Search results for: head and neck squamous cell carcinoma

Authors: Gourav Jain, Sameer Dixit, Surjeet Kumar Arya, Praveen C. Verma

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Plant cell wall plays a major role in defence mechanism against biotic and abiotic stress as it constitutes the physical barrier between the microenvironment and internal component of the cell. It is a complex structure composed of mostly carbohydrates among which cellulose and hemicelluloses are most abundant that is embedded in a matrix of pectins and proteins. Multiple enzymes have been reported which plays a vital role in cell wall modification, Pectin Methylesterase (PME) is one of them which catalyses the demethylesterification of homogalacturonans component of pectin which releases acidic pectin and methanol. As emitted methanol is toxic to the insect pest, we use PME gene for the better methanol production. In the current study we showed overexpression of PME gene isolated from Withania somnifera under the insect inducible promoter causes enhancement of methanol production at the time of insect feeds to plants, and that provides better insect resistance property. We found that the 85-90% mortality causes by transgenic tobacco in both chewing (Spodoptera litura larvae and Helicoverpa armigera) and sap-sucking (Aphid, mealybug, and whitefly) pest. The methanol content and emission level were also enhanced by 10-15 folds at different inducible time point interval (15min, 30min, 45min, 60min) which would be analysed by Purpald/Alcohol Oxidase method.

Keywords: methanol, Pectin methylesterase, inducible promoters, Purpald/Alcohol oxidase

Procedia PDF Downloads 242

Authors: Andrew White, Abigail Lowe, Kory Watkins, Hamed Akhlaghi, Nicole Winter

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The timing and appropriateness of diagnostic imaging are critical to the evaluation and management of traumatic injuries. Within the subclass of trauma patients, the prevalence of c-spine injury is less than 4%. However, the incidence of delayed diagnosis within this cohort has been documented as up to 20%, with inadequate radiological examination most cited issue. In order to assess those in which c-spine injury cannot be fully excluded based on clinical examination alone and, therefore, should undergo diagnostic imaging, a set of criteria is used to provide clinical guidance. The NEXUS (National Emergency X-Radiography Utilisation Study) criteria is a validated clinical decision-making tool used to facilitate selective c-spine radiography. The criteria allow clinicians to determine whether cervical spine imaging can be safely avoided in appropriate patients. The NEXUS criteria are widely used within the Emergency Department setting given their ease of use and relatively straightforward application and are used in the Victorian State Trauma System’s guidelines. This audit utilized retrospective data collection to examine the concordance of c-spine imaging in trauma patients to that of the NEXUS criteria and assess compliance with state guidance on diagnostic imaging in trauma. Of the 183 patients that presented with trauma to the head, neck, or face (244 excluded due to incorrect triage), 98 did not undergo imaging of the c-spine. Out of those 98, 44% fulfilled at least one of the NEXUS criteria, meaning the c-spine could not be clinically cleared as per the current guidelines. The criterion most met was intoxication, comprising 42% (18 of 43), with midline spinal tenderness (or absence of documentation of this) the second most common with 23% (10 of 43). Intoxication being the most met criteria is significant but not unexpected given the cohort of patients seen at St Vincent’s and within many emergency departments in general. Given these patients will always meet NEXUS criteria, an element of clinical judgment is likely needed, or concurrent use of the Canadian C-Spine Rules to exclude the need for imaging. Midline tenderness as a met criterion was often in the context of poor or absent documentation relating to this, emphasizing the importance of clear and accurate assessments. The distracting injury was identified in 7 out of the 43 patients; however, only one of these patients exhibited a thoracic injury (T11 compression fracture), with the remainder comprising injuries to the extremities – some studies suggest that C-spine imaging may not be required in the evaluable blunt trauma patient despite distracting injuries in any body regions that do not involve the upper chest. This emphasises the need for standardised definitions for distracting injury, at least at a departmental/regional level. The data highlights the currently poor application of the NEXUS guidelines, with likely common themes throughout emergency departments, highlighting the need for further education regarding implementation and potential refinement/clarification of criteria. Of note, there appeared to be no significant differences between levels of experience with respect to inappropriately clearing the c-spine clinically with respect to the guidelines.

Keywords: imaging, guidelines, emergency medicine, audit

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Authors: Majid Moshirfar, Kyle Margulies, Yasmyne C. Ronquillo, Phillip Hoopes

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A total of 66 articles were reviewed to compare glaucoma and its associated surgeries' effect on central corneal endothelium cell density (CECD). The paper reports the average reported central CECD loss at 3-, 6-, 12-, 24-, 36-, 48-, and 60-month post-operation for each glaucoma surgery. ALT, MLT, SLT, CS AGV, VC BGI, Hydrus + phaco, XEN gel + phaco, PRESERFLO, Dual iStent, or Trabectome had no significant impact on postoperative CECD compared to either preoperative CECD or control group CECD. The highest CECD loss was found to be EXPRESS-phaco, AC AGV, CS BGI, CS BGI, AC BGI, and AC BGI at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups, respectively. AC AGV, Trab + MMC, Trab, AC BGI, Trab + MMC, Cypass, and Cypass showed the smallest reduction of CECD at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups.

Keywords: glaucoma, corneal endothelium, cell density, surgery outcome

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Authors: Urszula Stachewicz

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Surface properties of materials are the key parameter in many applications, especially in the biomedical field, to control cell-material interactions. In our work, we want to achieve the controllability of surface properties of polymer fibers via a single-step electrospinning process by alternating voltage polarities. Voltage polarity defines the charge accumulated on the surface of the liquid jet and the surface of the fibers. Positive polarity attracts negatively charged groups to fibers’ surface, whereas negative polarity moves the negatively charged functional groups away from the surface. This way, we can control the surface chemistry, wettability, and additionally surface potential of electrospun fibers. Within our research, we characterized surface chemistry using X-ray photoelectron microscopy (XPS) and surface potential with Kelvin probe force microscopy (KPFM) on electrospun fibers of commonly used polymers such as PCL, PVDF, and PMMA, often used as biomaterials. We proved the significant effect of fibers' surface potential on cell integration with the scaffolds and further cells development for the regeneration processes based on the osteoblast and fibroblast culture studies. Acknowledgments: The study was conducted within ‘Nanofiber-based sponges for atopic skin treatment’ project, which is carried out within the First TEAM programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund, project no POIR.04.04.00-00- 4571/18-00.

