Search results for: gas hydrate promoter

Authors: Pillay J. D., D. De Wit, J. F. Ducray

Abstract:

Field hockey is a popular international sport which is played in more than 100 countries across the world. Due to the nature of hockey, players repeatedly perform a combination of forward flexion and rotational movements of the spine in order to strike the ball. These movements have been shown to increase the risk of pain and injury to the lumbar spine. The aim of this study was to determine the prevalence and incidence of low back pain (LBP) in male adolescent field hockey players and the characteristics of LBP in terms of location, chronicity, disability, and treatment sought, as well as its association with selected risk factors. A survey was conducted on 112 male adolescent field hockey players in the eThekwini Municipality of KwaZulu-Natal, South Africa. The questionnaire contained sections on the demographics of participants, general characteristics of participants, health and lifestyle characteristics, low back pain patterns, treatment of low back pain, and the level of disability associated with LBP. The data were statistically analysed using IBM SPSS version 25 with statistical significance set at p-value <0.05. Descriptive statistics such as mean and standard deviation were used to summarise responses to continuous variables as appropriate. Categorical variables were described using frequency tables. Associations between risk factors and low back pain were tested using Pearson’s chi-square test and t-tests as appropriate. A total of 68 questionnaires were completed for analysis (67% participation rate); the period prevalence of LBP was 63.2% (35.0%:beginning of the season, 32.4%:mid-season, 22.1%: end of season). Incidence was 38.2%. The most common location for LBP was the middle low back region (39.5%), and the most common duration of pain was a few hours (32.6%). Most participants (79.1%) did not classify their pain as a disability, and only 44.2% of participants received medical treatment for their LBP. An interesting finding was the association between hydration and LBP (p = 0.050), i.e., those individuals who did not hydrate frequently during matches and training were significantly more likely to experience LBP. The results of this study, although limited to a select group of adolescents, showed a higher prevalence of LBP than that of previous studies. More importantly, even though most participants did not experience LBP classified as a disability, LBP still had a large impact on participants, as nearly half of the participants consulted with a medical professional for treatment. Need for the application of further strategies in the prevention and management of LBP in field hockey, such as adequate warm-up and cool-down, stretching exercises, rest between sessions, etc., are recommended as simple strategies to reduce LBP prevalence.

Keywords: adolescents, field hockey players, incidence, low back pain, prevalence, risk factors

Procedia PDF Downloads 57

Authors: Zi-Yan Liao, Shan-Yu Zhang, Ya-Xian Lin, Ya-Lun Lee, Shih-Chuan Huang, Hong-Ru Lin

Abstract:

Traditional wound dressings, such as gauze, bandages, etc., are easy to adhere to the tissue fluid exuded from the wound, causing secondary damage to the wound during removal. This study takes this as the idea to develop a hydrogel dressing, to explore that the dressing will not cause secondary damage to the wound when it is torn off, and at the same time, create an environment conducive to wound healing. First, the temperature-sensitive material N-isopropylacrylamide (NIPAAm) was used as the substrate. Due to its low mechanical properties, the hydrogel would break due to pulling during human activities. Polyvinyl alcohol (PVA) interpenetrates into it to enhance the mechanical properties, and a semi-interpenetration (semi-IPN) composed of poly(N-isopropylacrylamide) (PNIPAAm) and polyvinyl alcohol (PVA) was prepared by free radical polymerization. PNIPAAm was cross-linked with N,N'-methylenebisacrylamide (NMBA) in an ice bath in the presence of linear PVA, and tetramethylhexamethylenediamine (TEMED) was added as a promoter to speed up the gel formation. The polymerization stage was carried out at 16°C for 17 hours and washed with distilled water for three days after gel formation, and the water was changed several times in the middle to complete the preparation of semi-IPN hydrogel. Finally, various tests were used to analyze the effects of different ratios of PNIPAAm and PVA on semi-IPN hydrogels. In the swelling test, it was found that the maximum swelling ratio can reach about 50% under the environment of 21°C, and the higher the ratio of PVA, the more water can be absorbed. The saturated moisture content test results show that when more PVA is added, the higher saturated water content. The water vapor transmission rate test results show that the value of the semi-IPN hydrogel is about 57 g/m²/24hr, which is not much related to the proportion of PVA. It is found in the LCST test compared with the PNIPAAm hydrogel; the semi-IPN hydrogel possesses the same critical solution temperature (30-35°C). The semi-IPN hydrogel prepared in this study has a good effect on temperature response and has the characteristics of thermal sensitivity. It is expected that after improvement, it can be used in the treatment of surface wounds, replacing the traditional dressing shortcoming.

Keywords: hydrogel, N-isopropylacrylamide, polyvinyl alcohol, hydrogel wound dressing, semi-interpenetrating polymer network

Procedia PDF Downloads 80

Authors: Pronab Ganguly, Ahmed A. Moustafa

Abstract:

Holistic management of schizophrenia involves mainstream pharmacological intervention, complimentary medicine intervention, therapeutic intervention and other psychosocial factors such as accommodation, education, job training, employment, relationship, friendship, exercise, overall well-being, smoking, substance abuse, suicide prevention, stigmatisation, recreation, entertainment, violent behaviour, arrangement of public trusteeship and guardianship, day-day-living skill, integration with community, and management of overweight due to medications and other health complications related to medications amongst others. Our review shows that there is no integrated survey by combining all these factors. An international web-based survey was conducted to evaluate the significance of all these factors and present them in a unified manner. It is believed this investigation will contribute positively towards holistic management of schizophrenia. There will be two surveys. In the pharmacological intervention survey, five popular drugs for schizophrenia will be chosen and their efficacy as well as harmful side effects will be evaluated on a scale of 0 -10. This survey will be done by psychiatrists. In the second survey, each element of therapeutic intervention and psychosocial factors will be evaluated according to their significance on a scale of 0 - 10. This survey will be done by care givers, psychologists, case managers and case workers. For the first survey, professional bodies of psychiatrists in English speaking countries will be contacted to request them to ask their members to participate in the survey. For the second survey, professional bodies of clinical psychologist and care givers in English speaking countries will be contacted to request them to ask their members to participate in the survey. Additionally, for both the surveys, relevant professionals will be contacted through personal contact networks. For both the surveys, mean, mode, median, standard deviation and net promoter score will be calculated for each factor and then presented in a statistically significant manner. Subsequently each factor will be ranked according to their statistical significance. Additionally, country specific variation will be highlighted to identify the variation pattern. The results of these surveys will identify the relative significance of each type of pharmacological intervention, each type of therapeutic intervention and each type of psychosocial factor. The determination of this relative importance will definitely contribute to the improvement in quality of life for individuals with schizophrenia.

Keywords: schizophrenia, holistic management, antipsychotics, quality of life

Procedia PDF Downloads 149

Authors: Mary Carmen Peloche Barrera

Abstract:

Since its early years as an independent nation, the United States has been one of the main promoters regarding the recognition, legislation, and protection of human rights. In the matter of freedom, the founding father Thomas Jefferson envisioned the role of the U.S. as a defender of freedom and equality throughout the world. This founding ideal shaped America’s domestic and foreign policy in the 19th and the 20th century and became an aspiration of the ideals of the country to expand its values and institutions. The history of the emergence of human rights cannot be studied without making reference to leaders such as Woodrow Wilson, Franklin, and Eleanor Roosevelt, as well as Martin Luther King. Throughout its history, this country has proclaimed that the protection of the freedoms of men, both inside and outside its borders, is practically the reason for its existence. Although the United States was one of the first countries to recognize the existence of inalienable rights for individuals, as well as the main promoter of the Universal Declaration of Human Rights of 1948, the country has gone through critical moments that had led to questioning its commitment to the issue. Racial segregation, international military interventions, national security strategy, as well as national legislation on immigration, are some of the most controversial issues related to decisions and actions driven by the United States, which at the same time mismatched with its role as an advocate of human rights, both in the Americas and in the rest of the world. The aim of this paper is to study the swinging of the efforts and commitments of the United States towards human rights. The paper will analyze the history and evolution of human rights in the United States, to study the greatest challenges for the country in this matter. The paper will focus on both the domestic policy (related to demographic issues) and foreign policy (about its role in a post-war world). Currently, more countries are joining the multilateral efforts for the promotion and protection of human rights. At the same time, the United States is one of the least committed countries in this respect, having ratified only 5 of the 18 treaties emanating from the United Nations. The last ratification was carried out in 2002 and, since then, the country has been losing ground, in an increasingly vertiginous way, in its credibility and, even worse, in its role as leader of 'the free world'. With or without the United States, the protection of human rights should remain the main goal of the international community.

