Search results for: stress urinary incontinence (SU
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4149

Search results for: stress urinary incontinence (SU

2259 The Effectiveness of Laughing Qigong for Women with Breast Cancer in Community

Authors: Chueh Chang, Chia-jung Hsieh, Fu-yu Yu, Yu-Hwa Lin

Abstract:

Background:The majority of women diagnosed with breast cancer undergo treatment involving surgery and radiotherapy or chemotherapy, or both. With these major advances in breast cancer management, many patients still have to deal with short or long-term side effects and psychological distress related to the disease and treatment, which have a substantial impact on their quality of life. The Laughing Qigong Program (LQP) is an interactive laughter program that combines the physical and physiological benefits of laughter with the mental benefits of Chinese qigong. Purpose: In order to improve the quality of life for breast cancer women in the community as well as echoing the WHO 2004 “Promoting Mental Health” for every one. This study focused on how to promote the positive mental health for women of breast cancer through the “laughter program” in Taiwan. During the presentation, how to practice Laughing Qigong will be demonstrated. Method: Using nonequivalent pretest-posttest design, ix-one breast cancer patients were volunteered to enroll in this study from the Taiwan Breast Cancer Alliance (TBCA). Thirty patients were assigned to the experimental group and the other 31 patients were assigned to the control group. The women who were assigned to the experimental group received laughter program one hour per session, once a week, totally 12 sessions. All subjects were tested before and after the intervention on the following: Self-Esteem scale (RSE), Face Scale (FS), Anxiety and pain experience were measured as psychological markers; saliva cortisol (CS) as an immunological marker; blood pressure (BP), heart rate (HR),and heart rate variability (HRV) as physiological markers of the body’s response to stress. Results: After comparing the experimental and control groups, the results revealed that those breast cancer women with “laughing program” their sense of humor were improved, less uncomfortable on self report physical conditions, more positive attitudes toward stress management by using laughter, and had emotional improvement according to the face scale.

Keywords: mental health promotion, breast cancer, laughing Qigong, women

Procedia PDF Downloads 489
2258 Role of Onion Extract for Neuro-Protection in Experimental Stroke Model

Authors: Richa Shri, Varinder Singh, Kundan Singh Bora, Abhishek Bhanot, Rahul Kumar, Amit Kumar, Ravinder Kaur

Abstract:

The term ‘neuroprotection’ means preserving/salvaging function and structure of neurons. Neuroprotection is an adjunctive treatment option for neurodegenerative disorders. Oxidative stress is considered a major culprit in neurodegenerative disorders; hence, management strategies include use of antioxidants. Our search for a neuroprotective agent began with Allium cepa L. or onions, (family Amaryllidaceae) - a potent antioxidant. We have investigated the neuroprotective potential of onions in experimental models of ischemic stroke, diabetic neuropathy, neuropathic pain, and dementia. In pre and post-ischemic stroke model, the methanol extract of outer scales of onion bulbs (MEOS) prevented memory loss and motor in-coordination; reduced oxidative stress and cerebral infarct size. This also prevented and ameliorated diabetic neuropathy in mice. The MEOS was fractionated to yield a flavonoid rich fraction (FRF) that successfully reversed ischemia-reperfusion induced neuronal damage, thereby demonstrating that the flavonoids are responsible for the activity. The FRF effectively ameliorated chronic constriction induced neuropathic pain in rats. The FRF was subjected to bioactivity-guided fractionated. It was seen that FRF is more effective as compared to the isolated components probably due to synergism among the constituents (i.e., quercetin and quercetin glucosides) in the FRF. The outer scales of onion bulbs have great potential for prevention as well as for treatment of neuronal disorders. Red onions, with higher amounts of flavonoids as compared to the white onions, produced more significant neuroprotection. Thus, the standardized FRF from the waste material of a commonly used vegetable, especially the red variety, may be developed as a valuable neuroprotective agent.