Keywords: cell attachment, fibers, fibroblasts, osteoblast, proliferation, surface potential

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Authors: G. A. Amin, A. D. Al-Talhi

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The bacterium B. subtilis produced surfactin in conventional batch culture as a growth associated product and a growth rate (0.4 h-1). A fed-batch process was developed and the fermentative substrate and other nutrients were fed on hourly basis and according to the growth rate of the bacterium. Conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with fermentation run supplied with 200 g Maldex-15. Up to 35.4 g.l-1 of surfactin and cell biomass of 30.2 g.l-1 were achieved in 12 hrs. Also, markedly substrate yield of 0.269 g/g and volumetric reactor productivity of 2.61 g.1-1.h-1 were obtained confirming the establishment of a cost effective commercial surfactin production.

Keywords: Bacillus subtilis, biosurfactant, exponentially fed-batch fermentation, surfactin

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Authors: S. Vasailor, C. Rattanakawin

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Electro-winning of copper metal from dilute sulfate solution (13.7 g/L) was performed in a lab electrolytic cell with stainless-steel cathode and lead-alloy anode. The effects of various parameters including cell voltage, electro-winning temperature and time were studied in order to acquire an appropriate current efficiency of copper deposition. The highest efficiency is about 95% obtaining from electro-winning condition of 3V, 55°C and 3,600 s correspondingly. The cathode copper with 95.5% Cu analyzed using atomic absorption spectrometry can be obtained from this single-winning condition. In order to increase the copper grade, solvent extraction should be used to increase the sulfate concentration, say 50 g/L, prior to winning the cathode copper effectively.

Keywords: copper metal, current efficiency, dilute sulfate solution, electro-winning

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Authors: Mahdi Eskandarlou, Mehrdad Taghipour

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Background: Partial-thickness skin graft is the cornerstone for scalp defect repair. Routine donor sites include abdomen, thighs, and buttocks. Given the potential side effects following harvesting from these sites and the potential advantages of harvesting from scalp (broad surface, rapid healing, and better cosmetics results), this study is trying to compare the outcomes of graft harvesting from scalp and lower limb. Methods: This clinical trial is conducted among a sample number of 40 partial thickness graft candidates (20 case and 20 control group) with scalp defect presenting to plastic surgery clinic at Besat Hospital during the time period between 2018 and 2019. Sampling was done by simple randomization using random digit table. Data gathering was performed using a designated checklist. The donor site in case group and control group was scalp and lower limb, respectively. The resultant data were analyzed using chi-squared and t-test and SPPS version 21 (SPSS Statistics for Windows, Version 21.0. Armonk, NY: IBM Corp). Results: Of the total 40 patients participating in this study, 28 patients (70%) were male, and 12 (30%) were female with and mean age of 63.62 ± 09.73 years. Hypertension and diabetes mellitus were the most common comorbidities among patients with basal cell carcinoma (BCC) and trauma being the most common etiology for the defects. There was a statistically meaningful relationship between two groups regarding the etiology of defect (P=0.02). The most common anatomic location of defect for case and control groups was temporal and parietal, respectively. Most of the defects were deep to galea zone. The mean diameter of defect was 24.28 ± 45.37 mm for all of the patients. The difference between diameter of defect in both groups was statistically meaningful, while no such difference between graft diameter was seen. The graft 'Take' was completely successful in both groups according to evaluations. The level of postoperative pain was lower in the case group compared to the control according to VAS scale, and the satisfaction was higher in them per Likert scale. Conclusion: Scalp can safely be used as donor site for skin graft to be used for scalp defects, which is associated with better results and lower complication rates compared to other donor sites.

Keywords: donor site, leg, partial-thickness graft, scalp

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Authors: Orod Zarrin, Mohesn Ramezan Shirazi, Hassan Moniri

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The use of pile foundations technique is developed to support structures and buildings on soft soil. The most important dynamic load that can affect the pile structure is earthquake vibrations. Pile foundations during earthquake excitation indicate that piles are subject to damage by affecting the superstructure integrity and serviceability. During an earthquake, two types of stresses can damage the pile head, inertial load that is caused by superstructure and deformation which caused by the surrounding soil. Soil deformation and inertial load are associated with the acceleration developed in an earthquake. The acceleration amplitude at the ground surface depends on the magnitude of earthquakes, soil properties and seismic source distance. According to the investigation, the damage is between the liquefiable and non-liquefiable layers and also soft and stiff layers. This damage crushes the pile head by increasing the inertial load which is applied by the superstructure. On the other hand, the cracks on the piles due to the surrounding soil are directly related to the soil profile and causes cracks from small to large. However, the large cracks reason have been listed such as liquefaction, lateral spreading, and inertial load. In the field of designing, elastic response of piles is always a challenge for designer in liquefaction soil, by allowing deflection at top of piles. Moreover, absence of plastic hinges in piles should be insured, because the damage in the piles is not observed directly. In this study, the performance and behavior of pile foundations during liquefaction and lateral spreading are investigated. In addition, emphasize on the soil behavior in the liquefiable and non-liquefiable layers by different aspect of piles damage such as ranking, location and degree of damage are going to discuss.