Keywords: United States, human rights, foreign policy, domestic policy

Procedia PDF Downloads 117

Authors: Y. Landkocz, C. Lepers, P.J. Martin, B. Fougère, F. Roy Saint-Georges. A. Verdin, F. Cazier, F. Ledoux, D. Courcot, F. Sichel, P. Gosset, P. Shirali, S. Billet

Abstract:

In 2013, the International Agency for Research on Cancer (IARC) classified air pollution and fine particles as carcinogenic to humans. Causal relationships exist between elevated ambient levels of airborne particles and increase of mortality and morbidity including pulmonary diseases, like lung cancer. However, due to a double complexity of both physicochemical Particulate Matter (PM) properties and tumor mechanistic processes, mechanisms of action remain not fully elucidated. Furthermore, because of several common properties between air pollution PM and tobacco smoke, like the same route of exposure and chemical composition, potential mechanisms of synergy could exist. Therefore, smoking could be an aggravating factor of the particles toxicity. In order to identify some mechanisms of action of particles according to their size, two samples of PM were collected: PM0.03 2.5 and PM0.33 2.5 in the urban-industrial area of Dunkerque. The overall cytotoxicity of the fine particles was determined on human bronchial cells (BEAS-2B). Toxicological study focused then on the metabolic activation of the organic compounds coated onto PM and some genetic and epigenetic changes induced on a co-culture model of BEAS-2B and alveolar macrophages isolated from bronchoalveolar lavages performed in smokers and non-smokers. The results showed (i) the contribution of the ultrafine fraction of atmospheric particles to genotoxic (eg. DNA double-strand breaks) and epigenetic mechanisms (eg. promoter methylation) involved in tumor processes, and (ii) the influence of smoking on the cellular response. Three main conclusions can be discussed. First, our results showed the ability of the particles to induce deleterious effects potentially involved in the stages of initiation and promotion of carcinogenesis. The second conclusion is that smoking affects the nature of the induced genotoxic effects. Finally, the in vitro developed cell model, using bronchial epithelial cells and alveolar macrophages can take into account quite realistically, some of the existing cell interactions existing in the lung.

Keywords: air pollution, fine and ultrafine particles, genotoxic and epigenetic alterations, smoking

Procedia PDF Downloads 347

Authors: Nigatu Tadesse, Ying Liu, Juan Li, Hong Ming Liu

Abstract:

Gastric carcinogenesis is a lengthy process of histopathological transition from normal to atrophic gastritis (AG) to intestinal metaplasia (GIM), dysplasia toward gastric cancer (GC). The stage of GIM identified as precancerous lesions with resistance to H-pylori eradication and recurrence after endoscopic surgical resection therapies. GIM divided in to two morphologically distinct phenotypes such as complete GIM bearing intestinal type morphology whereas the incomplete type has colonic type morphology. The incomplete type GIM considered to be the greatest risk factor for the development of GC. Studies indicated the expression of the caudal type homeobox 2 (CDX2) gene is responsible for the development of complete GIM but its progressive downregulation from incomplete metaplasia toward advanced GC identified as the risk for IM progression and neoplastic transformation. The downregulation of CDX2 gene have promoted cell growth and proliferation in gastric and colon cancers and ascribed in chemo-treatment inefficacies. CDX2 downregulated through promoter region hypermethylation in which the methylation frequency positively correlated with the dietary history of the patients, suggesting the role of diet as methyl carbon donor sources such as methionine. However, the metabolism of exogenous methionine is yet unclear. Targeting exogenous methionine metabolism has become a promising approach to limits tumor cell growth, proliferation and progression and increase treatment outcome. This review article discusses molecular alterations that could shed light on the potential of exogenous methionine metabolisms, such as gut microbiota alteration as sources of methionine to host cells, metabolic pathway signaling via PI3K/AKt/mTORC1-c-MYC to rewire exogenous methionine and signature of increased gene methylation index, cell growth and proliferation in GIM, with insights to new treatment avenue via targeting methionine metabolism, and the need for future integrated studies on molecular alterations and metabolomics to uncover altered methionine metabolism and characterization of CDX2 methylation in gastric intestinal metaplasia for potential therapeutic exploitation.

Keywords: altered methionine metabolism, Intestinal metaplesia, CDX2 gene, gastric cancer

Procedia PDF Downloads 86

Authors: Weiliang Gong, Lissa Gomes, Lucile Raymond, Hui Xu, Werner Lutze, Ian L. Pegg

Abstract:

Alkali-activated materials, particularly geopolymers, have attracted much interest in academia. Commercial applications are on the rise, as well. Geopolymers are produced typically by a reaction of one or two aluminosilicates with an alkaline solution at room temperature. Fly ash is an important aluminosilicate source. However, using low-Ca fly ash, the byproduct of burning hard or black coal reacts and sets slowly at room temperature. The development of mechanical durability, e.g., compressive strength, is slow as well. The use of fly ashes with relatively high contents ( > 6%) of unburned carbon, i.e., high loss on ignition (LOI), is particularly disadvantageous as well. This paper will show to what extent these impediments can be mitigated by mixing the fly ash with one or two more aluminosilicate sources. The fly ash used here is generated at the Orlando power plant (Florida, USA). It is low in Ca ( < 1.5% CaO) and has a high LOI of > 6%. The additional aluminosilicate sources are metakaolin and blast furnace slag. Binary fly ash-metakaolin and ternary fly ash-metakaolin-slag geopolymers were prepared. Properties of geopolymer pastes before and after setting have been measured. Fresh mixtures of aluminosilicates with an alkaline solution were studied by Vicat needle penetration, rheology, and isothermal calorimetry up to initial setting and beyond. The hardened geopolymers were investigated by SEM/EDS and the compressive strength was measured. Initial setting (fluid to solid transition) was indicated by a rapid increase in yield stress and plastic viscosity. The rheological times of setting were always smaller than the Vicat times of setting. Both times of setting decreased with increasing replacement of fly ash with blast furnace slag in a ternary fly ash-metakaolin-slag geopolymer system. As expected, setting with only Orlando fly ash was the slowest. Replacing 20% fly ash with metakaolin shortened the set time. Replacing increasing fractions of fly ash in the binary system by blast furnace slag (up to 30%) shortened the time of setting even further. The 28-day compressive strength increased drastically from < 20 MPa to 90 MPa. The most interesting finding relates to the calorimetric measurements. The use of two or three aluminosilicates generated significantly more heat (20 to 65%) than the calculated from the weighted sum of the individual aluminosilicates. This synergetic heat contributes or may be responsible for most of the increase of compressive strength of our binary and ternary geopolymers. The synergetic heat effect may be also related to increased incorporation of calcium in sodium aluminosilicate hydrate to form a hybrid (N,C)A-S-H) gel. The time of setting will be correlated with heat release and maximum heat flow.

