Search results for: protein resistance

Authors: Suveen Kumar

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Oral cancer (OC) is the sixth most death causing cancer in world which includes tumour of lips, floor of the mouth, tongue, palate, cheeks, sinuses, throat, etc. Conventionally, the techniques used for OC detection are toluidine blue staining, biopsy, liquid-based cytology, visual attachments, etc., however these are limited by their highly invasive nature, low sensitivity, time consumption, sophisticated instrument handling, sample processing and high cost. Therefore, we developed biosensing chips for non-invasive detection of OC via CYFRA-21-1 biomarker. CYFRA-21-1 (molecular weight: 40 kDa) is secreted in saliva of OC patients which is a non-invasive biological fluid with a cut-off value of 3.8 ng mL-1, above which the subjects will be suffering from oral cancer. Therefore, in first work, 3-aminopropyl triethoxy silane (APTES) functionalized zirconia (ZrO2) nanoparticles (APTES/nZrO2) were used to successfully detect CYFRA-21-1 in a linear detection range (LDR) of 2-16 ng mL-1 with sensitivity of 2.2 µA mL ng-1. Successively, APTES/nZrO2-RGO was employed to prevent agglomeration of ZrO2 by providing high surface area reduced graphene oxide (RGO) support and much wider LDR (2-22 ng mL-1) was obtained with remarkable limit of detection (LOD) as 0.12 ng mL-1. Further, APTES/nY2O3/ITO platform was used for oral cancer bioseneor development. The developed biosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) have wider LDR (0.01-50 ng mL-1) with remarkable limit of detection (LOD) as 0.01 ng mL-1. To improve the sensitivity of the biosensing platform, nanocomposite of yattria stabilized nanostructured zirconia-reduced graphene oxide (nYZR) based biosensor has been developed. The developed biosensing chip having ability to detect CYFRA-21-1 biomolecules in the range of 0.01-50 ng mL-1, LOD of 7.2 pg mL-1 with sensitivity of 200 µA mL ng-1. Further, the applicability of the fabricated biosensing chips were also checked through real sample (saliva) analysis of OC patients and the obtained results showed good correlation with the standard protein detection enzyme linked immunosorbent assay (ELISA) technique.

Keywords: non-invasive, oral cancer, nanomaterials, biosensor, biochip

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Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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In this study, in the breeding Japanese quails (coturnix coturnix japonica), it was aimed to study the effects of cellular insulin receptor stimulation on the performance, some blood parameters, and hatchability features. In the study, a total of 84 breeding quails were used, which are in 6 weeks age, and whose 24 are male and 60 female. In the trial, rations which contain 2900 kcal/kg metabolic energy; crude protein of 20%, and water whose pH is calibrated to 7.45 were administered as ad-libitum, to the animals, as metformin source, metformin-HCl was used and as chrome resource, Chromium Picolinate. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of fodder of 20 mg/kg), and chromium picolinate group (basal ration, added fodder of 1500 ppb Cr. When regarded to the results of performance at the end of trial, it is seen that live weight gain, fodder consumption, egg weight, fodder evaluation coefficient, and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features at the end of trial, it was identified that incubation yield and hatchability are not affected by the treatments but in the groups, in which metformin and chromium picolinate are added to ration, that fertility rose at the significant level compared to control group (p < 0,05). According to the results of blood parameters and hormone at the end of the trial, while the level of plasma glucose level was not affected by treatments (p > 0.05), with the addition of metformin and chromium picolinate to ration, plasma, total control, cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p<0,05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0,05).

Keywords: cholesterol, chromium picolinate, hormone, metformin, performance, quail

Procedia PDF Downloads 182

Authors: Areej A. Alzarqaa, Shahira S. Roushdy, Ali A. Alderfasi, Fahad A. AL-Yahya, Ahmed A. Dawaba

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The resistance of mungbean (Vigna radiata L. Wilczeck) and its physiological responses to drought stress was studied in a greenhouse pot experiment. A randomized complete block Design (RCBD) with factorial arrangement having three replications of each treatment was used. Treatments included three water deficit samples (80%, 40% and 20% of field capacity), two mungbean genotypes (Kawmay-1 and VC2010) and two root-knot nematode (Meloidogyne javanica) infection levels (infected and non-infected). Results showed that water deficit stress significantly hampered most of the studied parameters, except for the shoot water content, whereas genotypes showed highly significant differences for stomatal conductance, shoot dry weight and leaf area. Shoot water content was found to be non-significant in relation to chlorophyll b, shoot dry weight and leaf area, whereas highly significant but negatively correlated with chlorophyll a and stomatal conductance. However, all other possible correlations among studied parameters were found to be highly and positively significant. Results also showed that VC 2010 surpassed Kawmay-1 in most of studied characteristics. In the present study, genotypic variation was observed for these parameters and can be used as a basis for selection of the most promising variety under drought conditions.