Keywords: pile, earthquake, liquefaction, non-liquefiable, damage

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Authors: Ateeqa Aslam, Hans J. Nauwynck

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Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that has been studied in the past as a cause of vomiting and wasting disease (VWD) in young piglets (<3 weeks). Nowadays, the virus is still circulating on most farms in Belgium, but there are no descriptions anymore of VWD. Therefore, we are interested in differences between the old and new strains. We compared the replication kinetics of the old well-studied strain VW572 (1972) and the recent isolate P412 (2020) in a susceptible continuous cell line (RPD cells) and in primary porcine respiratory epithelial cells (PoRECs). The RPD cell line was inoculated with each PHEV strain at an m.o.i. of 1 the supernatant was collected, and the cells were fixed at different time points post-inoculation. The supernatant was titrated (extracellular virus titer), and the infected cells were revealed by immunofluorescence staining and quantitated by fluorescence microscopy. We found that VW572 replicated better in the RPD cell line at earlier time points when compared to P412. Porcine respiratory epithelial cells (PoREC) were isolated, grown at air-liquid interphase in transwells and inoculated with both strains of PHEV at a virus titer of 106.6TCID50 per 200 µl either at the apical side or at the basal side of the cells. At different time points after inoculation, the transwells were fixed and stained for infected cells. VW572 preferentially infected the epithelial cells via the basolateral side of porcine nasal epithelial cells, whereas P412 preferred the apical side. These findings suggest that there has been an evolution of PHEV in its interaction with the respiratory epithelial cells. In the future, more virus strains will be enclosed and the tropism of the strains for different neuronal cell types will be examined for the change in virus neurotropism.

Keywords: porcine hemagglutinating encephalomyelitis virus (PHEV), primary porcine respiratory epithelial cells (PoRECs), virus tropism, vomiting and wasting disease (VWD)

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Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

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Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

Procedia PDF Downloads 339

Authors: I. Székács, Á. Fejes, S. Klátyik, E. Takács, D. Patkó, J. Pomóthy, M. Mörtl, R. Horváth, E. Madarász, B. Darvas, A. Székács

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Environmental and toxicological characteristics of formulated pesticides may substantially differ from those of their active ingredients or other components alone. This phenomenon is demonstrated in the case of the herbicide active ingredient glyphosate. Due to its extensive application, this active ingredient was found in surface and ground water samples collected in Békés County, Hungary, in the concentration range of 0.54–0.98 ng/ml. The occurrence of glyphosate appeared to be somewhat higher at areas under intensive agriculture, industrial activities and public road services, but the compound was detected at areas under organic (ecological) farming or natural grasslands, indicating environmental mobility. Increased toxicity of the formulated herbicide product Roundup, compared to that of glyphosate was observed on the indicator aquatic organism Daphnia magna Straus. Acute LC50 values of Roundup and its formulating adjuvant Polyethoxylated Tallowamine (POEA) exceeded 20 and 3.1 mg/ml, respectively, while that of glyphosate (as isopropyl salt) was found to be substantially lower (690-900 mg/ml) showing good agreement with literature data. Cytotoxicity of Roundup, POEA and glyphosate has been determined on the neuroectodermal cell line, NE-4C measured both by cell viability test and holographic microscopy. Acute toxicity (LC50) of Roundup, POEA and glyphosate on NE-4C cells was found to be 0.013±0.002%, 0.017±0.009% and 6.46±2.25%, respectively (in equivalents of diluted Roundup solution), corresponding to 0.022±0.003 and 53.1±18.5 mg/ml for POEA and glyphosate, respectively, indicating no statistical difference between Roundup and POEA and 2.5 orders of magnitude difference between these and glyphosate. The same order of cellular toxicity seen in average cell area has been indicated under quantitative cell visualization. The results indicate that toxicity of the formulated herbicide is caused by the formulating agent, but in some parameters toxicological synergy occurs between POEA and glyphosate.

Keywords: glyphosate, polyethoxylated tallowamine, Roundup, combined aquatic and cellular toxicity, synergy

Procedia PDF Downloads 317

Authors: R. J. Pontawe, R. C. Martinez, N. T. Asuncion, R. V. Villacorte

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Drying of high moisture paddy remains a major problem in the Philippines, especially during inclement weather condition. To alleviate the problem, mechanical dryers were used like a flat bed and recirculating batch-type dryers. However, drying to 14% (wet basis) final moisture content is long which takes 10-12 hours and tedious which is not the ideal for handling high moisture paddy. Fully-automated pilot-scale fluidized bed drying system with 500 kilograms per hour capacity was evaluated using a high moisture paddy. The developed fluidized bed dryer was evaluated using four drying temperatures and two variations in fluidization time at a constant airflow, static pressure and tempering period. Complete drying of paddy with ≥28% (w.b.) initial MC was attained after 2 passes of fluidized-bed drying at 2 minutes exposure to 70 °C drying temperature and 4.9 m/s superficial air velocity, followed by 60 min ambient air tempering period (30 min without ventilation and 30 min with air ventilation) for a total drying time of 2.07 h. Around 82% from normal mechanical drying time was saved at 70 °C drying temperature. The drying cost was calculated to be P0.63 per kilogram of wet paddy. Specific heat energy consumption was only 2.84 MJ/kg of water removed. The Head Rice Yield recovery of the dried paddy passed the Philippine Agricultural Engineering Standards. Sensory evaluation showed that the color and taste of the samples dried in the fluidized bed dryer were comparable to air dried paddy. The optimum drying parameters of using fluidized bed dryer is 70 oC drying temperature at 2 min fluidization time, 4.9 m/s superficial air velocity, 10.16 cm grain depth and 60 min ambient air tempering period.

Keywords: drying, fluidized bed dryer, head rice yield, paddy

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Authors: Z. Nurzaireena, K. Azalea, T. Azirawaty, S. Jameela, G. Muralitharan