Keywords: alkali-activated materials, binary and ternary geopolymers, blends of fly ash, metakaolin and blast furnace slag, rheology, synergetic heats

Procedia PDF Downloads 116

Authors: Natan C. G. Silva, Carlos A. C. Girao Neto, Marcele M. S. Vasconcelos, Luciana R. B. Goncalves, Maria Valderez P. Rocha

Abstract:

Galactooligosaccharides (GOS) are sugars with prebiotic function that can be synthesized chemically or enzymatically, and this last one can be promoted by the action of β-galactosidases. In addition to favoring the transgalactosylation reaction to form GOS, these enzymes can also catalyze the hydrolysis of lactose. A highly studied type of GOS is lactulose because it presents therapeutic properties and is a health promoter. Among the different raw materials that can be used to produce lactulose, whey stands out as the main by-product of cheese manufacturing, and its discarded is harmful to the environment due to the residual lactose present. Therefore, its use is a promising alternative to solve this environmental problem. Thus, lactose from whey is hydrolyzed into glucose and galactose by β-galactosidases. However, in order to favor the transgalactosylation reaction, the medium must contain fructose, due this sugar reacts with galactose to produce lactulose. Then, the glucose-isomerase enzyme can be used for this purpose, since it promotes the isomerization of glucose into fructose. In this scenario, the aim of the present work was first to develop β-galactosidase biocatalysts of Kluyveromyces lactis and to apply it in the integrated reactions of hydrolysis, isomerization (with the glucose-isomerase from Streptomyces murinus) and transgalactosylation reaction, using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% glutaraldehyde was evaluated using different enzymatic loads (2, 5, 7, 10, and 12 mg/g). Subsequently, the hydrolysis and transgalactosylation reactions were studied and conducted at 50°C, 120 RPM for 20 minutes. In parallel, the isomerization of glucose into fructose was evaluated under conditions of 70°C, 750 RPM for 90 min. After, the integration of the three processes for the production of lactulose was investigated. Among the evaluated loads, 7 mg/g was chosen because the best activity of the derivative (44.3 U/g) was obtained, being this parameter determinant for the reaction stages. The other parameters of immobilization yield (87.58%) and recovered activity (46.47%) were also satisfactory compared to the other conditions. Regarding the integrated process, 94.96% of lactose was converted, achieving 37.56 g/L and 37.97 g/L of glucose and galactose, respectively. In the isomerization step, conversion of 38.40% of glucose was observed, obtaining a concentration of 12.47 g/L fructose. In the transgalactosylation reaction was produced 13.15 g/L lactulose after 5 min. However, in the integrated process, there was no formation of lactulose, but it was produced other GOS at the same time. The high galactose concentration in the medium probably favored the reaction of synthesis of these other GOS. Therefore, the integrated process proved feasible for possible production of prebiotics. In addition, this process can be economically viable due to the use of an industrial residue as a substrate, but it is necessary a more detailed investigation of the transgalactosilation reaction.

Keywords: beta-galactosidase, glucose-isomerase, galactooligosaccharides, lactulose, whey

Procedia PDF Downloads 140

Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

Abstract:

The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: glycoproteins, genetic engineering, flocculation, over-expression

Procedia PDF Downloads 415

Authors: Amal Saafan, Kareem Al Sofy, Sameh AbdelGhani, Magdy Amin

Abstract:

Background: Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production of β-lactamases is the objective of the study. Methods: One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar, MicroScan and PCR amplification of blaoxa-51like gene. Acinetobacterbaumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem and cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST, and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like, and blaADC-like genes was performed for isolates that showed, at least, two positive results of three for both AmpC and carbapenemases phenotypic screening tests (obvious activity), in addition to GP3 (for comparison). Detection of blaoxa-51like and blaADC-like genes preceded by ISAba1 was also performed. Results: Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2, and GP3, respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively.AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51like and blaADC-like genes, while blaoxa-23like, blaIMP-like andblaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. Only 12 (44.4%) isolates harbored blaoxa-51like and blaADC-like genes preceded by ISAba1. GP3 isolates showed only positive blaoxa-51like and blaADC-like genes. Conclusion: It is not possible to correlate resistance with presence of blaoxa-51like and blaADC-like genes and presence of ISAba1 was immediate as transcriptional promoter. A blaoxa-23like gene played an important role in carbapenem resistance when compared with blaIMP-like and blaVIM-like gene.

Keywords: acinetobacter, beta-lactams, resistance, antimicrobial agents

Procedia PDF Downloads 345

Authors: Ram V. Mohan, John Rivas-Murillo, Ahmed Mohamed, Wayne D. Hodo

Abstract:

Cementitious materials, an excellent example of highly complex, heterogeneous material systems, are cement-based systems that include cement paste, mortar, and concrete that are heavily used in civil infrastructure; though commonly used are one of the most complex in terms of the material morphology and structure than most materials, for example, crystalline metals. Processes and features occurring at the nanometer sized morphological structures affect the performance, deformation/failure behavior at larger length scales. In addition, cementitious materials undergo chemical and morphological changes gaining strength during the transient hydration process. Hydration in cement is a very complex process creating complex microstructures and the associated molecular structures that vary with hydration. A fundamental understanding can be gained through multi-scale level modeling for the behavior and properties of cementitious materials starting from the material chemistry level atomistic scale to further explore their role and the manifested effects at larger length and engineering scales. This predictive modeling enables the understanding, and studying the influence of material chemistry level changes and nanomaterial additives on the expected resultant material characteristics and deformation behavior. Atomistic-molecular dynamic level modeling is required to couple material science to engineering mechanics. Starting at the molecular level a comprehensive description of the material’s chemistry is required to understand the fundamental properties that govern behavior occurring across each relevant length scale. Material chemistry level models and molecular dynamics modeling and simulations are employed in our work to describe the molecular-level chemistry features of calcium-silicate-hydrate (CSH), one of the key hydrated constituents of cement paste, their associated deformation and failure. The molecular level atomic structure for CSH can be represented by Jennite mineral structure. Jennite has been widely accepted by researchers and is typically used to represent the molecular structure of the CSH gel formed during the hydration of cement clinkers. This paper will focus on our recent work on the shear and compressive deformation and failure behavior of CSH represented by Jennite mineral structure that has been widely accepted by researchers and is typically used to represent the molecular structure of CSH formed during the hydration of cement clinkers. The deformation and failure behavior under shear and compression loading deformation in traditional hydrated CSH; effect of material chemistry changes on the predicted stress-strain behavior, transition from linear to non-linear behavior and identify the on-set of failure based on material chemistry structures of CSH Jennite and changes in its chemistry structure will be discussed.

Keywords: cementitious materials, deformation, failure, material chemistry modeling

Procedia PDF Downloads 286

Authors: Jung-En Kuan, Whei-Fen Wu

Abstract:

In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

Procedia PDF Downloads 133

Authors: Haris Setiawan, Phongsakorn Chuammitri, Korawan Sringarm, Montira Intanon, Anucha Sathanawongs

Abstract:

Capacitation is a prerequisite to achieving sperm competency to penetrate the oocyte naturally occurring in vivo throughout the female reproductive tract and entangling secretory fluid and epithelial cells. One of the crucial compounds in the oviductal fluid which promotes capacitation is albumin, secreted in major concentrations. However, the difficulties in the collection and the inconsistency of the oviductal fluid composition throughout the estrous cycle have replaced its function with serum-based albumins such as bovine serum albumin (BSA). BSA has been primarily involved and evidenced for their stabilizing effect to maintain the acrosome intact during the capacitation process, modulate hyperactivation, and elevate the number of sperm bound to zona pellucida. Contrary to its benefits, the use of blood-derived products in the culture system is not sustainable and increases the risk of disease transmissions, such as Creutzfeldt-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE). Moreover, it has been asserted that this substance is an aeroallergen that produces allergies and respiratory problems. In an effort to identify an alternative sustainable and non-toxic albumin source, the present work evaluated sperm reactions to a capacitation medium containing albumin derived from the flesh of the snakehead fish (Channa striata). Before examining the ability of this non-serum albumin to promote capacitation in bovine sperm, the presence of albumin was detected using bromocresol purple (BCP) at the level of 25% from snakehead fish extract. Following the SDS-PAGE and densitometric analysis, two major bands at 40 kDa and 47 kDa consisting of 57% and 16% of total protein loaded were detected as the potential albumin-related bands. Significant differences were observed in all kinematic parameters upon incubation in the capacitation medium. Moreover, consistently higher values were shown for the kinematic parameters related to hyperactivation, such as amplitude lateral head (ALH), velocity curve linear (VCL), and linearity (LIN) when sperm were treated with 3 mg/mL of snakehead fish albumin among other treatments. Likewise, substantial differences of higher acrosome intact presented in sperm upon incubation with various concentrations of snakehead fish albumin for 90 minutes, indicating that this level of snakehead fish albumin can be used to replace the bovine serum albumin. However, further study is highly required to purify the albumin from snakehead fish extract for more reliable findings.