Keywords: Allium cepa, antioxidant activity, flavonoid rich fraction, neuroprotection

Procedia PDF Downloads 152
2257 Possible Endocrinal and Liver Enzymes Toxicities Associated with Long Term Exposure to Benzene in Saudi Arabia

Authors: Faizah Asiri, Mohammed Fathy, Saeed Alghamdi, Nahlah Ayoub, Faisal Asiri

Abstract:

Background: - The strategies for this study were based on the toxic effect of long-term inhalation of Benzene on hormones and liver enzymes and various parameters related to it. The following databases were searched: benzene, hepatotoxic, benzene metabolism, hormones, testosterone, hemotoxic, and prolonged exposure. A systematic strategy is designed to search the literature that links benzene with the multiplicity and different types of intoxication or the medical abbreviations of diseases relevant to benzene exposure. Evidence suggests that getting rid of inhaled gasoline is by exhalation. Absorbed benzene is metabolized by giving phenolic acid as well as meconic acid, followed by urinary excretion of conjugate sulfates and glucuronides. Materials and Methods :- This work was conducted in the Al-Khadra laboratory in Taif 2020/2021 and aimed to measure some of the possible endocrinal and liver toxicities associated with benzene's long-term exposure in Saudi Arabia at the station workers who are considered the most exposed category to gasoline. One hundred ten station workers were included in this study. They were divided into four patient groups according to the chronic exposure rate to benzene, one control group, and three other groups of exposures. As follows: patient Group 1 (controlled group), patient Group 2 (exposed less than 1y), patient Group 3 (exposed 1-5 y), patient Group 4 (more than 5). Each group is compared with blood sample parameters (ALT, FSH and Testosterone, TSH). Blood samples were drawn from the participants, and statistical tests were performed. Significant change (p≤0.05) was examined compared to the control group. Workers' exposure to benzene led to a significant change in hematological, hormonal, and hepatic factors compared to the control group. Results:- The results obtained a relationship between long-term exposure to benzene and a decrease in the level of testosterone and FSH hormones, including that it poses a toxic risk in the long term (p≤0.05) when compared to the control. We obtained results confirming that there is no significant coloration between years of exposure and TSH level (p≤0.05) when compared to the control. Conclusion:- We conclude that some hormones and liver enzymes are affected by chronic doses of benzene through inhalation after our study was on the group most exposed to benzene, which is gas station workers.

Keywords: toxicities, benzene, hormones, station workers

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2256 Chemical Aging of High-Density Polyethylene (HDPE-100) in Interaction with Aggressive Environment

Authors: Berkas Khaoula, Chaoui Kamel

Abstract:

Polyethylene (PE) pipes are one of the best options for water and gas transmission networks. The main reason for such a choice is its high-quality performance in service conditions over long periods of time. PE pipes are installed in contact with different soils having various chemical compositions with confirmed aggressiveness. As a result, PE pipe surfaces undergo unwanted oxidation reactions. Usually, the polymer mixture is designed to include some additives, such as anti-oxidants, to inhibit or reduce the degradation effects. Some other additives are intended to increase resistance to the ESC phenomenon associated with polymers (ESC: Environmental Stress Cracking). This situation occurs in contact with aggressive external environments following different contaminations of soil, groundwater and transported fluids. In addition, bacterial activity and other physical or chemical media, such as temperature and humidity, can play an enhancing role. These conditions contribute to modifying the PE pipe structure and degrade its properties during exposure. In this work, the effect of distilled water, sodium hypochlorite (bleach), diluted sulfuric acid (H2SO4) and toluene-methanol (TM) mixture are studied when extruded PE samples are exposed to those environments for given periods. The chosen exposure periods are 7, 14 and 28 days at room temperature and in sealed glass containers. Post-exposure observations and ISO impact tests are presented as a function of time and chemical medium. Water effects are observed to be limited in explaining such use in real applications, whereas the changes in TM and acidic media are very significant. For the TM medium, the polymer toughness increased drastically (from 15.95 kJ/m² up to 32.01 kJ/m²), while sulfuric acid showed a steady augmentation over time. This situation may correspond to a hardening phenomenon of PE increasing its brittleness and its ability for structural degradation because of localized oxidation reactions and changes in crystallinity.

Keywords: polyethylene, toluene-methanol mixture, environmental stress cracking, degradation, impact resistance

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2255 Fracture and Fatigue Crack Growth Analysis and Modeling

Authors: Volkmar Nolting

Abstract:

Fatigue crack growth prediction has become an important topic in both engineering and non-destructive evaluation. Crack propagation is influenced by the mechanical properties of the material and is conveniently modelled by the Paris-Erdogan equation. The critical crack size and the total number of load cycles are calculated. From a Larson-Miller plot the maximum operational temperature can for a given stress level be determined so that failure does not occur within a given time interval t. The study is used to determine a reasonable inspection cycle and thus enhances operational safety and reduces costs.

