increasing antimicrobial resistance. As of 2019, the CDC has documented more than 2.8 million serious antibiotic resistant infections in the United States. Currently, antibiotic resistant infections are directly implicated in over 750,000 deaths per year globally. On our current trajectory, British economist Jim O’Neill predicts that by 2050, an additional 10 million people (about half the population of New York) will die annually due to drug resistant infections. As a result, new biochemical pathways must be targeted to generate next generation antibiotic drugs that will be effective against drug resistant bacteria. One enticing target is the biosynthesis of DNA within bacteria, as few drugs interrupt this essential life process. Thymidylate synthase enzymes are essential for life as they catalyze the synthesis of a DNA building block, 2′-deoxythymidine-5′-monophosphate (dTMP). In humans, the thymidylate synthase enzyme (TSase) has been shown to be distinct from the flavin-dependent thymidylate synthase (FDTS) produced by many pathogenic bacteria. TSase and FDTS have distinct structures and mechanisms of catalysis, which should allow selective inhibition of FDTS over human TSase. Currently, C. diff is one of the most antibiotic resistant bacteria, and no drugs that target thymine biosynthesis exist for C. diff. Here we present the initial biochemical characterization of FDTS from C. diff. Specifically, we examine enzyme kinetics and binding features of this enzyme to determine the nature of interaction with ligands/inhibitors and understand the molecular mechanism of catalysis. This research will provide more insight into the targetability of the C. diff FDTS enzyme for novel antibiotic drugs.

Keywords: flavin-dependent thymidylate synthase, FDTS, clostridioides difficile, C. diff, antibiotic resistance, DNA synthesis, enzyme kinetics, binding features

Procedia PDF Downloads 91
131 Identification and Application of Biocontrol Agents against Cotton Leaf Curl Virus Disease in Gossypium hirsutum under Green House Conditions

Authors: Memoona Ramzan, Bushra Tabassum, Anwar Khan, Muhammad Tariq, Mudassar Fareed Awan, Idrees Ahmad Nasir, Zahida Qamar, Naila Shahid, Tayyab Husnain

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Biological control is a novel approach being used in crop protection nowadays. Bacteria like Bacillus and Pseudomonas are reported for this purpose and few of their products are commercially available too. Rhizosphere and phyllosphere of healthy cotton plants were used as a source to isolate bacteria capable of exhibiting properties worthy for selection as biocontrol agent. For this purpose all isolated strains were screened for the activities like phosphate solubilization, Indole acetic acid (IAA) production and biocontrol against fungi. Two strains S1HL3 and S1HL4 showed phosphate solubilization and IAA production simultaneously while two other JS2HR4 and JS3HR2 were good inhibitors of fungal pathogens. Through biochemical and molecular characterization these bacteria were identified as P. aeruginosa, Burkholderia and Bacillus respectively. In green house trials of these isolates against Cotton leaf curl virus (CLCuV), seven treatments including individual bacterial isolate and consortia were included. Treated plants were healthy as compared to control plants in which upto 74% CLCuV symptomatic plants exist. Maximum inhibition of CLCuV was observed in T7 treated plants where viral load was only 0.4% as compared to control where viral load was upto 74%. This treatment consortium included Bacillus and Pseudomonas isolates; S1HL3, S1HL4, JS2HR4 and JS3HR2. Principal Component Biplot depicted highly significant correlation between percentage viral load and the disease incidence.

Keywords: cotton leaf curl virus, biological control, bacillus, pseudomonas

Procedia PDF Downloads 374
130 Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes

Authors: A. K. R. Gobinath, He Jianzhong, Kun-Lin Yang

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Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.

Keywords: autoclave pretreatment, biohydrogen production, clostridial fermentation, fruit waste, and microwave pretreatment

Procedia PDF Downloads 524