Keywords: capacitation promoter, snakehead fish, non-serum albumin, bovine sperm

Procedia PDF Downloads 112

Authors: Ayodeji Fasuyi

Abstract:

Telfairia occidentalis is a leafy vegetable commonly grown in the tropics for nutritional benefits. The use of enzymes and probiotics is becoming prominent due to the ban on antibiotics as growth promoters in many parts of the world. It is conceived that with enzymes and probiotics additives, fibrous leafy vegetables can be incorporated into poultry feeds as protein source. However, certain antinutrients were also found in the leaves of Telfairia occidentalis. Four broiler starter and finisher diets were formulated for the two phases of the broiler experiments. A mixture of fiber degrading enzymes, Roxazyme G2 (combination of cellulase, glucanase and xylanase) and probiotics (Turbotox), a growth promoter, were used in broiler diets at 1:1. The Roxazyme G2/Turbotox mixtures were used in diets containing four varying levels of Telfairia occidentalis leaf meal (TOLM) at 0, 10, 20 and 30%. Diets 1 were standard broiler diets without TOLM and Roxazyme G2 and Turbotox additives. Diets 2, 3 and 4 had enzymes and probiotics additives. Certain mineral elements such as Ca, P, K, Na, Mg, Fe, Mn, Cu and Zn were found in notable quantities viz. 2.6 g/100 g, 1.2 g/100 g, 6.2 g/100 g, 5.1 g/100 g, 4.7 g/100 g, 5875 ppm, 182 ppm, 136 ppm and 1036 ppm, respectively. Phytin, phytin-P, oxalate, tannin and HCN were also found in ample quantities viz. 189.2 mg/100 g, 120.1 mg/100 g, 80.7 mg/100 g, 43.1 mg/100 g and 61.2 mg/100 g, respectively. The average weight gain was highest at 46.3 g/bird/day for birds on 10% TOLM diet but similar (P > 0.05) to 46.2 g/bird/day for birds on 20% TOLM. The feed conversion ratio (FCR) of 2.27 was the lowest and optimum for birds on 10% TOLM although similar (P > 0.05) to 2.29 obtained for birds on 20% TOLM. FCR of 2.61 was the highest at 2.61 for birds on 30% TOLM diet. The lowest FCR of 2.27 was obtained for birds on 10% TOLM diet although similar (P > 0.05) to 2.29 for birds on 20% TOLM diet. Most carcass characteristics and organ weights were similar (P > 0.05) for the experimental birds on the different diets except for kidney, gizzard and intestinal length. The values for kidney, gizzard and intestinal length were significantly higher (P < 0.05) for birds on the TOLM diets. The nitrogen retention had the highest value of 72.37 ± 0.10% for birds on 10% TOLM diet although similar (P > 0.05) to 71.54 ± 1.89 obtained for birds on the control diet without TOLM and enzymes/probiotics mixture. There was evidence of a better utilization of TOLM as a plant protein source. The carcass characteristics and organ weights all showed evidence of uniform tissue buildup and muscles development particularly in diets containing 10% of TOLM level. There was also better nitrogen utilization in birds on the 10% TOLM diet. Considering the cheap cost of TOLM, it is envisaged that its introduction into poultry feeds as a plant protein source will ultimately reduce the cost of poultry feeds.

Keywords: Telfairia occidentalis leaf meal, enzymes, probiotics, additives

Procedia PDF Downloads 136

Authors: Michal Pravenec, Miroslava Simakova, Jan Silhavy

Abstract:

Brown adipose tissue (BAT) plays an important role in lipid and glucose metabolism in rodents and possibly also in humans. Recently, using systems genetics approach in the BAT from BXH/HXB recombinant inbred strains, derived from the SHR (spontaneously hypertensive rat) and BN (Brown Norway) progenitors, we identified Cd36 (fatty acid translocase) as the hub gene of co-expression module associated with BAT relative weight and function. An important aspect of BAT biology is to better understand the mechanisms regulating the uptake and utilization of fatty acids and glucose. Accordingly, BAT function in the SHR that harbors mutant nonfunctional Cd36 variant (hereafter referred to as SHR-Cd36⁻/⁻) was compared with SHR transgenic line expressing wild type Cd36 under control of a universal promoter (hereafter referred to as SHR-Cd36⁺/⁺). BAT was incubated in media containing insulin and 14C-U-glucose alone or 14C-U-glucose together with palmitate. Incorporation of glucose into BAT lipids was significantly higher in SHR-Cd36⁺/⁺ versus SHR-Cd36⁻/⁻ rats when incubation media contained glucose alone (SHR-Cd36⁻/⁻ 591 ± 75 vs. SHR-Cd36⁺/⁺ 1036 ± 135 nmol/gl./2h; P < 0.005). Adding palmitate into incubation media had no effect in SHR-Cd36⁻/⁻ rats but significantly reduced glucose incorporation into BAT lipids in SHR-Cd36⁺/⁺ (SHR-Cd36⁻/⁻ 543 ± 55 vs. SHR-Cd36⁺/⁺ 766 ± 75 nmol/gl./2h; P < 0.05 denotes significant Cd36 x palmitate interaction determined by two-way ANOVA). This Cd36-dependent reduced glucose uptake in SHR-Cd36⁺/⁺ BAT was likely secondary to increased palmitate incorporation and utilization due to the presence of wild type Cd36 fatty acid translocase in transgenic rats. This possibility is supported by increased incorporation of 14C-U-palmitate into BAT lipids in the presence of both palmitate and glucose in incubation media (palmitate alone: SHR-Cd36⁻/⁻ 870 ± 21 vs. SHR-Cd36⁺/⁺ 899 ± 42; glucose+palmitate: SHR-Cd36⁻/⁻ 899 ± 47 vs. SHR-Cd36⁺/⁺ 1460 ± 111 nmol/palm./2h; P < 0.05 denotes significant Cd36 x glucose interaction determined by two-way ANOVA). It is possible that addition of glucose into the incubation media increased palmitate incorporation into BAT lipids in SHR-Cd36⁺/⁺ rats because of glucose availability for glycerol phosphate production and increased triglyceride synthesis. These changes in glucose and palmitate incorporation into BAT lipids were associated with significant differential expression of Irs1, Irs2, Slc2a4 and Foxo1 genes involved in insulin signaling and glucose metabolism only in SHR-Cd36⁺/⁺ rats which suggests Cd36-dependent effects on insulin action. In conclusion, these results provide compelling evidence that Cd36 plays an important role in BAT insulin signaling and energy substrate utilization.