Keywords: fracturemechanics, crack growth prediction, lifetime of a component, structural health monitoring

Procedia PDF Downloads 49
2254 Role of Autophagic Lysosome Reformation for Cell Viability in an in vitro Infection Model

Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

Abstract:

Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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2253 Fathers’ Depression and its Relationship with Mothers’ Depression During Postpartum Period

Authors: Fatemeh Abdollahi, Munn-Sann Lye, Jamshid Yazdani Charati, Mehran Zarghami

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Fathers are at risk of depression during the postpartum period. Some studies have been reported maternal depression is the key predictor of paternal postpartum depression (PPD). This study aimed to estimate the prevalence and predictors of parental PPD and its association with maternal PPD. In a cross-sectional study, via a stratified random and convenience sampling method, participants referring to health centers during 2-8 weeks postpartum were recruited from March to October 2017. Paternal PPD and its relation to maternal PPD and other related factors were assessed using multiple logistic regression. Participants were 591 literate couples who referred to Mazandaran province primary health centers during to study period. Couples were screened for depression using Edinburgh Postnatal Depression Scale (EPDS). Fathers provided information on socio-demographic characteristics, life events, neonatal stressor, perceived stress (Perceived Stress Scale), social support (Multidimensional Scale of Perceived Social Support), and general health status using General Health Questionnaire (GHQ) as well. Data on mothers ‘demographic characteristics and obstetrics factors was also gathered. Overall, 93 fathers (15.7%) and 188 mothers (31.8%) reported depressive symptoms above the cut-off EPDS score of 12. In the multiple logistic regression model, older age [OR=1.20, (95%CI: 1.05- 1.36)], maternal depressive symptoms [OR=1.15, (95%CI: 1.04-1.27)], higher GHQ scores [OR=1.21, (95%CI: 1.11-1.33)] and increased recent life events [OR=1.42, (95%CI: 1.01-1.2.00)] were related to paternal PPD. A significant inverse association was found between number of children and paternal PPD [OR=0.20, (95%CI: 0.07-0.53)]. Depressive symptoms, especially in first-time fathers following the birth of a child, are not uncommon. Maternal depressive symptoms and paternal well-being were strong predictors of parental PPD. Creating opportunities for men to access special health care services, parental education to help adapting to parenthood, screening programs, and psychiatric/psychosocial interventions to decrease the suffering of depression for both depressed parents are recommended.

Keywords: depression, men, postpartum, risk factors, women

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2252 The h3r Antagonist E159 Alleviates Neuroinflammation and Autistic-Like Phenotypes in BTBR T+ tf/J Mouse Model of Autism

Authors: Shilu Deepa Thomas, P. Jayaprakash, Dorota Łazewska, Katarzyna Kieć-Kononowicz, B. Sadek

Abstract:

A large body of evidence suggests the involvement of cognitive impairment, increased levels of inflammation and oxidative stress in the pathogenesis of autism spectrum disorder (ASD). ASD commonly coexists with psychiatric conditions like anxiety and cognitive challenges, and individuals with ASD exhibit significant levels of inflammation and immune system dysregulation. Previous Studies have identified elevated levels of pro-inflammatory markers such as IL-1β, IL-6, IL-2 and TNF-α, particularly in young children with ASD. The current therapeutic options for ASD show limited effectiveness, signifying the importance of exploring an efficient drugs to address the core symptoms. The role of histamine H3 receptors (H3Rs) in memory and the prospective role of H3R antagonists in pharmacological control of neurodegenerative disorders, e.g., ASD, is well-accepted. Hence, the effects of chronic systemic administration of H3R antagonist E159 on autistic-like repetitive behaviors, social deficits, memory and anxiety parameters, as well as neuroinflammation in Black and Tan BRachyury (BTBR) mice, were evaluated using Y maze, Barnes maze, self-grooming, open field and three chamber social test. E159 (2.5, 5 and 10 mg/kg, i.p.) dose-dependently ameliorated repetitive and compulsive behaviors by reducing the increased time spent in self-grooming and improved reduced spontaneous alternation in BTBR mice. Moreover, treatment with E159 attenuated disturbed anxiety levels and social deficits in tested male BTBR mice. Furthermore, E159 attenuated oxidative stress by significantly increasing GSH, CAT, and SOD and decreasing the increased levels of MDA in the cerebellum as well as the hippocampus. In addition, E159 decreased the elevated levels of proinflammatory cytokines (tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and IL-6). The observed results show that H3R antagonists like E159 may represent a promising novel pharmacological strategy for the future treatment of ASD.