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The aim of this study is the review of the management practices of sickle cell disease patients during pregnancy, as well as the maternal and neonatal outcome at Ampang Hospital, Selangor. The study consisted of a review of pregnant patients with sickle cell disease under follow up at the Hematology Clinic, Ampang Hospital over the last seven years to assess their management and maternal-fetal outcome. The results of the review show that Ampang Hospital is considered the public hematology centre for sickle cell disease and had successfully managed three pregnancies throughout the last seven years. Patients’ presentations, managements and maternal-fetal outcome were compared and reviewed for academic improvements. All three patients were seen very early in their pregnancy and had been given a regime of folic acid, antibiotics and thrombo-prophylactic drugs. Close monitoring of maternal and fetal well being was done by the hematologists and obstetricians. Among the patients, there were multiple admissions during the pregnancy for either a painful sickle cell bone crisis, haemolysis following an infection and anemia requiring phenotype- matched blood and exchange transfusions. Broad spectrum antibiotics coverage during and infection, hydration, pain management and venous-thrombolism prophylaxis were mandatory. The pregnancies managed to reach near term in the third trimester but all required emergency caesarean section for obstetric indications. All pregnancies resulted in live births with good fetal outcome. During post partum all were nursed closely in the high dependency units for further complications and were discharged well. Post partum follow up and contraception counseling was comprehensively given for future pregnancies. Sickle cell disease is uncommonly seen in the East, especially in the South East Asian region, yet more cases are seen in the current decade due to improved medical expertise and advance medical laboratory technologies. Pregnancy itself is a risk factor for sickle cell patients as increased thrombosis event and risk of infections can lead to multiple crisis, haemolysis, anemia and vaso-occlusive complications including eclampsia, cerebrovasular accidents and acute bone pain. Patients mostly require multiple blood product transfusions thus phenotype-matched blood is required to reduce the risk of alloimmunozation. Emphasizing the risks and complications in preconception counseling and establishing an ultimate pregnancy plan would probably reduce the risk of morbidity and mortality to the mother and unborn child. Early management for risk of infection, thromboembolic events and adequate hydration is mandatory. A holistic approach involving multidisciplinary team care between the hematologist, obstetricians, anesthetist, neonatologist and close nursing care for both mother and baby would ensure the best outcome. In conclusion, sickle cell disease by itself is a high risk medical condition and pregnancy would further amplify the risk. Thus, close monitoring with combine multidisciplinary care, counseling and educating the patients are crucial in achieving the safe outcome.

Keywords: anaemia, haemoglobinopathies, pregnancy, sickle cell disease

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Authors: H. Fuad Usman, M. Rafay Khan Sial, Shahzaib Hamid

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The trend of renewable energy resources has been amplified due to global warming and other environmental related complications in the 21st century. Recent research has very much emphasized on the generation of electrical power through renewable resources like solar, wind, hydro, geothermal, etc. The use of the photovoltaic cell has become very public as it is very useful for the domestic and commercial purpose overall the world. Although a single cell gives the low voltage output but connecting a number of cells in a series formed a complete module of the photovoltaic cells, it is becoming a financial investment as the use of it fetching popular. This also reduced the prices of the photovoltaic cell which gives the customers a confident of using this source for their electrical use. Photovoltaic cell gives the MPPT at single specific point of operation at a given temperature and level of solar intensity received at a given surface whereas the focal point changes over a large range depending upon the manufacturing factor, temperature conditions, intensity for insolation, instantaneous conditions for shading and aging factor for the photovoltaic cells. Two improved algorithms have been proposed in this article for the MPPT. The widely used algorithms are the ‘Incremental Conductance’ and ‘Perturb and Observe’ algorithms. To extract the maximum power from the source to the load, the duty cycle of the convertor will be effectively controlled. After assessing the previous techniques, this paper presents the improved and reformed idea of harvesting maximum power point from the photovoltaic cells. A thoroughly go through of the previous ideas has been observed before constructing the improvement in the traditional technique of MPP. Each technique has its own importance and boundaries at various weather conditions. An improved technique of implementing the use of both ‘Perturb and Observe’ and ‘Incremental Conductance’ is introduced.

Keywords: duty cycle, MPPT (Maximum Power Point Tracking), perturb and observe (P&O), photovoltaic module

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Authors: Ghada Esheba, Ebtisam Aljerayan, Afnan Al-Ghamdi, Atheer Alsharif, Hanan alzahrani

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Background: Primary ovarian neoplasms comprise a heterogeneous group of tumors of three main subtypes: surface epithelial, germ cell, and sex cord-stromal. The wide morphological variation within and between these groups can result in diagnostic difficulties. Gonadal sex cord-stromal tumors (SCST) represent one of the most heterogeneous categories of human neoplasms, because they may contain various combinations of different gonadal sex cord and stromal element. Aim: The aim of this work is to highlight the clinicopathological characteristics of SCST and to assess the value of alpha-inhibin and calretinin in the distinction between SCST and their mimics. Material and methods: This study was carried out on 100 cases using full tissue sections; 70 cases were SCST and 30 cases were histological mimics of SCST. The cases were studied using immunohistochemically using alpha-inhibin. In addition, an ovarian tissue microarray containing 170 benign and malignant ovarian neoplasms was also studied immunohistochemically for calretinin expression. The ovarian microarray included 14 SCST, 59 ovarian serous borderline tumors, 17 mucinous borderline tumors, 10 mucinous adenocarcinomas, 32 endometrioid adenocarcinomas, 34 clear cell carcinomas, and 4 germ cell tumors. Results: 99% of SCST examined using full tissue sections exhibited positive cytoplasmic staining for inhibin. On the contrary, only 7% of the histological mimics (P value < 0.0001). 86% of SCST in the tissue microarray were positive for calretinin with nuclear and/or cytoplasmic staining compared to only 7% of the other tumor types (P value < 0.0001). Conclusions: SCST have characteristic clinicopathological and immunohistochemical features and their recognition is crucial for proper diagnosis and treatment. Alpha-inhibin and calretinin are of great help in the diagnosis of sex cord-stromal tumors.