Keywords: drought stress, Meloidogyne javanica, mungbean, stomatal conductivity, leaf area, root-knot nematode, shoot water content

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Authors: Jovana Petrovic, Biljana Pajin, Ivana Loncarevic, Aleksandar Fistes, Antun Jozinivic, Durdica Ackar, Drago Subaric

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Recovery of food wastes and their conversion to economically viable products will play a vital role for the management strategies in the years to come. Apple pomace may be considered as wastes, but they contain considerable amounts of high value reusable materials. Apple pomace, the by-product of apple juice and cider production, is a good source of fibre, particularly insoluble one. The remaining apple pulp contains 12% dry residue, which is half dietary fibre. Another remarkable aspect is its richness in polyphenols, components with antioxidant activity. Apple pomace could be an interesting alternative source for fibre and polyphenols in extruded corn meals. The extruded corn meals with the addition of finely ground apple pomace were prepared (the ratio of corn meal: apple pomace was 85:15 and 70:30). Characterization of the extrudates in terms of determining the chemical composition and colour was performed. The color of samples was measured by MINOLTA Chroma Meter CR-400 (Minolta Co., Ltd., Osaka, Japan) using D 65 lighting, a 2º standard observer angle and an 8-mm aperture in the measuring head. The following CIELab color coordinates were determined: L* – lightness, a* – redness to greenness and b* – yellowness to blueness. Protein content decreased significantly from 7.91% to 5.19% with increase in pomace from 0% to 30%, while total fibre content increase from 3.39% to 16.62%. The apple pomace addition produced extrudates with a significantly lower L* value and significantly higher a* value. This study has been fully supported by the Provincial Secretariat for High Education and Scientific Research of the Government of Autonomous Province of Vojvodina, Republic of Serbia, project 142-451-2483/2017 and the Ministry of Science and Technological Development of the Republic of Serbia (Project no. 31014).

Keywords: apple pomace, chemical composition, colour, extruded corn snack products, food waste recovery

Procedia PDF Downloads 203

Authors: Sonal Tuljaram Kame

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Growth and development is based on proper diet. A balanced diet contains all the nutrients in required quantum. Although physical growth is completed by young adulthood, the body tissues remain in a dynamic state with catabolism slightly exceeding anabolism, resulting in a net decrease in the number of cells. After the years of adolescence which cause upheavals in the life of the person, the individual struggle to emerge as an adult who know who he is and what his goals are. During this period nutrients are needed for maintaining the health and energy is required for physical functions and physical activities. The nutritional requirement in young adulthood differs from other periods of life. Iron is needed for haemoglobin synthesis and necessitates by the considerable examination of blood volume. Young adult girls need to ensure adequate intake of iron as they loose 0.5 mg/day by way of menstruation. This is complete awareness about nutritional and health on the other side there is widespread ignorance about nutrition and health among young adult girls. The young adult girls who are aware about nutrition and health seem to be very conscious about nutritional intake and health. Figure consciousness and fear of obesity leads to self imposed intake of nutrients. It may result in various health problems. The study was planned to investigate nutrient intake, find relation between nutritional intake, clinical examination score and health status of young adult girls. The present study is based on the data collected from 120 young adult girls studying in four different competitive exams coaching academies in Akola city of Maharashtra. It was found that nutritional intake of these young adult girls was below the recommended level, nutritional knowledge level and nutritional intake are associated attributes, calories, calcium and protein intake is positively correlated with clinical examination and health status. It was concluded that well planned nutritional counseling for the young adult girls can help prevent nutritional deficiency diseases and disorders which may lead to anaemic condition in young adult girls. Girls need to be educated on intake of iron and vitamin B12.

Keywords: nutritional intake, health status, young adult girls, correlation studies

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Authors: M. Gerasimou, S. Mantzoukis, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Microbial infection of the blood is a serious condition where bacteria invade the bloodstream and cause systemic disease. In such cases, blood cultures are performed. Blood cultures are a key diagnostic test for intensive care unit (ICU) patients. Material and method: The BacT/Alert system, which measures the production of carbon dioxide with metabolic organisms, is used. The positive result in the BacT/Alert system is followed by culture in the following selective media: Blood, Mac Conkey No 2, Chocolate, Mueller Hinton, Chapman and Sabaureaud agar. Gram staining method was used to differentiate bacterial species. The microorganisms were identified by biochemical techniques in the automated Microscan (Siemens) system and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer-based test. Results: In 2017 the Laboratory of Microbiology received 3347 blood cultures. Of these, 170 came from the ICU. 116 found positive. Of these S. epidermidis was identified in 42, A. baumannii in 27, K. pneumoniae in 12 (4 of these KPC ‘Klebsiella pneumoniae carbapenemase’), S. hominis in 8, E. faecium in 7, E. faecalis in 5, P. aeruginosa in 3, C. albicans in 3, S. capitis in 2, K. oxytoca in 2, P. mirabilis in 2, E. coli in 1, S. intermidius in 1 and S. lugdunensis in 1. Conclusions: The study of epidemiological data and microbial resistance phenotypes is essential for the choice of therapeutic regimen for the early treatment and limitation of multivalent strains, while it is a crucial factor to solve diagnostic problems.