Keywords: histamine H3 receptors, antagonist E159, autism, behaviors, mice

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2251 Effect of Green Coffee Bean Extract on Gentamicin Induced Acute Renal Failure in Rats

Authors: Amina Unis, Samah S. El Basateeny, Noha A. H. Nassef

Abstract:

Introduction: Acute Renal Failure (ARF) is one of the most common problems encountered in hospitalized critically ill patients. In recent years great effort has been focused on the introduction of herbal medicine as a novel therapeutic agent for prevention of ARF. Hence, the current study was designed to investigate the effect of Green Coffee Bean Extract (GCBE) on gentamicin induced ARF in rats. Methods: The study was conducted on 60 male rats divided into six equal groups. Group 1 served as normal control group and GCBE was administered for 7 days at a dose of 20 mg/kg/day in group 2 and 40 mg/kg/day in group 3 to test the effect of GCBE on normal kidneys. ARF was induced by a daily intraperitoneal injection of gentamicin (80 mg/kg) for 7 days in group 4 (model group), group 5 (GCBE 20 mg/kg/day) and group 6 (GCBE 20 mg/kg/day). All rats were sacrificed after 7 days and blood was withdrawn for kidney function tests. Kidneys were removed for determination of renal oxidative stress markers and histopathological examination. Results: The present study showed that rats that received oral GCBE for 7 days without induction of ARF showed no significant change in all the assessed parameters in comparison to the normal control group, while rats in the groups that received oral GCBE for 7 days with induction of ARF showed a significant improvement in kidney functions tests (decrease in serum urea, serum creatinine, and blood urea nitrogen) when compared to the ARF model group. Moreover, there was significant amelioration in renal oxidative stress markers (renal malondialdehyde, renal superoxide dismutase) and renal histopathological changes in the GCBE treated groups along induction of ARF when compared to ARF model group. The most significant improvement was reported in the group where GCBE was administered for 7 days in a dose 40 mg/kg/day, along with induction of ARF. Conclusion: GCBE has a potential role in ameliorating renal damage involved in ARF mostly through its antioxidant effect.

Keywords: green coffee bean extract, gentamicin, acute renal failure, pharmacology

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2250 Ability of Bentonite-lactobacillus Rhamnosus GAF06 Mixture to Mitigate Aflatoxin M1 Damages in Balb/C Mice

Authors: Amina Aloui, Jalila Ben Salah-Abbès, Abdellah Zinedine, Amar Riba, Noel Durand, Catherine Brabet, Didier Montet, Samir Abbès

Abstract:

Mycotoxin contamination of food and feed-isa globaconcern, both economically and for public health. Aflatoxin M1 (AFM1) is the principal hydroxylated metabolite of aflatoxin B1. It is frequently found in milk and other dairy products. It is responsible for the development of hepatocellular carcinoma and immunotoxic in humans and animals. The reduction of its bioavailabilitybecomesa great demand in order to protect human and animal health. The use of probiotic bacteria and clay are demonstrated to be able to bind AFM1 in vitro. This study aimed to investigate, in vivo, the activity of two-component mixture: L. rhamnosusGAF06 (LR) and bentonite for reducing the oxidative stress and the histological alterationsinduced by AFM1 in the liver andkidneys. For the experiment, male mice were divided into 7 groups (6 mice/group) and treated, orally, by AFM1, alone or in combination with LR and/or bentonite, for 10 days as follows: group 1 control, group 2 treated with LR alone (2.108 CFU/ml), group 3 treated with bentonite alone (1g/kg), group 4 treated with AFM1 alone (100μg/kg), group 5 co-treated with LR+AFM1, group 6 co-treated with bentonite+AFM1, group 7 co-treated with bentonite+LR+AFM1. At the end of the treatment, the mice were sacrificed, and the livers and kidneys were collected for histological assays. Intracellular antioxidant activities and lipid peroxidation were also studied. The results showed that AFM1causeddamage in liver and kidney tissues, being evidence of hepatotoxicity and nephrotoxicity marked by necrotic cells. It increased the MDA level and decreased the antioxidant enzyme activities (SOD) in both organs. In contrast, the co-treatment with AFM1 plus LR and/or bentonitesignificantly improved the hepatic and renal tissues, regulated kidney, and liver antioxidant enzyme activities. This improvement was more remarkable with the administration of LR-bentonite mixture with AFM1.LR and bentonite alone showed to be safe during the treatment. This mixture can be a promising candidate for future applications in biotechnological processes that aimed to detoxify AFM1in food and feed.