Keywords: brown adipose tissue, Cd36, energy substrate utilization, insulin signaling, spontaneously hypertensive rat

Procedia PDF Downloads 139

Authors: Yuanyuan Zhang, Yujuan Zheng, Giuseppina Barutello, Sumako Kameishi, Kungchun Chiu, Katharina Hennig, Martial Balland, Federica Cavallo, Lars Holmgren

Abstract:

Angiogenesis describes that new blood vessels migrate from pre-existing ones to form 3D lumenized structure and remodeling. During directional migration toward the gradient of pro-angiogenic factors, the endothelial cells, especially the tip cells need filopodia to sense the environment and exert the pulling force. Of particular interest are the integrin proteins, which play an essential role in focal adhesion in the connection between migrating cells and extracellular matrix (ECM). Understanding how these biomechanical complexes orchestrate intrinsic and extrinsic forces is important for our understanding of the underlying mechanisms driving angiogenesis. We have previously identified Angiomotin (Amot), a member of Amot scaffold protein family, as a promoter for endothelial cell migration in vitro and zebrafish models. Hence, we established inducible endothelial-specific Amot knock-out mice to study normal retinal angiogenesis as well as tumor angiogenesis. We found that the migration ratio of the blood vessel network to the edge was significantly decreased in Amotec- retinas at postnatal day 6 (P6). While almost all the Amot defect tip cells lost migration advantages at P7. In consistence with the dramatic morphology defect of tip cells, there was a non-autonomous defect in astrocytes, as well as the disorganized fibronectin expression pattern correspondingly in migration front. Furthermore, the growth of transplanted LLC tumor was inhibited in Amot knockout mice due to fewer vasculature involved. By using MMTV-PyMT transgenic mouse model, there was a significantly longer period before tumors arised when Amot was specifically knocked out in blood vessels. In vitro evidence showed that Amot binded to beta-actin, Integrin beta 1 (ITGB1), Fibronectin, FAK, Vinculin, major focal adhesion molecules, and ITGB1 and stress fibers were distinctly induced by Amot transfection. Via traction force microscopy, the total energy (force indicater) was found significantly decreased in Amot knockdown cells. Taken together, we propose that Amot is a novel partner of the ITGB1/Fibronectin protein complex at focal adhesion and required for exerting force transition between endothelial cell and extracellular matrix.

Keywords: angiogenesis, angiomotin, endothelial cell migration, focal adhesion, integrin beta 1

Procedia PDF Downloads 237

Authors: Ala’a H. Al-Muhtaseb, Farrukh Jamil, Sandeep K. Saxena

Abstract:

The fast growth rate of energy consumption along with world population expected to demand 50% more energy by 2030 than nowadays. At present, the energy demand is mostly provided by limited fossil fuel sources such as oil, natural gas, and coal that are resulting in dramatic increase in CO2 emissions from combustion of fossil fuels. The growth of the biodiesel industry over the last decade has resulted in a price drop because glycerol is obtained as a by-product during transesterification of vegetable oil or animal fats, which accounts for one tenth of every gallon of biodiesel produced. The production of oxygenates from glycerol gains much importance due to the excellent diesel-blending property of the oxygenates that not only improve the quality of the fuel but also increases the overall yield of the biodiesel in helping to meet the target for energy production from renewable sources for transport in the energy utilization directives. The reaction of bio-glycerol with bio-acetone was carried out in a magnetically stirred two necked round bottom flaskS. Condensation of bio-glycerol with acetone in the presence of various modified forms of beta zeolite has been done for synthesizing solketal (AB-2 modified with nitric acid, AB-3 modified with oxalic acid). Among all modified forms of beta zeolite, AB-2 showed the best performance for maximum glycerol conversion 94.26 % with 94.21 % solketal selectivity and minimum acetal formation 0.05 %. The physiochemical properties of parent beta zeolite and all its modified forms were analyzed by XRD, SEM, TEM, BET, FTIR and TPD. It has been revealed that AB-2 catalysts with high pore volume and surface area gave high glycerol conversion with maximum solketal selectivity. Despite this, the crystallinity of AB-3 was lower than AB-2 which helps to provide the shorter path length for reactants and product but due high pore volume AB-2 was preferred which gave maximum bio-glycerol conversion. Temperature does matter the glycerol conversion and selectivity of solketal, as it increases from 40 ºC to 60 ºC the conversion of glycerol rises from 80.04 % to 94.26 % and selectivity of solketal from 80.0 % to 94.21 % but further increase in temperature to 100 ºC glycerol conversion reduced to 93.06 % and solketal selectivity to 92.08 %. AB-2 was found to be highly stable as up to 4 repeated experimental runs there was less than 10% decrease in its activity. This process offers an attractive route for converting bio-glycerol, the main by-product of biodiesel to solketal with bio-acetone; a value-added green product with potential industrial applications as a valuable green fuel additive or combustion promoter for gasoline/diesel engines.

Keywords: beta-zeolite, bio-glycerol, catalyst, solketal

Procedia PDF Downloads 214

Authors: Elakneswaran Yogarajah, Toyoharu Nawa, Eiji Owaki

Abstract:

Cement-based materials have been using in various reinforced concrete structural components as well as in nuclear waste repositories. The sulfate attack has been an environmental issue for cement-based materials exposed to sulfate bearing groundwater or soils, and it plays an important role in the durability of concrete structures. The reaction between penetrating sulfate ions and cement hydrates can result in swelling, spalling and cracking of cement matrix in concrete. These processes induce a reduction of mechanical properties and a decrease of service life of an affected structure. It has been identified that the precipitation of secondary sulfate bearing phases such as ettringite, gypsum, and thaumasite can cause the damage. Furthermore, crystallization of soluble salts such as sodium sulfate crystals induces degradation due to formation and phase changes. Crystallization of mirabilite (Na₂SO₄:10H₂O) and thenardite (Na₂SO₄) or their phase changes (mirabilite to thenardite or vice versa) due to temperature or sodium sulfate concentration do not involve any chemical interaction with cement hydrates. Over the past couple of decades, an intensive work has been carried out on sulfate attack in cement-based materials. However, there are several uncertainties still exist regarding the mechanism for the damage of concrete in sulfate environments. In this study, modelling work has been conducted to investigate the chemical degradation of cementitious materials in various sulfate environments. Both internal and external sulfate attack are considered for the simulation. In the internal sulfate attack, hydrate assemblage and pore solution chemistry of co-hydrating Portland cement (PC) and slag mixing with sodium sulfate solution are calculated to determine the degradation of the PC and slag-blended cementitious materials. Pitzer interactions coefficients were used to calculate the activity coefficients of solution chemistry at high ionic strength. The deterioration mechanism of co-hydrating cementitious materials with 25% of Na₂SO₄ by weight is the formation of mirabilite crystals and ettringite. Their formation strongly depends on sodium sulfate concentration and temperature. For the external sulfate attack, the deterioration of various types of cementitious materials under external sulfate ingress is simulated through reactive transport model. The reactive transport model is verified with experimental data in terms of phase assemblage of various cementitious materials with spatial distribution for different sulfate solution. Finally, the reactive transport model is used to predict the long-term performance of cementitious materials exposed to 10% of Na₂SO₄ for 1000 years. The dissolution of cement hydrates and secondary formation of sulfate-bearing products mainly ettringite are the dominant degradation mechanisms, but not the sodium sulfate crystallization.