Keywords: calretinin, granulosa cell tumor, inhibin, sex cord-stromal tumors

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Authors: Mustafa M. Donma, Orkide Donma, Murat Aydin, Muhammet Demirkol, Burcin Nalbantoglu, Aysin Nalbantoglu, Birol Topcu

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Childhood obesity is an ever increasing global health problem, affecting both developed and developing countries. Accurate evaluation of obesity in children requires difficult and detailed investigation. In our study, obesity in children was evaluated using new body fat ratios and indices. Assessment of anthropometric measurements, as well as some ratios, is important because of the evaluation of gender differences particularly during the late periods of obesity. A total of 239 children; 168 morbid obese (MO) (81 girls and 87 boys) and 71 normal weight (NW) (40 girls and 31 boys) children, participated in the study. Informed consent forms signed by the parents were obtained. Ethics Committee approved the study protocol. Mean ages (years)±SD calculated for MO group were 10.8±2.9 years in girls and 10.1±2.4 years in boys. The corresponding values for NW group were 9.0±2.0 years in girls and 9.2±2.1 years in boys. Mean body mass index (BMI)±SD values for MO group were 29.1±5.4 kg/m² and 27.2±3.9 kg/m² in girls and boys, respectively. These values for NW group were calculated as 15.5±1.0 kg/m² in girls and 15.9±1.1 kg/m² in boys. Groups were constituted based upon BMI percentiles for age-and-sex values recommended by WHO. Children with percentiles >99 were grouped as MO and children with percentiles between 85 and 15 were considered NW. The anthropometric measurements were recorded and evaluated along with the new ratios such as trunk-to-appendicular fat ratio, as well as indices such as Index-I and Index-II. The body fat percent values were obtained by bio-electrical impedance analysis. Data were entered into a database for analysis using SPSS/PASW 18 Statistics for Windows statistical software. Increased waist-to-hip circumference (C) ratios, decreased head-to-neck C, height ‘to’ ‘two’-‘to’-waist C and height ‘to’ ‘two’-‘to’-hip C ratios were observed in parallel with the development of obesity (p≤0.001). Reference value for height ‘to’ ‘two’-‘to’-hip ratio was detected as approximately 1.0. Index-II, based upon total body fat mass, showed much more significant differences between the groups than Index-I based upon weight. There was not any difference between trunk-to-appendicular fat ratios of NW girls and NW boys (p≥0.05). However, significantly increased values for MO girls in comparison with MO boys were observed (p≤0.05). This parameter showed no difference between NW and MO states in boys (p≥0.05). However, statistically significant increase was noted in MO girls compared to their NW states (p≤0.001). Trunk-to-appendicular fat ratio was the only fat-based parameter, which showed gender difference between NW and MO groups. This study has revealed that body ratios and formula based upon body fat tissue are more valuable parameters than those based on weight and height values for the evaluation of morbid obesity in children.

Keywords: anthropometry, childhood obesity, gender, morbid obesity

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Authors: Ishani Das, Avinash Padhi, Sitabja Mukherjee, Santosh Kar, Avinash Sonawane

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Activation of cell mediated and humoral immune responses to Mycobacterium tuberculosis (Mtb) are critical for protection. Herein, we show that mice immunized with Mtb lipid bound chitosan nanoparticles(NPs) induce secretion of prominent Th1 and Th2 cytokines in lymph node and spleen cells, and also induced significantly higher levels of IgG, IgG1, IgG2 and IgM in comparison to control mice measured by ELISA. Furthermore, significantly enhanced γδ-T cell activation was observed in lymph node cells isolated from mice immunized with Mtb lipid coated chitosan-NPs as compared to mice immunized with chitosan-NPs alone or Mtb lipid liposomes through flow cytometric analysis. Also, it was observed that in comparison to CD8+ cells, significantly higher CD4+ cells were present in both the lymph node and spleen cells isolated from mice immunized with Mtb lipid coated chitosan NP. In conclusion, this study represents a promising new strategy for efficient delivery of Mtb lipids using chitosan NPs to trigger enhanced cell mediated and antibody response against Mtb lipids.

Keywords: antibody response, chitosan nanoparticles, cytokines, mycobacterium tuberculosis lipids

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Authors: Subham Ghosh, Arnab Nandi

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Background: The proliferation of Wireless Body Area Networks (WBAN) and Internet of Things (IoT) applications demonstrates the potential for continuous monitoring of physical changes in the body. These technologies are vital for health monitoring tasks, such as identifying coughing and breathing activities, which are necessary for disease diagnosis and management. Monitoring activities such as coughing and deep breathing can provide valuable insights into a variety of medical issues. Wearable radio-based antenna sensors, which are lightweight and easy to incorporate into clothing or portable goods, provide continuous monitoring. This mobility gives it a substantial advantage over stationary environmental sensors like as cameras and radar, which are constrained to certain places. Furthermore, using compressive techniques provides benefits such as reduced data transmission speeds and memory needs. These wearable sensors offer more advanced and diverse health monitoring capabilities. Methodology: This study analyzes the feasibility of using a semi-flexible antenna operating at 2.4 GHz (ISM band) and positioned around the neck and near the mouth to identify three activities: coughing, deep breathing, and idleness. A vector network analyzer (VNA) is used to collect time-varying complex reflection coefficient data from perturbed antenna near field. The reflection coefficient (S11) conveys nuanced information caused by simultaneous variations in the near-field radiation of three activities across time. The signatures are sparsely represented by Gaussian windowed Gabor spectrograms. The antenna is biocompatible in terms of specific absorption rate (SAR). The sparsely represented Gabor spectrogram pictures are fed into a lightweight deep learning (DL) model for feature extraction and classification. Two antenna locations are investigated in order to determine the most effective localization for three different activities. Findings: Cross-validation techniques were used on data from both locations. Due to the complex form of the recorded S11, separate analyses and assessments were performed on the magnitude, phase, and their combination. The combination of magnitude and phase fared better than the separate analyses. Various sliding window sizes, ranging from 1 to 5 seconds, were tested to find the best window for activity classification. It was discovered that a neck-mounted design was effective at detecting the three unique behaviors.