Keywords: blood culture, bloodstream, infection, intensive care unit

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Authors: Wenqian Zhang, Xuelin Wang, Yujin Hu, Siyang Wang

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The subsurface layer of a component plays a significant role in its service performance. Any surface mechanical process during fabrication can introduce a deformed layer near the surface, which can be related to the microstructure alteration and strain hardening, and affects the mechanical properties and corrosion resistance of the material. However, there exists a great difficulty in determining the subsurface deformation induced by surface machining. In this study, electron backscatter diffraction (EBSD) was used to study the deformed layer of surface milled 316 stainless steel. The microstructure change was displayed by the EBSD maps and characterized by misorientation variation. The results revealed that the surface milling resulted in heavily nonuniform deformations in the subsurface layer and even in individual grains. The direction of the predominant grain deformation was about 30-60 deg to the machined surface. Moreover, a local deformation rate (LDR) was proposed to quantitatively evaluate the local deformation degree. Both of the average and maximum LDRs were utilized to characterize the deformation trend along the depth direction. It was revealed that the LDR had a strong correlation with the development of grain and sub-grain boundaries. In this work, a scan step size of 1.2 μm was chosen for the EBSD measurement. A LDR higher than 18 deg/μm indicated a newly developed grain boundary, while a LDR ranged from 2.4 to 18 deg/μm implied the generation of a sub-grain boundary. And a lower LDR than 2.4 deg/μm could only introduce a slighter deformation and no sub-grain boundary was produced. According to the LDR analysis with the evolution of grain or sub grain boundaries, the deformed layer could be classified into four zones: grain broken layer, seriously deformed layer, slightly deformed layer and non-deformed layer.

Keywords: surface machining, EBSD, subsurface layer, local deformation

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Authors: Olufemi M. Alabi, Foluke A. Aderemi, Adebayo A. Adewumi, Banwo O. Alabi

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General well-being of animals is of paramount interest in some developed countries and of global importance hence the shift onto alternative housing systems for egg-type chickens as replacement for conventional battery cage system. However, there is paucity of information on the effect of this shift on physiological status of the hens to judge their health via the blood profile. Therefore, investigation was carried out on two strains of hen kept in three different housing systems in humid tropics to evaluate changes in their blood parameters. 108, 17-weeks old super black (SBL) hens and 108, 17-weeks old super brown (SBR) hens were randomly allotted to three different intensive systems Partitioned Conventional Cage (PCC), Extended Conventional Cage (ECC) and Deep Litter System (DLS) in a randomized complete block design with 36 hens per housing system, each with three replicates. The experiment lasted 37 weeks during which blood samples were collected at 18th week of age and bi-weekly thereafter for analyses. Parameters measured are packed cell volume (PCV), hemoglobin concentration (Hb), red blood counts (RBC), white blood counts (WBC) and serum metabolites such as total protein (TP), albumin (Alb), globulin (Glb), glucose, cholesterol, urea, bilirubin, serum cortisol while blood indices such as mean corpuscular hemoglobin (MCH), mean cell volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were calculated. The hematological values of the hens were not significantly (p>0.05) affected by the housing system and strain, so also the serum metabolites except for the serum cortisol which was significantly (p<0.05) affected by the housing system only. Hens housed on PCC had higher values (20.05 ng/ml for SBL and 20.55 ng/ml for SBR) followed by hens on ECC (18.15ng/ml for SBL and 18.38ng/ml for SBL) while hens on DLS had the lowest value (16.50ng/ml for SBL and 16.00ng/ml for SBR) thereby confirming indication of stress with conventionally caged birds. Alternative housing systems can also be adopted for egg-type chickens in the humid tropics from welfare point of view with the results of this work confirming stress among caged hens.

Keywords: blood, housing, humid-tropics, layers

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Authors: Auwal Ibrahim Magashi, Lawan Dan Larai Fagwalawa, Muhammad Bello Ibrahim

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A research was conducted to study genetic variability of some quantitative traits in varieties of cowpea (Vigna unguiculata L. [Walp]) under water stressed from Zaria, Nigeria. Seeds of seven varieties of cowpea (Sampea 1, Sampea 2, IAR1074, Sampea 7, Sampea 8, Sampea 10 and Sampea 12) collected from Institute for Agricultural Research (IAR), Samaru, Zaria were screened for water stressed tolerance. The seeds were then sown in poly bags containing sandy-loam arranged in Completely Randomized Design with three replications for quantitative traits evaluation. The nutritional composition of the seeds obtained from the water stress tolerant varieties of cowpea were analyzed. The result obtained revealed highly significant difference (P ≤ 0.01) in the effects of water stress on the number of wilted and dead plants at 40 days after sowing (DAS) and significant (P ≤ 0.05) 34 DAS. However, sampea 10 has the highest mean performance in terms of number of wilted plants at 34 DAS while sampea 2 and IAR 1074 has the lowest mean performance. However, sampea 7 was found to have the highest mean performance for the number of wilted plants at 40 DAS and sampea 2 is lowest. The result for quantitative traits study indicated highly significant difference (P ≤ 0.01) in the plant height, number of days to 50% flowering, number of days to maturity, number of pods per plant, pod length, number of seeds per plant and 100 seed weight; and significant (P ≤ 0.05) at seedling height and number of branches per plant. Similarly, IAR1074 was found to have high performance in terms of most of the quantitative traits under study. However, sampea 8 has the highest mean performance at nutritional level. It was therefore concluded that, all the seven cowpea genotypes were water stress tolerant and produced considerable yield that contained significant nutrients. It was recommended that IAR1074 should be grown for yield while sampea 8 should be grown for protein supplements.