Keywords: thermodynamic calculations, reactive transport, radioactive waste disposal, PHREEQC

Procedia PDF Downloads 163

Authors: Chen Yang, Jun Hu, Ping Li, Lili Xue

Abstract:

Most companies pay careful attention to consumer feedback collection, so it is popular to find the ‘feedback’ button of all kinds of mobile apps. Yet it is much more changeling to analyze these feedback texts and to catch the true feelings of a consumer regarding either a problem or a complimentary of consumers who hands out the feedback. Especially to the Chinese content, it is possible that; in one context the Chinese feedback expresses positive feedback, but in the other context, the same Chinese feedback may be a negative one. For example, in Chinese, the feedback 'operating with loudness' works well with both refrigerator and stereo system. Apparently, this feedback towards a refrigerator shows negative feedback; however, the same feedback is positive towards a stereo system. By introducing Bradley, M. and Lang, P.'s Affective Norms for English Text (ANET) theory and Bucci W.’s Referential Activity (RA) theory, we, usability researchers at Pingan, are able to decipher the feedback and to find the hidden feelings behind the content. We subtract 2 disciplines ‘valence’ and ‘dominance’ out of 3 of ANET and 2 disciplines ‘concreteness’ and ‘specificity’ out of 4 of RA to organize our own rating system with a scale of 1 to 5 points. This rating system enables us to judge the feelings/emotion behind each feedback, and it works well with both single word/phrase and a whole paragraph. The result of the rating reflects the strength of the feeling/emotion of the consumer when he/she is typing the feedback. In our daily work, we first require a consumer to answer the net promoter score (NPS) before writing the feedback, so we can determine the feedback is positive or negative. Secondly, we code the feedback content according to company problematic list, which contains 200 problematic items. In this way, we are able to collect the data that how many feedbacks left by the consumer belong to one typical problem. Thirdly, we rate each feedback based on the rating system mentioned above to illustrate the strength of the feeling/emotion when our consumer writes the feedback. In this way, we actually obtain two kinds of data 1) the portion, which means how many feedbacks are ascribed into one problematic item and 2) the severity, how strong the negative feeling/emotion is when the consumer is writing this feedback. By crossing these two, and introducing the portion into X-axis and severity into Y-axis, we are able to find which typical problem gets the high score in both portion and severity. The higher the score of a problem has, the more urgent a problem is supposed to be solved as it means more people write stronger negative feelings in feedbacks regarding this problem. Moreover, by introducing hidden Markov model to program our rating system, we are able to computerize the scoring system and are able to process thousands of feedback in a short period of time, which is efficient and accurate enough for the industrial purpose.

Keywords: computerized scoring system, feeling/emotion of consumer feedback, referential activity, text mining

Procedia PDF Downloads 176

Authors: Danna Jia, Bin Li

Abstract:

Background: Atopic dermatitis (AD) is a chronic and refractory inflammatory skin disease characterized by relapsing eczematous and pruritic skin lesions. The global prevalence of AD ranges from 1~ 20%, and its incidence rates are increasing. It affects individuals from infancy to adulthood, significantly impacting their daily lives and social activities. Despite its major health burden, the precise mechanisms underlying AD remain unknown. Understanding the genetic differences associated with AD is crucial for advancing diagnosis and targeted treatment development. This study aims to identify candidate genes of AD by using bioinformatics analysis. Methods: We conducted a comprehensive analysis of four pooled transcriptomic datasets (GSE16161, GSE32924, GSE130588, and GSE120721) obtained from the Gene Expression Omnibus (GEO) database. Differential gene expression analysis was performed using the R statistical language. The differentially expressed genes (DEGs) between AD patients and normal individuals were functionally analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Furthermore, a protein-protein interaction (PPI) network was constructed to identify candidate genes. Results: Among the patient-level gene expression datasets, we identified 114 shared DEGs, consisting of 53 upregulated genes and 61 downregulated genes. Functional analysis using GO and KEGG revealed that the DEGs were mainly associated with the negative regulation of transcription from RNA polymerase II promoter, membrane-related functions, protein binding, and the Human papillomavirus infection pathway. Through the PPI network analysis, we identified eight core genes: CD44, STAT1, HMMR, AURKA, MKI67, and SMARCA4. Conclusion: This study elucidates key genes associated with AD, providing potential targets for diagnosis and treatment. The identified genes have the potential to contribute to the understanding and management of AD. The bioinformatics analysis conducted in this study offers new insights and directions for further research on AD. Future studies can focus on validating the functional roles of these genes and exploring their therapeutic potential in AD. While these findings will require further verification as achieved with experiments involving in vivo and in vitro models, these results provided some initial insights into dysfunctional inflammatory and immune responses associated with AD. Such information offers the potential to develop novel therapeutic targets for use in preventing and treating AD.

Keywords: atopic dermatitis, bioinformatics, biomarkers, genes

Procedia PDF Downloads 82

Authors: Zohar Manber, Ran Elkon

Abstract:

Accumulation of somatic mutations (SMs) in the genome is a major driving force of cancer development. Most SMs in the tumor's genome are functionally neutral; however, some cause damage to critical processes and provide the tumor with a selective growth advantage (termed cancer driver mutations). Current research on functional significance of SMs is mainly focused on finding alterations in protein coding sequences. However, the exome comprises only 3% of the human genome, and thus, SMs in the noncoding genome significantly outnumber those that map to protein-coding regions. Although our understanding of noncoding driver SMs is very rudimentary, it is likely that disruption of regulatory elements in the genome is an important, yet largely underexplored mechanism by which somatic mutations contribute to cancer development. The expression of most human genes is controlled by multiple enhancers, and therefore, it is conceivable that regulatory SMs are distributed across different enhancers of the same target gene. Yet, to date, most statistical searches for regulatory SMs have considered each regulatory element individually, which may reduce statistical power. The first challenge in considering the cumulative activity of all the enhancers of a gene as a single unit is to map enhancers to their target promoters. Such mapping defines for each gene its set of regulating enhancers (termed "set of regulatory elements" (SRE)). Considering multiple enhancers of each gene as one unit holds great promise for enhancing the identification of driver regulatory SMs. However, the success of this approach is greatly dependent on the availability of comprehensive and accurate enhancer-promoter (E-P) maps. To date, the discovery of driver regulatory SMs has been hindered by insufficient sample sizes and statistical analyses that often considered each regulatory element separately. In this study, we analyzed more than 2,500 whole-genome sequence (WGS) samples provided by The Cancer Genome Atlas (TCGA) and The International Cancer Genome Consortium (ICGC) in order to identify such driver regulatory SMs. Our analyses took into account the combinatorial aspect of gene regulation by considering all the enhancers that control the same target gene as one unit, based on E-P maps from three genomics resources. The identification of candidate driver noncoding SMs is based on their recurrence. We searched for SREs of genes that are "hotspots" for SMs (that is, they accumulate SMs at a significantly elevated rate). To test the statistical significance of recurrence of SMs within a gene's SRE, we used both global and local background mutation rates. Using this approach, we detected - in seven different cancer types - numerous "hotspots" for SMs. To support the functional significance of these recurrent noncoding SMs, we further examined their association with the expression level of their target gene (using gene expression data provided by the ICGC and TCGA for samples that were also analyzed by WGS).

Keywords: cancer genomics, enhancers, noncoding genome, regulatory elements

Procedia PDF Downloads 104

Authors: M. Zasada, A. Markiewicz, A. Erkiert-Polguj, E. Budzisz

Abstract:

The epidermis is a keratinized stratified squamous epithelium covered by the hydro-lipid barrier. Therefore, active substances should be able to penetrate through this hydro-lipid coating. L-ascorbic acid is one of the vitamins which plays an important role in stimulation fibroblast to produce collagen type I and in hyperpigmentation lightening. Vitamin C is a water-soluble antioxidant, which protects skin from oxidation damage and rejuvenates photoaged skin. No-needle mesotherapy is a non-invasive rejuvenation technique depending on electric pulses, electroporation, and ultrasounds. These physicals factors result in deeper penetration of cosmetics. It is important to increase the penetration of L-ascorbic acid, thereby increasing the spectrum of its activity. The aim of the work was to assess the effectiveness of pure L-ascorbic acid activity in anti-aging therapy using a needle-free and micro-needling mesotherapy. The study was performed on a group of 35 healthy volunteers in accordance with the Declaration of Helsinki of 1964 and agreement of the Ethics Commissions no RNN/281/16/KE 2017. Women were randomized to mesotherapy or control group. Control group applied topically 2,5 ml serum containing 20% L-ascorbic acid with hydrate from strawberries, every 10 days for a period of 9 weeks. No-needle mesotherapy, on the left half of the face and micro-needling on the right with the same serum, was done in mesotherapy group. The pH of serum was 3.5-4, and the serum was prepared directly prior to the facial treatment. The skin parameters were measured at the beginning and before each treatment. The measurement of the forehead skin was done using Cutometer® (measurement of skin elasticity and firmness), Corneometer® (skin hydration measurement), Mexameter® (skin tone measurement). Also, the photographs were taken by Fotomedicus system. Additionally, the volunteers fulfilled the questionnaire. Serum was tested for microbiological purity and stability after the opening of the cosmetic. During the study, all of the volunteers were taken care of a dermatologist. The regular application of the serum has caused improvement of the skin parameters. Respectively, after 4 and 8 weeks improvement in hydration and elasticity has been seen (Corneometer®, Cutometer® results). Moreover, the number of hyper-pigmentated spots has decreased (Mexameter®). After 8 weeks the volunteers has claimed that the tested product has smoothing and moisturizing features. Subjective opinions indicted significant improvement of skin color and elasticity. The product containing the L-ascorbic acid used with intercellular penetration promoters demonstrates higher anti-aging efficiency than control. In vivo studies confirmed the effectiveness of serum and the impact of the active substance on skin firmness and elasticity, the degree of hydration and skin tone. Mesotherapy with pure L-ascorbic acid provides better diffusion of active substances through the skin.

Keywords: anti-aging, l-ascorbic acid, mesotherapy, promoters

Procedia PDF Downloads 265

Authors: Prateek Bhattacharya

Abstract:

The definition of 'control' in India is a rapidly evolving concept, owing to varying rights attached to varying securities. Shares with differential voting rights (DVRs) provide the holder with differential rights as to voting, as compared to ordinary equity shareholders of the company. Such DVRs can amount to both superior voting rights and inferior voting rights, where DVRs with superior voting rights amount to providing the holder with golden shares in the company. While DVRs are not a novel concept in India having been recognized since 2000, they were placed on a back burner by the Securities and Exchange Board of India (SEBI) in 2010 after issuance of DVRs with superior voting rights was restricted. In June 2019, the SEBI rekindled the ebbing fire of DVRs, keeping mind the fast-paced nature of the global economy, the government's faith that India’s ‘new age technology companies’ (i.e., Start-Ups) will lead the charge in achieving its goal of India becoming a $5 trillion dollar economy by 2024, and recognizing that the promoters of such Start-Ups seek to raise capital without losing control over their companies. DVRs with superior voting rights guarantee promoters with up to 74% shareholding in Start-Ups for a period of 5 years, meaning that the holder of such DVRs can exercise sole control and material influence over the company for that period. This manner of control has the potential of causing both pro-competitive and anti-competitive effects in the markets where these companies operate. On the one hand, DVRs will allow Start-Up promoters/founders to retain control of their companies and protect its business interests from foreign elements such as private/public investors – in a scenario where such investors have multiple investments in firms engaged in associated lines of business (whether on a horizontal or vertical level) and would seek to influence these firms to enter into potential anti-competitive arrangements with one another, DVRs will enable the promoters to thwart such scenarios. On the other hand, promoters/founders who themselves have multiple investments in Start-Ups, which are in associated lines of business run the risk of influencing these associated Start-Ups to engage in potentially anti-competitive arrangements in the name of profit maximisation. This paper shall be divided into three parts: Part I shall deal with the concept of ‘control’, as deliberated upon and decided by the SEBI and the Competition Commission of India (CCI) under both company/securities law and competition law; Part II shall review this definition of ‘control’ through the lens of DVRs, and Part III shall discuss the aforementioned potential pro-competitive and anti-competitive effects caused by the DVRs by examining the current Indian Start-Up scenario. The paper shall conclude by providing suggestions for the CCI to incorporate a clearer and more progressive concept of ‘control’.

Keywords: competition law, competitive effects, control, differential voting rights, DVRs, investor shareholding, merger control, start-ups

Procedia PDF Downloads 123

Authors: E. Poupelloz, S. Gauffinet

Abstract:

Ettringite Ca₆Al₂(SO₄)₃(OH)₁₂26H₂O is one of the major hydrates formed during cement hydration. Ettringite forms in Portland cement from the reaction between tricalcium aluminate Ca₃Al₂O₆ and calcium sulfate. Ettringite is also present in calcium sulfoaluminate cement in which it is the major hydrate, formed by the reaction between yeelimite Ca₄(AlO₂)₆SO₄ and calcium sulfate. About the formation of ettringite, numerous results are available in the literature even if some issues are still under discussion. However, almost all published work about ettringite was done on cementitious systems. Yet in cement, hydration reactions are very complex, the result of dissolution-precipitation processes and are submitted to various interactions. Understanding the formation process of a phase alone, here ettringite, is the first step to later understand the much more complex reactions happening in cement. This study is crucial for the comprehension of early cement hydration and physical behavior. Indeed formation of hydrates, in particular, ettringite, will have an influence on the rheological properties of the cement paste and on the need for admixtures. To make progress toward the understanding of existing phenomena, a specific study of nucleation and growth processes of ettringite was conducted. First ettringite nucleation was studied in ionic aqueous solutions, with controlled but different experimental conditions, as different supersaturation degrees (β), different pH or presence of exogenous ions. Through induction time measurements, interfacial ettringite crystals solution energies (γ) were determined. Growth of ettringite in supersaturated solutions was also studied through chain crystallization reactions. Specific BET surface area measurements and Scanning Electron Microscopy observations seemed to prove that growth process is favored over the nucleation process when ettringite crystals are initially present in a solution with a low supersaturation degree. The influence of stirring on ettringite formation was also investigated. Observation was made that intensity and nature of stirring have a high influence on the size of ettringite needles formed. Needle sizes vary from less than 10µm long depending on the stirring to almost 100µm long without any stirring. During all previously mentioned experiments, initially present ions are consumed to form ettringite in such a way that the supersaturation degree with regard to ettringite is decreasing over time. To avoid this phenomenon a device compensating the drop of ion concentrations by adding some more solutions, and therefore always have constant ionic concentrations, was used. This constant β recreates the conditions of the beginning of cement paste hydration, when the dissolution of solid reagents compensates the consumption of ions to form hydrates. This device allowed the determination of the ettringite precipitation rate as a function of the supersaturation degree β. Taking samples at different time during ettringite precipitation and doing BET measurements allowed the determination of the interfacial growth rate of ettringite in m²/s. This work will lead to a better understanding and control of ettringite formation alone and thus during cements hydration. This study will also ultimately define the impact of ettringite formation process on the rheology of cement pastes at early age, which is a crucial parameter from a practical point of view.

Keywords: cement hydration, ettringite, morphology of crystals, nucleation-growth process

Procedia PDF Downloads 128

Authors: Chii-Dong Ho, Jian-Har Chen

Abstract:

The decline in permeate flux across membrane modules is attributed to the increase in temperature polarization resistance in flat-plate Direct Contact Membrane Distillation (DCMD) modules for pure water productivity. Researchers have discovered that this effect can be diminished by embedding turbulence promoters, which augment turbulence intensity at the cost of increased power consumption, thereby improving vapor permeate flux. The device performance of DCMD modules for permeate flux was further enhanced by shrinking the hydraulic diameters of inserted S-ribs carbon-fiber promoters as well as considering the energy consumption increment. The mass-balance formulation, based on the resistance-in-series model by energy conservation in one-dimensional governing equations, was developed theoretically and conducted experimentally on a flat-plate polytetrafluoroethylene/polypropylene (PTFE/PP) membrane module to predict permeate flux and temperature distributions. The ratio of permeate flux enhancement to energy consumption increment, as referred to an assessment on economic viewpoint and technical feasibilities, was calculated to determine the suitable design parameters for DCMD operations with the insertion of S-ribs carbon-fiber turbulence promoters. An economic analysis was also performed, weighing both permeate flux improvement and energy consumption increment on modules with promoter-filled channels by different array configurations and various hydraulic diameters of turbulence promoters. Results showed that the ratio of permeate flux improvement to energy consumption increment in descending hydraulic-diameter modules is higher than in uniform hydraulic-diameter modules. The fabrication details of the DCMD module filaments implementing the S-ribs carbon-fiber filaments and the schematic configuration of the flat-plate DCMD experimental setup with presenting acrylic plates as external walls were demonstrated in the present study. The S-ribs carbon fibers perform as turbulence promoters incorporated into the artificial hot saline feed stream, which was prepared by adding inorganic salts (NaCl) to distilled water. Theoretical predictions and experimental results exhibited a great accomplishment to considerably achieve permeate flux enhancement, such as the new design of the DCMD module with inserting S-ribs carbon-fiber promoters. Additionally, the Nusselt number for the water vapor transferring membrane module with inserted S-ribs carbon-fiber promoters was generalized into a simplified expression to predict the heat transfer coefficient and permeate flux as well.