Keywords: activity recognition, antenna, deep-learning, time-frequency

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Authors: Mai M. Farid, Mona El-Shabrawy, Sameh R. Hussein, Ahmed Elkhateeb, El-Said S. Abdel-Hameed, Mona M. Marzouk

Abstract:

Asphodelus aestivus Brot. Is a wild plant distributed in Egypt and is considered one of the five Asphodelus spp. from the family Asphodelaceae; it grows in dry grasslands and on rocky or sandy soil. The chemical components of A. aestivus flowers extract were analyzed using different chromatographic and spectral techniques and led to the isolation of two anthraquinones identified as emodin and emodin-O-glucoside. In addition to, five flavonoid compounds;kaempferol,Kaempferol-3-O-glucoside,Apigenin-6-C-glucoside-7-O-glucoside (Saponarine), luteolin 7-O-β-glucopyranoside, Isoorientin-O-malic acid which is a new compound in nature. The LC-ESI-MS/MS analysis of the flower extract of A. aestivus led to the identification of twenty- two compounds characterized by the presence of flavones, flavonols, and flavone C-glycosides. While GC/MS analysis led to the identification of 24 compounds comprising 98.32% of the oil, the major components of the oil were 9, 12, 15-Octadecatrieoic acid methyl ester 28.72%, and 9, 12-Octadecadieroic acid (Z, Z)-methyl ester 19.96%. In vitro cytotoxic activity of the aqueous methanol extract of A. aestivus flowers against HEPG2, HCT-116, MCF-7, and A549 culture was examined and showed moderate inhibition (62.3±1.1)% on HEPG2 cell line followed by (36.8±0.2)% inhibition on HCT-116 and a weak inhibition (5.7± 0.0.2) on MCF-7 cell line followed by (4.5± 0.4) % inhibition on A549 cell line and this is considered the first cytotoxic report of A. aestivus flowers.

Keywords: Anthraquinones, Asphodelus aestivus, Cytotoxic activity, Flavonoids, LC-ESI-MS/MS

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Authors: Hammad Khan, Wookeun Bae

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Major concerns regarding nitritation of low-strength ammonium wastewaters include low ammonium loading rates (usually below 0.2 kg/m3-d) and uncertainty about long-term stability of the process. The purpose of this study was to test a sequencing batch reactor (SBR) filled with cell-immobilized polyethylene glycol (PEG) pellets to see if it could achieve efficient and stable nitritation under various environmental conditions. SBR was fed with synthetic ammonium wastewater of 30±2 mg-N/L and pH: 8±0.05, maintaining the dissolved oxygen concentration of 1.7±0.2 mg/L and the temperature at 30±1oC. The reaction was easily converted to partial nitrification mode within a month by feeding relatively high ammonium substrate (~100 mg-N/L) in the beginning. We observed stable nitritation over 300 days with high ammonium loading rates (as high as ~1.1 kg-N/m3-d), nitrite accumulation rates (mostly over 97%) and ammonium removal rate (mostly over 95%). DO was a major limiting substrate when the DO concentration was below ~4 mg/L and the NH4+-N concentration was above 5 mg/L, giving almost linear increase in the ammonium oxidation rate with the bulk DO increase. Low temperatures mainly affected the reaction rate, which could be compensated for by increasing the pellet volume (i.e. biomass). Our results demonstrated that an SBR filled with small cell-immobilized PEG pellets could achieve very efficient and stable nitritation of a low-strength ammonium wastewater.

Keywords: ammonium loading rate (ALR), cell-immobilization, long-term nitritation, sequencing batch reactor (SBR), sewage treatment

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Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang

Abstract:

Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).

Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells

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Authors: Yuan-Hao Lee, Chia-Wei Lee, Ming-Fang Lin, Tzu-Huei Wu, Chih-Hsiang Ko, Wing P. Chan

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Computed tomography (CT) scan of the head is a good method for investigating cranial lesions. However, radiation-induced oxidative stress can be accumulated in the eyes and promote carcinogenesis and cataract. In this regard, we aimed to protect the eyes with barium sulfate shield(s) during CT scans and investigate the resultant image quality and radiation dose to the eye. Patients who underwent health examinations were selectively enrolled in this study in compliance with the protocol approved by the Ethics Committee of the Joint Institutional Review Board at Taipei Medical University. Participants’ brains were scanned with a water-based marker simultaneously by a multislice CT scanner (SOMATON Definition Flash) under a fixed tube current-time setting or automatic tube current modulation (TCM). The lens dose was measured by Gafchromic films, whose dose response curve was previously fitted using thermoluminescent dosimeters, with or without barium sulfate or bismuth-antimony shield laid above. For the assessment of image quality CT images at slice planes that exhibit the interested regions on the zygomatic, orbital and nasal bones of the head phantom as well as the water-based marker were used for calculating the signal-to-noise and contrast-to-noise ratios. The application of barium sulfate and bismuth-antimony shields decreased 24% and 47% of the lens dose on average, respectively. Under topogram-based TCM, the dose saving power of bismuth-antimony shield was mitigated whereas that of barium sulfate shield was enhanced. On the other hand, the signal-to-noise and contrast-to-noise ratios of DSCT images were decreased separately by barium sulfate and bismuth-antimony shield, resulting in an overall reduction of the CNR. In contrast, the integration of topogram-based TCM elevated signal difference between the ROIs on the zygomatic bones and eyeballs while preferentially decreasing the signal-to-noise ratios upon the use of barium sulfate shield. The results of this study indicate that the balance between eye exposure and image quality can be optimized by combining eye shields with topogram-based TCM on the multislice scanner. Eye shielding could change the photon attenuation characteristics of tissues that are close to the shield. The application of both shields on eye protection hence is not recommended for seeking intraorbital lesions.

Keywords: computed tomography, barium sulfate shield, dose saving, image quality

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Authors: Shen Jian, Hu Jie, Zhu Guo Niu, Peng Ying Hong

Abstract:

There are two issues exited in the traditional visual prosthesis: lacking systematic method and the low level of humanization. To tackcle those obstacles, a visual prosthesis design method based on biologically inspired design is proposed. Firstly, a constrained FBS knowledge cell model is applied to construct the functional model of visual prosthesis in biological field. Then the clustering results of engineering domain are ob-tained with the use of the cross-domain knowledge cell clustering algorithm. Finally, a prototype system is designed to support the bio-logically inspired design where the conflict is digested by TRIZ and other tools, and the validity of the method is verified by the solution scheme

Keywords: knowledge-based engineering, visual prosthesis, biologically inspired design, biomedical engineering

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Authors: Yik Sin Chan, Nam Weng Sit, Fook Yee Chye, van Ofwegen Leen, de Voogd Nicole, Kong Soo Khoo

Abstract:

Chikungunya fever is an arboviral disease transmitted by the Aedes mosquitoes. It has resulted in epidemics of the disease in tropical countries in the Indian Ocean and South East Asian regions. The recent spread of this disease to the temperate countries such as France and Italy, coupled with the absence of vaccines and effective antiviral drugs make chikungunya fever a worldwide health threat. This study aims to investigate the anti-chikungunya virus activity of selected marine organism samples collected from Malaysian coastal areas, including seaweeds (Caulerpa racemosa, Caulerpa sertularioides and Kappaphycus alvarezii), a soft coral (Lobophytum microlobulatum) and a sponge (Spheciospongia vagabunda). Following lyophilization (oven drying at 40C for K. alvarezii) and grinding to powder form, each sample was subjected to sequential solvent extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water in order to extract bioactive compounds. The antiviral activity was evaluated using monkey kidney epithelial (Vero) cells infected with the virus (multiplicity of infection=1). The cell viability was determined by Neutral Red uptake assay. 70% of the 30 extracts showed weak inhibitory activity with cell viability ≤30%. Seven of the extracts exhibited moderate inhibitory activity (cell viability: 31%-69%). These were the chloroform, ethyl acetate, ethanol and methanol extracts of C. racemosa; chloroform and ethyl acetate extracts of L. microlobulatum; and the chloroform extract of C. sertularioides. Only the hexane and ethanol extracts of L. microlobulatum showed strong inhibitory activity against the virus, resulting in cell viabilities (mean±SD; n=3) of 73.3±2.6% and 79.2±0.9%, respectively. The corresponding mean 50% effective concentrations (EC50) for the extracts were 14.2±0.2 and 115.3±1.2 µg/mL, respectively. The ethanol extract of the soft coral L. microlobulatum appears to hold the most promise for further characterization of active principles as it possessed greater selectivity index (SI>5.6) compared to the hexane extract (SI=2.1).

Keywords: antiviral, seaweed, sponge, soft coral, vero cell

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Authors: Cuili Yang, Chengcao Sun, Ruilin Xue, Yongyong Xi, Liang Wang, Dejia Li

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Backgrounds: Previous studies showed that Sirt1 plays an important role in C2C12 myoblast cell proliferation, but the mechanism(s) involved in this process remains unclear. This work was undertaken to determine if Myostatin participates in the regulation of C2C12 proliferation by Sirt1. Methods: We administrated the Sirt1 activator resveratrol, inhibitor Nicotinamide (NAM) and Myostatin inhibitor SB431542 on C2C12 myoblast cells. Cell viability was evaluated by CCK8 assay. The expression of Sirt1 and MyoD were detected by qRT-PCR. Utilizing western blot to determinate the expression of myostatin, P107 and p-P107. Results: Our results showed that resveratrol promoted the proliferation of C2C12 myoblast cells, while NAM suppressed the proliferation of C2C12 myoblast cells; SB431542 promoted the proliferation of C2C12 myoblast cells and attenuated the inhibition effect of NAM on C2C12 myoblast cells proliferation; Resveratrol can significantly increase the expression of Sirt1 and MyoD, decrease the expression of Myostatin, while NAM can significantly down-regulate the expression of Sirt1, MyoD and the phosphorylation of P107(p-P107), but up-regulate the expression of Myostatin and the protein P107; SB431542 can significantly mitigate the effect of NAM on the expression of MyoD, P107, and p-P107. Conclusions: Taken together, these results indicate that Sirt1 promotes the proliferation of C2C12 myoblast cells via Myostatin signaling pathway.

Keywords: Sirt1, C2C12 cells, proliferation, myostatin signaling pathway

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Authors: Samarjeet Singh, Shubhank Arya, Shubham Chauhan

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In recent years, the rise in carbon emissions and the widespread effects of global warming have brought new energy vehicles into the spotlight. Electric vehicles (EVs) and hydrogen fuel cell vehicles (HFCVs), both producing zero tailpipe emissions, are seen as promising alternatives. This paper examines the working, structural characteristics, and safety designs of EVs and HFCVs, comparing their carbon emissions, charging infrastructure, energy efficiency, and safety features. The analysis reveals that both EVs and HFCVs significantly reduce carbon emissions and enhance safety compared to traditional vehicles, with EVs showing greater emission reductions. Moreover, EVs are advancing more rapidly in terms of charging infrastructure compared to hydrogen energy vehicles. However, HFCVs exhibit lower energy efficiency than EVs. In terms of safety, both types surpass conventional vehicles, though EVs are more prone to overheating and fire hazards due to battery design issues. Current research suggests that EV technology and its supporting infrastructure are more comprehensive, cost-effective, and efficient in reducing carbon emissions. With continued investment in the development of new energy vehicles and potential advancements in hydrogen energy production, the future for HFCVs appears promising. The paper also expresses optimism for innovative solutions that could accelerate the growth of hydrogen energy vehicles.

Keywords: electric vehicles, fuel cell electric vehicles, automotive engineering, energy transition

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Authors: Mahdiye Khorasani, Arjun Vijay, Ali Mashayekh, Christian Trapp

Abstract:

The performance of Proton Exchange Membrane Fuel Cell (PEMFC) is highly dependent on the pressure and mass flow of media (Hydrogen and air) throughout the cells and the stack. Higher pressure, on the one hand, results in higher output power of the stack but, on the other hand, increases the electrical power demand of the compressor. In this work, a simulation model of a PEMFC system for vehicle and airplane applications is developed. With this new model, the effect of different pressures and air mass flow rates are investigated to discover the optimum operating point in a PEMFC system, and innovative operation strategies are implemented to optimize reactants flow while minimizing electrical power demand of the compressor for optimum performance. Additionally, a fuel cell system test bench is set up, which contains not only all the auxiliary components for conditioning the gases, reactants, and flows but also a dynamic titling table for testing different orientations of the stack to simulate the flight conditions during take-off and landing and off-road-vehicle scenarios. The results of simulation will be tested and validated on the test bench for future works.