Keywords: cowpea, genetic variability, quantitative traits, water stress

Procedia PDF Downloads 134

Authors: Al-Naqdi Ibtehal Abdulmonem, Hameed Reem Majeed, Al-Hasnawi Haider Kadhim Salih

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Kindergarten buildings are essential for early childhood education, providing a secure environment for children's development. However, they are susceptible to seismic forces, which can endanger occupants during earthquakes. This article emphasizes the importance of conducting thorough seismic analysis and implementing proper structural design to protect the well-being of children, staff, and visitors. By prioritizing structural integrity and considering functional requirements, engineers can mitigate risks associated with seismic events. The use of specialized software like ETABS is crucial for designing earthquake-resistant kindergartens. An analysis using ETABS software compared the structural performance of two single-story kindergartens in Iraq's Ministry of Education, designed with and without seismic considerations. The analysis aimed to assess the impact of seismic design on structural integrity and safety. The kindergarten was designed with seismic considerations, including moment frames. In contrast, the same kindergarten was analyzed without seismic effects, revealing a lack of structural elements to resist lateral forces, rendering it vulnerable to structural failure during an earthquake. Maximum major shear increased over 4 times and over 5 times for bending moment in both kindergartens designed with seismic considerations induced by lateral loads and seismic forces. This component of shear force is vital for designing elements to resist lateral loads and ensure structural stability.

Keywords: seismic analysis, structural design, lateral loads, earthquake resistance, major shear, ETABS

Procedia PDF Downloads 26

Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

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Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

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Authors: Vera Wilden, Benno Hoffmeister, Georgios Balaskas, Lukas Rauber, Burkhard Walter

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Light-weight timber frame elements represent an efficient structural solution for wooden multistory buildings. The wall elements of such buildings – which act as shear diaphragms- provide lateral stiffness and resistance to wind and seismic loads. The tendency towards multi-story structures leads to challenges regarding the prediction of stiffness, strength and ductility of the buildings. Lightweight timber frame elements are built up of several structural parts (sheeting, fasteners, frame, support and anchorages); each of them contributing to the dynamic response of the structure. This contribution describes the experimental and numerical investigation and development of enhanced lightweight timber frame buildings. These developments comprise high-performance timber frame walls with the variable arrangements of sheathing planes and dissipative anchors at the base of the timber buildings, which reduce damages to the timber structure and can be exchanged after significant earthquakes. In order to prove the performance of the developed elements in the context of a real building a full-scale two-story building core was designed and erected in the laboratory and tested experimentally for its seismic performance. The results of the tests and a comparison of the test results to the predicted behavior are presented. Observation during the test also reveals some aspects of the design and details which need to consider in the application of the timber walls in the context of the complete building.

Keywords: dissipative anchoring, full scale test, push-over-test, wood shear walls

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Authors: Renu Singh

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The caste system in Indian society plays an important role in subjugation of women. It creates divides and controls over women’s sexuality in various ways. This paper attempts to look into various modes in which the institution of caste makes some forms of sexuality as socially “acceptable” norms, while deems others as obscene, immoral and against social ethos. Based on a review of existing literature in this area this paper attempts to understand the notion of sexuality in Indian context. It tries to understand how the emergence of norms and values of sexual behaviour has been entwined with the evolution of caste system and the subjugation of many sections of Indian society. It also attempts to trace the internalisation of patriarchal values in Indian society, and the role played by the colonial rulers in creating and maintaining stringent division of space into public and private ones. It is argued here that brahmanical patriarchy, which is a unique phenomenon of the Indian Subcontinent, plays a crucial role in subjugating and controlling women in general and their sexuality in particular. It also creates a divide among women of different castes. Furthermore, the process of colonisation played an important role in shaping the discourse of sexuality in its present form. There were contradictions as well as consensus between the colonial rulers over the questions of regulation of the private domain, as in introducing reform legislation in the nineteenth century informed the debate on sexuality in postcolonial India. The process of emergence of the dichotomous notions of ‘good’ and ‘bad’ sexuality, and the resistance to any ‘deviation’ from the ‘normal’ sexuality is located, not merely in the ‘passive’ evolution of society, but in the actual politics of it.

Keywords: caste, control, sexuality, regulation, brahmanical patriarchy, India

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Authors: Nondumiso Lushozi, Busisani Lembede, Eliton Chivandi

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Consumption of fructose increases the risk of obesity, nonalcoholic fatty liver disease (NAFLD) and metabolic syndrome in children. Zingerone which is found in ginger has antidiabetic and antiobesogenic properties. Therefore, the aim of the study was to investigate the potential of orally administered zingerone to protect growing Sprague-Dawley rats (mimicking growing children) against high-fructose diet-induced metabolic derangements. Forty, 21-day old female Sprague-Dawley rats were randomly allocated and administered the following four treatments for 12 weeks: group I: standard rat chow (SR) + plain water (PW) + plain gelatine cube (PC). group II: SR + 20% (w/v) fructose solution (FS) + PC. group III: SR + FS + 100 mg/kg/day of fenofibrate in gelatine cube. group IV: SR+ FS + 20 mg/kg/day of zingerone in gelatine cube. The rats’ triglyceride, cholesterol, insulin & adiponectin concentration, visceral fat liver lipid content, homoeostasis model assessment of insulin resistance (HOMA-IR) and ability to handle glucose were determined. Oral administration of zingerone significantly increased (P<0.001) visceral fat and liver lipid content (P<0.001), respectively. Results from the study revealed that administration of 20% fructose solution did not induce metabolic dysfunction, however the zingerone treatment increased visceral fat and liver lipid content, all these lipid abnormalities are typical features of the metabolic syndrome, therefore the current study suggests that zingerone has no effect on metabolic dysfunction in adolescent females.