Keywords: permeate flux, Nusselt number, DCMD module, temperature polarization, hydraulic diameters

Procedia PDF Downloads 8

Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

Abstract:

The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

Procedia PDF Downloads 227

Authors: Norihiro Kato, Yuriko Takayama

Abstract:

Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

Procedia PDF Downloads 267

Authors: J. Ticona Chambi, E. A. De Almeida, C. A. Andrade Raymundo Gaiotto, A. M. Do Espírito Santo, L. Infantes, S. L. Cuffini

Abstract:

The solubility of active pharmaceutical ingredients (APIs) is challenging for the pharmaceutical industry. The new multicomponent crystalline forms as cocrystal and solvates present an opportunity to improve the solubility of APIs. Commonly, the procedure to obtain multicomponent crystalline forms of a drug starts by screening the drug molecule with the different coformers/solvents. However, it is necessary to develop methods to obtain multicomponent forms in an efficient way and with the least possible environmental impact. The Hansen Solubility Parameters (HSPs) is considered a tool to obtain theoretical knowledge of the solubility of the target compound in the chosen solvent. H-Bond Propensity (HBP), Molecular Complementarity (MC), Coordination Values (CV) are tools used for statistical prediction of cocrystals developed by the Cambridge Crystallographic Data Center (CCDC). The HSPs and the CCDC tools are based on inter- and intra-molecular interactions. The curcumin (Cur), target molecule, is commonly used as an anti‐inflammatory. The demethoxycurcumin (Demcur) and bisdemethoxycurcumin (Bisdcur) are natural analogues of Cur from turmeric. Those target molecules have differences in their solubilities. In this way, the work aimed to analyze and compare different tools for multicomponent forms prediction (solvates) of Cur, Demcur and Biscur. The HSP values were calculated for Cur, Demcur, and Biscur using the chemical group contribution methods and the statistical optimization from experimental data. The HSPmol software was used. From the HSPs of the target molecules and fifty solvents (listed in the HSP books), the relative energy difference (RED) was determined. The probability of the target molecules would be interacting with the solvent molecule was determined using the CCDC tools. A dataset of fifty molecules of different organic solvents was ranked for each prediction method and by a consensus ranking of different combinations: HSP, CV, HBP and MC values. Based on the prediction, 15 solvents were selected as Dimethyl Sulfoxide (DMSO), Tetrahydrofuran (THF), Acetonitrile (ACN), 1,4-Dioxane (DOX) and others. In a starting analysis, the slow evaporation technique from 50°C at room temperature and 4°C was used to obtain solvates. The single crystals were collected by using a Bruker D8 Venture diffractometer, detector Photon100. The data processing and crystal structure determination were performed using APEX3 and Olex2-1.5 software. According to the results, the HSPs (theoretical and optimized) and the Hansen solubility sphere for Cur, Demcur and Biscur were obtained. With respect to prediction analyses, a way to evaluate the predicting method was through the ranking and the consensus ranking position of solvates already reported in the literature. It was observed that the combination of HSP-CV obtained the best results when compared to the other methods. Furthermore, as a result of solvent selected, six new solvates, Cur-DOX, Cur-DMSO, Bicur-DOX, Bircur-THF, Demcur-DOX, Demcur-ACN and a new Biscur hydrate, were obtained. Crystal structures were determined for Cur-DOX, Biscur-DOX, Demcur-DOX and Bicur-Water. Moreover, the unit-cell parameter information for Cur-DMSO, Biscur-THF and Demcur-ACN were obtained. The preliminary results showed that the prediction method is showing a promising strategy to evaluate the possibility of forming multicomponent. It is currently working on obtaining multicomponent single crystals.

Keywords: curcumin, HSPs, prediction, solvates, solubility

Procedia PDF Downloads 63

Authors: S. Candamano, A. Iorfida, F. Crea, A. Macario

Abstract:

Recently, basalt fabric reinforced cementitious composites (FRCM) have attracted great attention because they result to be effective in structural strengthening and cost/environment efficient. In this study, authors investigate their mechanical behavior when an inorganic matrix, belonging to the family of alkali-activated binders, is used. In particular, the matrix has been designed to contain high amounts of industrial by-products and waste, such as Ground Granulated Blast Furnace Slag (GGBFS) and Fly Ash. Fresh state properties, such as workability, mechanical properties and shrinkage behavior of the matrix have been measured, while microstructures and reaction products were analyzed by Scanning Electron Microscopy and X-Ray Diffractometry. Reinforcement is made up of a balanced, coated bidirectional fabric made out of basalt fibres and stainless steel micro-wire, with a mesh size of 8x8 mm and an equivalent design thickness equal to 0.064 mm. Mortars mixes have been prepared by maintaining constant the water/(reactive powders) and sand/(reactive powders) ratios at 0.53 and 2.7 respectively. An appropriate experimental campaign based on direct tensile tests on composite specimens and single-lap shear bond test on brickwork substrate has been thus carried out to investigate their mechanical behavior under tension, the stress-transfer mechanism and failure modes. Tensile tests were carried out on composite specimens of nominal dimensions equal to 500 mm x 50 mm x 10 mm, with 6 embedded rovings in the loading direction. Direct shear tests (DST) were carried out on brickwork substrate using an externally bonded basalt-FRCM composite strip 10 mm thick, 50 mm wide and a bonded length of 300 mm. Mortars exhibit, after 28 days of curing, an average compressive strength of 32 MPa and flexural strength of 5.5 MPa. Main hydration product is a poorly crystalline aluminium-modified calcium silicate hydrate (C-A-S-H) gel. The constitutive behavior of the composite has been identified by means of direct tensile tests, with response curves showing a tri-linear behavior. Test results indicate that the behavior is mainly governed by cracks development (II) and widening (III) up to failure. The ultimate tensile strength and strain were respectively σᵤ = 456 MPa and ɛᵤ= 2.20%. The tensile modulus of elasticity in stage III was EIII= 41 GPa. All single-lap shear test specimens failed due to composite debonding. It occurred at the internal fabric-to-matrix interface, and it was the result of a fracture of the matrix between the fibre bundles. For all specimens, transversal cracks were visible on the external surface of the composite and involved only the external matrix layer. This cracking appears when the interfacial shear stresses increase and slippage of the fabric at the internal matrix layer interface occurs. Since the external matrix layer is bonded to the reinforcement fabric, it translates with the slipped fabric. Average peak load around 945 N, peak stress around 308 MPa and global slip around 6 mm were measured. The preliminary test results allow affirming that Alkali-Activated Materials can be considered a potentially valid alternative to traditional mortars in designing FRCM composites.

Keywords: Alkali-activated binders, Basalt-FRCM composites, direct shear tests, structural strengthening

Procedia PDF Downloads 129

Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

Abstract:

Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

Procedia PDF Downloads 397