Keywords: air mass flow effect, optimization of operation, pressure effect, PEMFC system, PEMFC system simulation

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Authors: M. Aouimeur

Abstract:

The environmental risks of a dam and particularly the security in the Valley downstream of it,, is a very complex problem. Integrated management and risk-sharing become more and more indispensable. The definition of "vulnerability “concept can provide assistance to controlling the efficiency of protective measures and the characterization of each valley relatively to the floods's risk. Security can be enhanced through the integrated land management. The social sciences may be associated to the operational systems of civil protection, in particular warning networks. The passage of extreme floods in the site of the dam causes the rupture of this structure and important damages downstream the dam. The river bed could be damaged by erosion if it is not well protected. Also, we may encounter some scouring and flooding problems in the downstream area of the dam. Therefore, the protection of the dam is crucial. It must have an energy dissipator in a specific place. The basin of dissipation plays a very important role for the security of the dam and the protection of the environment against floods downstream the dam. It allows to dissipate the potential energy created by the dam with the passage of the extreme flood on the weir and regularize in a natural manner and with more security the discharge or elevation of the water plan on the crest of the weir, also it permits to reduce the speed of the flow downstream the dam, in order to obtain an identical speed to the river bed. The problem of the dimensioning of a classic dissipation basin is in the determination of the necessary parameters for the dimensioning of this structure. This communication presents a simple graphical method, that is fast and complete, and a methodology which determines the main features of the hydraulic jump, necessary parameters for sizing the classic dissipation basin. This graphical method takes into account the constraints imposed by the reality of the terrain or the practice such as the one related to the topography of the site, the preservation of the environment equilibrium and the technical and economic side.This methodology is to impose the loss of head DH dissipated by the hydraulic jump as a hypothesis (free design) to determine all the others parameters of classical dissipation basin. We can impose the loss of head DH dissipated by the hydraulic jump that is equal to a selected value or to a certain percentage of the upstream total head created by the dam. With the parameter DH+ =(DH/k),(k: critical depth),the elaborate graphical representation allows to find the other parameters, the multiplication of these parameters by k gives the main characteristics of the hydraulic jump, necessary parameters for the dimensioning of classic dissipation basin.This solution is often preferred for sizing the dissipation basins of small concrete dams. The results verification and their comparison to practical data, confirm the validity and reliability of the elaborate graphical method.

Keywords: dimensioning, energy dissipation basin, hydraulic jump, protection of the environment

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Authors: Shaimaa Eltanani, Thangal Yumnamcha, Ahmed S. Ibrahim

Abstract:

Purpose: Mitochondria dysfunction is central to breaking the barrier integrity of retinal endothelial cells (RECs) in various blinding eye diseases such as diabetic retinopathy and retinopathy of prematurity. Therefore, we aimed to dissect the role of different mitochondrial components, specifically, those of oxidative phosphorylation (OxPhos), in maintaining the barrier function of RECs. Methods: Electric cell-substrate impedance sensing (ECIS) technology was used to assess in real-time the role of different mitochondrial components in the total impedance (Z) of human RECs (HRECs) and its components; the capacitance (C) and the total resistance (R). HRECs were treated with specific mitochondrial inhibitors that target different steps in OxPhos: Rotenone for complex I; Oligomycin for ATP synthase; and FCCP for uncoupling OxPhos. Furthermore, data were modeled to investigate the effects of these inhibitors on the three parameters that govern the total resistance of cells: cell-cell interactions (Rb), cell-matrix interactions (α), and cell membrane permeability (Cm). Results: Rotenone (1 µM) produced the greatest reduction in the Z, followed by FCCP (1 µM), whereas no reduction in the Z was observed after the treatment with Oligomycin (1 µM). Following this further, we deconvoluted the effect of these inhibitors on Rb, α, and Cm. Firstly, rotenone (1 µM) completely abolished the resistance contribution of Rb, as the Rb became zero immediately after the treatment. Secondly, FCCP (1 µM) eliminated the resistance contribution of Rb only after 2.5 hours and increased Cm without considerable effect on α. Lastly, Oligomycin had the lowest impact among these inhibitors on Rb, which became similar to the control group at the end of the experiment without noticeable effects on Cm or α. Conclusion: These results demonstrate differential roles for complex I, complex V, and coupling of OxPhos in maintaining the barrier functionality of HRECs, in which complex I being the most important component in regulating the barrier functionality and the spreading behavior of HRECs. Such differences can be used in investigating gene expression as well as for screening selective agents that improve the functionality of complex I to be used in the therapeutic approach for treating REC-related retinal diseases.

Keywords: human retinal endothelial cells (hrecs), rotenone, oligomycin, fccp, oxidative phosphorylation, oxphos, capacitance, impedance, ecis modeling, rb resistance, α resistance, and barrier integrity

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Authors: Janneth Gonzalez, Marco Avila, George Barreto

Abstract:

GRP78 participates in multiple functions in the cell during normal and pathological conditions, controlling calcium homeostasis, protein folding and unfolded protein response. GRP78 is located in the endoplasmic reticulum, but it can change its location under stress, hypoxic and apoptotic conditions. NF-κB represents the keystone of the inflammatory process and regulates the transcription of several genes related with apoptosis, differentiation, and cell growth. The possible relationship between GRP78-NF-κB could support and explain several mechanisms that may regulate a variety of cell functions, especially following brain injuries. Although several reports show interactions between NF-κB and heat shock proteins family members, there is a lack of information on how GRP78 may be interacting with NF-κB, and possibly regulating its downstream activation. Therefore, we assessed the computational predictions of the GRP78 (Chain A) and NF-κB complex (IkB alpha and p65) protein-protein interactions. The interaction interface of the docking model showed that the amino acids ASN 47, GLU 215, GLY 403 of GRP78 and THR 54, ASN 182 and HIS 184 of NF-κB are key residues involved in the docking. The electrostatic field between GRP78-NF-κB interfaces and molecular dynamic simulations support the possible interaction between the proteins. In conclusion, this work shed some light in the possible GRP78-NF-κB complex indicating key residues in this crosstalk, which may be used as an input for better drug design strategy targeting NF-κB downstream signaling as a new therapeutic approach following brain injuries.

Keywords: computational biology, protein interactions, Grp78, bioinformatics, molecular dynamics

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