Keywords: antidiabetic, metabolic syndrome, zingerone, antiobesogenic

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Authors: Taposh Roy, Anna Paradowska, Ralph Abrahams, Quan Lai, Michael Law, Peter Mutton, Mehdi Soodi, Wenyi Yan

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In this investigation, a laser cladding process with a powder feeding was used to deposit stainless steel 410L (high strength, excellent resistance to abrasion and corrosion, and great laser compatibility) onto railhead (higher strength, heat treated hypereutectoid rail grade manufactured in accordance with the requirements of European standard EN 13674 Part 1 for R400HT grade), to investigate the development and controllability of process-induced residual stress in the cladding, heat-affected zone (HAZ) and substrate and to analyse their correlation with hardness profile during two different laser cladding directions (across and along the track). Residual stresses were analysed by neutron diffraction at OPAL reactor, ANSTO. Neutron diffraction was carried out on the samples in longitudinal (parallel to the rail), transverse (perpendicular to the rail) and normal (through thickness) directions with high spatial resolution through the thickness. Due to the thick rail and thin cladding, 4 mm thick reference samples were prepared from every specimen by Electric Discharge Machining (EDM). Metallography across the laser claded sample revealed four distinct zones: The clad zone, the dilution zone, HAZ and the substrate. Compressive residual stresses were found in the clad zone and tensile residual stress in the dilution zone and HAZ. Laser cladding in longitudinally cladding induced higher tensile stress in the HAZ, whereas transversely cladding rail showed lower tensile behavior.

Keywords: laser cladding, residual stress, neutron diffraction, HAZ

Procedia PDF Downloads 245

Authors: Allioua Meryem

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Similar to much of the developing world, Algeria is currently undergoing an epidemiological transition. While mal- and under-nutrition and infectious diseases used to be the main causes of poor health, today there is a higher proportion of chronic, non-communicable diseases (NCDs), including cardiovascular disease, diabetes mellitus, cancer, etc. According to estimates for Algeria from the World Health Organization (WHO), NCDs accounted for 63% of all deaths in 2010. The objective of this study was the assessment of eating habits and anthropometric characteristics in a group of youth aged 15 to 19 years in Tlemcen. This study was conducted on a total effective of 806 youth enrolled in a descriptive cross-sectional study; the classification of nutritional status has been established by international standards IOTF, youth were defined as obese if they had a BMI ≥ 95th percentile, and youth with 85th ≤ BMI ≤ 95th percentile were defined as overweight. Wc is classified by the criteria HD, Wc with moderate risk ≥ 90th percentile and Wc with high risk ≥ 95th percentile. The dietary assessment was based on a 24-hour dietary recall assisted by food records. USDA’S nutrient database for Nutrinux® program was used to analyze dietary intake. Nutrients adequacy ratio was calculated by dividing daily individual intake to dietary recommended intake DRI for each nutrient. 9% of the population was overweight, 3% was obese, 7.5% had abdominal obesity, foods eaten in moderation are chips, cookies, chocolate 1-3 times/day and increased consumption of fried foods in the week, almost half of youth consume sugary drinks more than 3 times per week, we observe a decreased intake of energy, protein (P < 0.001, P = 0.003), SFA (P = 0.018), the NAR of phosphorus, iron, magnesium, vitamin B6, vitamin E, folate, niacin, and thiamin reflecting less consumption of fruit, vegetables, milk, and milk products. Youth surveyed have eating habits at risk of developing obesity and chronic disease.

Keywords: food intake, health, anthropometric characteristics, Algeria

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Authors: Gyati Shilakari Asthana, Abhay Asthana

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1 and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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Authors: Miky Lee, K. Kim, D. Lim, D. Cho

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This paper presents the planning, rationale for test specification derivation, sampling requirements, test facilities, and result analysis used to conduct lifetime expectancy endurance tests on power window switches (PWS) considering thermally induced mechanical stress under diurnal cyclic temperatures during normal operation (power cycling). The detail process of analysis and test results on the selected PWS set were discussed in this paper. A statistical approach to ‘life time expectancy’ was given to the measurement standards dealing with PWS lifetime determination through endurance tests. The approach choice, within the framework of the task, was explained. The present task was dedicated to voltage drop measurement to derive lifetime expectancy while others mostly consider contact or surface resistance. The measurements to perform and the main instruments to measure were fully described accordingly. The failure data from tests were analyzed to conclude lifetime expectancy through statistical method using Weibull cumulative distribution function. The first goal of this task is to develop realistic worst case lifetime endurance test specification because existing large number of switch test standards cannot induce degradation mechanism which makes the switches less reliable. 2nd goal is to assess quantitative reliability status of PWS currently manufactured based on test specification newly developed thru this project. The last and most important goal is to satisfy customer’ requirement regarding product reliability.

Keywords: power window switch, endurance test, Weibull function, reliability, degradation mechanism

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Authors: K. B. M. Islam, Syeeda Shiraj-Um-Mahmuda, Afroj Jahan, A. A. Bhuiyan

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In Bangladesh, the use of imported probiotics in poultry is gradually being increased. But surprisingly, no probiotic bacteria have been isolated yet in Bangladesh despite the existence of scavenging native poultry as potential source that is seemingly more resistant to GIT infection as well as other diseases. Therefore, the study was undertaken to isolate, identify and characterize the potential probiotic Lactobacillus and Bifidobacteria strains from Bangladeshi indigenous poultry, and to evaluate their suitability to use in poultry industry. Crop and cecal samples from 61 healthy indigenous birds were used to isolate potential probiotics strains following conventional cultural methods. A total of 216 isolates were identified following physical, biochemical and molecular methods that belonged to the genus Lactobacillus and Bifidobacteria. An auto-aggregation test was performed for 180 and 136 isolated lactobacilli and bifidobacteria strains, respectively. Twelve lactobacilli isolates and 7 bifidobacteria isolates were selected because of their convenient aggregation. In vitro tests including antibacterial activity, resistance to low pH, hemolytic activities etc. were performed for evaluation of probiotic potential of each strain. Under the in vitro conditions and with respects to the probiotic traits, three lactobacilli; LS16, LS45, LS133 and two bifidobacteria, BS21 and BS90 were found to be potential probiotic strains. Thus, they are proposed to be evaluated for their in vivo probiotic properties. If the proposed strains are found suitable as the probiotics to be used in commercial poultry industry, it is expected that the local probiotics would be more beneficial and would save the huge amount of money that Bangladesh spends every year for the importation of such materials from abroad.

Keywords: Bangladeshi poultry, gut microbiota, lactic acid bacteria, scavenging chicken, GIT health

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Authors: Sarah Nasser, Anne Moreau, Alain Hedoux, Romain Jeantet, Guillaume Delaplace

Abstract:

Background: Spray dryed powders containing some caseins are commonly produced in dairy industry. It is widely admitted that the structure of casein evolves during powder storage, inducing a loss of solubility. However few studies evaluate accurately the destabilization mechanisms at molecular and mesoscopic level, in particular for Native Phospho Casein powder (NPC). Consequently, at the state of the art, it is very difficult to assess which secondary structure change or crosslinks initiate insolubility during storage. To address this issue, controlled ageing conditions have been applied to a NPC powder (which was obtained by spray drying a concentrate containing a higher content of casein (90%), whey protein (8%) and lactose (few %)). Evolution of structure and loss of solubility, with the effects of temperature and time of storage were systematically reported. Methods: FTIR spectroscopy, Raman and Circular Dichroism were used to monitor changes of secondary structure in dry powder and in solution after rehydration. Besides, proteomic tools and electrophoresis have been performed after varying storage conditions for evaluating aggregation and post translational modifications, like lactosylation or phosphorylation. Finally, Tof Sims and MEB were used to follow in parallel evolution of structure in surface and skin formation due to storage. Results + conclusion: These results highlight the important role of storage temperature in the stability of NPC. It is shown that this is not lactosylation at the heart of formation of aggregates, as advanced in others publications This is almost the rise of multitude post translational modifications (chemical cross link), added to disulphide bridges (physical cross link) wich contribute to the destabilisation of structure and aggregation of casein. A relative quantification of each kind of cross link, source of aggregates, is proposed. In addition, it has been proved that migration of lipids and formation of skin in surface during the ageing also explains the evolution of structure casein and thus the alterations of functional properties of NPC powder.

Keywords: casein, cross link, powder, storage

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Authors: Vasilis Andriopoulos, Maria D. Gkioni, Elena Koutra, Savvas G. Mastropetros, Fotini N. Lamari, Sofia Hatziantoniou, Michalis Kornaros

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In the present study, the antioxidant activity of aqueous and methanolic extracts of Chlorella minutissima, Dunaliella salina, Isochrysis galbana, Nannochloropsis oculata and Tisohrysis lutea, as well as the proximate composition and fatty acid profile were evaluated, with the aim to select species suitable for co-production of antioxidants and aquaculture feed. Batch cultivation was performed at 25oC in a modified f/2 medium under continuous illumination and aeration with ambient air. Biomass was collected via centrifugation and extracted first with H2O and subsequently with methanol at two growth phases (early and late stationary). Total phenolic content and antioxidant and reducing activity of the extracts were evaluated. The highest phenolic content was found in the methanolic extract of C. minutissima at the early stationary phase (9.04±0.68 mg Gallic Acid Equivalent g-1 dry weight), and the aqueous extract of D. salina at the late stationary phase (8.78±1.49 mg Gallic Acid Equivalent g-1 Dry weight). Antioxidant activity, measured as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and Ferric reducing antioxidant power assay of methanolic extracts were comparable to the literature and correlated to Total phenolic content and Chlorophyll content of the biomass. No such correlation was found in the aqueous extracts. N. oculata and T. lutea were high in protein (39.88±1.72% Dry weight and 43.30±1.33% Dry weight, respectively) and carotenoids (0.64±0.13% and 0.92±0.02%, respectively). Additionally, they presented high eicosapentaenoic acid and docosahexaenoic acid levels (33.74±9.98 mg eicosapentaenoic acid g-1 DW and 31.31±2.92 mg docosahexaenoic acid g-1 dry weight, respectively). N. oculata and T. lutea are promising candidates for the co-production of antioxidants and aquaculture feed, while C. minutissima and D. salina showed promise due to their higher antioxidant content.

Keywords: aquaculture fee, antioxidant activity, fatty acids, microalgae, total phenolic content

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Authors: Umar Saeed

Abstract:

Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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Authors: Anis Eloud, Sayed Chehaibi

Abstract:

Compaction of agricultural soils as a result of the passage of heavy machinery on the fields is a problem that affects many agronomists and farmers since it results in a loss of yield of most crops. To remedy this, and raise the overall future of the food security challenge, we must study and understand the process of soil degradation. The present review is devoted to understanding the effect of repeated passages on agricultural land. The experiments were performed on a plot of the area of the ESIER, characterized by a clay texture in order to quantify the soil compaction caused by the wheels of the tractor during repeated passages on agricultural land. The test tractor CASE type puissance 110 hp and 5470 kg total mass of 3500 kg including the two rear axles and 1970 kg on the front axle. The state of soil compaction has been characterized by measuring its resistance to penetration by means of a penetrometer and direct manual reading, the density and permeability of the soil. Soil moisture was taken jointly. The measurements are made in the initial state before passing the tractor and after each pass varies from 1 to 7 on the track wheel inflated to 1.5 bar for the rear wheel and broke water to the level of valve and 4 bar for the front wheels. The passages are spaced to the average of one week. The results show that the passage of wheels on a farm tilled soil leads to compaction and the latter increases with the number of passages, especially for the upper 15 cm depth horizons. The first passage is characterized by the greatest effect. However, the effect of other passages do not follow a definite law for the complex behavior of granular media and the history of labor and the constraints it suffers from its formation.

Keywords: wheel traffic, tractor, soil compaction, wheel

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Authors: Z. Okay, M. Kalkan Erdoğan, M. Şahin, M. Saçak

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Polyacrylonitrile (PAN) is one of the most common and cheap fiber-forming polymers because of its high strength and high abrasion resistance properties. The result of alkaline hydrolysis of PAN fiber could be formed the products with conjugated sequences of –C=N–, acrylamide, sodium acrylate, and amidine. In this study, PAN fiber was hydrolyzed in a solution of sodium hydroxide, and this hydrolyzed PAN (HPAN) fiber was used to prepare conductive composite fiber by silver particles. The electrically conductive PAN fiber has the usage potential to produce variety of materials such as antistatic materials, life jackets and static charge reducing products. We monitored the change in the weight loss values of the PAN fiber with hydrolysis time. It was observed that a 60 % of weight loss was obtained in the fiber weight after 7h hydrolysis under the investigated conditions, but the fiber lost its fibrous structure. The hydrolysis time of 5h was found to be suitable in terms of preserving its fibrous structure. The change in the conductivity values of the composite with the preparation conditions such as hydrolysis time, silver ion concentration was studied. PAN fibers with different degrees of hydrolysis were treated with aqueous solutions containing different concentrations of silver ions by continuous stirring at 20 oC for 30 min, and the composite having the maximum conductivity of 2 S/cm could be prepared. The antibacterial property of the conductive HPAN fibers participated silver was also investigated. While the hydrolysis of the PAN fiber was characterized with FTIR and SEM techniques, the silver reduction process of the HPAN fiber was investigated with SEM and TGA-DTA techniques. The SEM micrographs showed that the surface of HPAN fiber was rougher and much more corroded than that of the PAN fiber. Composite, Conducting polymer, Fiber, Polyacrylonitrile.

Keywords: composite, conducting polymer, fiber, polyacrylonitrile

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Authors: Jiangang Shen

Abstract:

Olfactory dysfunction is a common symptom companied by anxiety- and depressive-like behaviors in depressive patients. Chronic stress triggers hormone responses and inhibits the proliferation and differentiation of neural stem cells (NSCs) in the hippocampus and subventricular zone (SVZ)-olfactory bulb (OB), contributing to depressive behaviors and olfactory dysfunction. However, the cellular signaling molecules to regulate chronic stress mediated olfactory dysfunction are largely unclear. Adaptor proteins containing the pleckstrin homology domain, phosphotyrosine binding domain, and leucine zipper motif (APPLs) are multifunctional adaptor proteins. Herein, we tested the hypothesis that APPL2 could inhibit hippocampal neurogenesis by affecting glucocorticoid receptor (GR) signaling, subsequently contributing to depressive and anxiety behaviors as well as olfactory dysfunctions. The major discoveries are included: (1) APPL2 Tg mice had enhanced GR phosphorylation under basic conditions but had no different plasma corticosterone (CORT) level and GR phosphorylation under stress stimulation. (2) APPL2 Tg mice had impaired hippocampal neurogenesis and revealed depressive and anxiety behaviors. (3) GR antagonist RU486 reversed the impaired hippocampal neurogenesis in the APPL2 Tg mice. (4) APPL2 Tg mice displayed higher GR activity and less capacity for neurogenesis at the olfactory system with lesser olfactory sensitivity than WT mice. (5) APPL2 negatively regulates olfactory functions by switching fate commitments of NSCs in adult olfactory bulbs via interaction with Notch1 signaling. Furthermore, baicalin, a natural medicinal compound, was found to be a promising agent targeting APPL2/GR signaling and promoting adult neurogenesis in APPL2 Tg mice and chronic corticosterone-induced depression mouse models. Behavioral tests revealed that baicalin had antidepressant and olfactory-improving effects. Taken together, APPL2 is a critical therapeutic target for antidepressant treatment.

Keywords: APPL2, hippocampal neurogenesis, depressive behaviors and olfactory dysfunction, stress

Procedia PDF Downloads 52

Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

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In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and Pep@AuNC, therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the Pep@AuNC conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the Pep@AuNC. To our best knowledge the fabrication of an optical biosensor based on Pep@AuNC for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

Procedia PDF Downloads 144

Authors: Bhavatharani S., Boopathy V., Kavin S., Naveethkumar R.

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Context: Resistance training with blood flow restriction (BFR) has increased in clinical rehabilitation due to the substantial benefits observed in augmenting muscle mass and strength using low loads. However, there is a great variability of training pressures for clinical populations as well as methods to estimate it. The aim of this study was to estimate the percentage of maximal BFR that could result by applying different methodologies based on arbitrary or individual occlusion levels using a cuff width between 9 and 13 cm. Design: A secondary analysis was performed on the combined databases of 2 previous larger studies using BFR training. Methods: To estimate these percentages, the occlusion values needed to reach complete BFR (100% limb occlusion pressure [LOP]) were estimated by Doppler ultrasound. Seventy-five participants (age 24.32 [4.86] y; weight: 78.51 [14.74] kg; height: 1.77 [0.09] m) were enrolled in the laboratory study for measuring LOP in the thigh, arm, or calf. Results: When arbitrary values of restriction are applied, a supra-occlusive LOP between 120% and 190% LOP may result. Furthermore, the application of 130% resting brachial systolic blood pressure creates a similar occlusive stimulus as 100% LOP. Conclusions: Methods using 100 mm Hg and the resting brachial systolic blood pressure could represent the safest application prescriptions as they resulted in applied pressures between 60% and 80% LOP. One hundred thirty percent of the resting brachial systolic blood pressure could be used to indirectly estimate 100% LOP at cuff widths between 9 and 13 cm. Finally, methodologies that use standard values of 200 and, 300 mm Hg far exceed LOP and may carry additional risk during BFR exercise.

Keywords: lower limb rehabilitation, ESP32, pneumatics for medical, programmed rehabilitation

Procedia PDF Downloads 57

Authors: Raya Harkouss, Bilal Hamad

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Self-Consolidating Concrete (SCC) is considered as a relatively new technology created as an effective solution to problems associated with low quality consolidation. A SCC mix is defined as successful if it flows freely and cohesively without the intervention of mechanical compaction. The construction industry is showing high tendency to use SCC in many contemporary projects to benefit from the various advantages offered by this technology. At this point, a main question is raised regarding the effect of enhanced fluidity of SCC on the structural behavior of high strength self-consolidating reinforced concrete. A three phase research program was conducted at the American University of Beirut (AUB) to address this concern. The first two phases consisted of comparative studies conducted on concrete and mortar mixes prepared with second generation Sulphonated Naphtalene-based superplasticizer (SNF) or third generation Polycarboxylate Ethers-based superplasticizer (PCE). The third phase of the research program investigates and compares the structural performance of high strength reinforced concrete beam specimens prepared with two different generations of superplasticizers that formed the unique variable between the concrete mixes. The beams were designed to test and exhibit flexure, shear, or bond splitting failure. The outcomes of the experimental work revealed comparable resistance of beam specimens cast using self-compacting concrete and conventional vibrated concrete. The dissimilarities in the experimental values between the SCC and the control VC beams were minimal, leading to a conclusion, that the high consistency of SCC has little effect on the flexural, shear and bond strengths of concrete members.

Keywords: self-consolidating concrete (SCC), high-strength concrete, concrete admixtures, mechanical properties of hardened SCC, structural behavior of reinforced concrete beams

Procedia PDF Downloads 226

Authors: Valentina Narvaez Gaitan, Paula K. Rodriguez Ramirez, Derian D. Espinosa

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Corrective orthosis has great importance in orthopedic treatments providing assistance in improving mobility and stability in order to improve the quality of life for a different patient. The corrective orthosis studied in this article can correct deformities, reduce pain, and improve the ability to perform daily activities. This work describes the design and evaluation of a corrective orthosis for knee hyperextension. This orthosis is capable of generating a progressive and variable alignment of the joint, limiting the range of motion according to medical criteria. The main objective was to design a corrective knee orthosis capable of correcting knee hyperextension progressively to return to its natural angle with greater economic affordability and adjustable size. The limiting mechanism is based on a goniometer to determine the desired angles. The orthosis was made of acrylic to reduce costs and maintenance; neoprene is also used to make comfortable contact; additionally, Velcro was used in order to adjust the orthosis for various sizes. Simulations of static and fatigue analysis of the mechanism were performed to verify its resistance and durability under normal conditions. A biomechanical gait study of gait was carried out on 10 healthy subjects without the orthosis and limiting their knee extension capacity in a normal gait cycle with the orthosis to observe the efficiency of the proposed system. In the results obtained, the knee angle curves show that the maximum extension angle was the established angle by the orthosis. Showing the efficiency of the proposed design for different leg sizes.

Keywords: biomechanical study, corrective orthosis, efficiency, goniometer, knee hyperextension.

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Authors: Ahmad Ali Shahid, M Shakil Shaukat

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Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.

Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR

Procedia PDF Downloads 364