Keywords: aflatoxin M1, bentonite, L. rhamnosus GAF06, oxidative stress, prevention

Procedia PDF Downloads 194
2249 Characterization Microstructural Dual Phase Steel for Application In Civil Engineering

Authors: S. Habibi, T. E. Guarcia, A. Megueni, A. Ziadi, L. Aminallah, A. S. Bouchikhi

Abstract:

The characterization of the microstructure of Dual Phase steel in various low-carbon, with a yield stress between 400 and 900 MPa were conducted .In order to assess the mechanical properties of steel, we examined the influence of their chemical compositions interictal and heat treatments (austenite + ferrite area) on their micro structures. In this work, we have taken a number of commercial DP steels, micro structurally characterized and used the conventional tensile testing of these steels for mechanical characterization.

Keywords: characterization, construction in civil engineering, micro structure, tensile DP steel

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2248 Investigation of Delamination Process in Adhesively Bonded Hardwood Elements under Changing Environmental Conditions

Authors: M. M. Hassani, S. Ammann, F. K. Wittel, P. Niemz, H. J. Herrmann

Abstract:

Application of engineered wood, especially in the form of glued-laminated timbers has increased significantly. Recent progress in plywood made of high strength and high stiffness hardwoods, like European beech, gives designers in general more freedom by increased dimensional stability and load-bearing capacity. However, the strong hygric dependence of basically all mechanical properties renders many innovative ideas futile. The tendency of hardwood for higher moisture sorption and swelling coefficients lead to significant residual stresses in glued-laminated configurations, cross-laminated patterns in particular. These stress fields cause initiation and evolution of cracks in the bond-lines resulting in: interfacial de-bonding, loss of structural integrity, and reduction of load-carrying capacity. Subsequently, delamination of glued-laminated timbers made of hardwood elements can be considered as the dominant failure mechanism in such composite elements. In addition, long-term creep and mechano-sorption under changing environmental conditions lead to loss of stiffness and can amplify delamination growth over the lifetime of a structure even after decades. In this study we investigate the delamination process of adhesively bonded hardwood (European beech) elements subjected to changing climatic conditions. To gain further insight into the long-term performance of adhesively bonded elements during the design phase of new products, the development and verification of an authentic moisture-dependent constitutive model for various species is of great significance. Since up to now, a comprehensive moisture-dependent rheological model comprising all possibly emerging deformation mechanisms was missing, a 3D orthotropic elasto-plastic, visco-elastic, mechano-sorptive material model for wood, with all material constants being defined as a function of moisture content, was developed. Apart from the solid wood adherends, adhesive layer also plays a crucial role in the generation and distribution of the interfacial stresses. Adhesive substance can be treated as a continuum layer constructed from finite elements, represented as a homogeneous and isotropic material. To obtain a realistic assessment on the mechanical performance of the adhesive layer and a detailed look at the interfacial stress distributions, a generic constitutive model including all potentially activated deformation modes, namely elastic, plastic, and visco-elastic creep was developed. We focused our studies on the three most common adhesive systems for structural timber engineering: one-component polyurethane adhesive (PUR), melamine-urea-formaldehyde (MUF), and phenol-resorcinol-formaldehyde (PRF). The corresponding numerical integration approaches, with additive decomposition of the total strain are implemented within the ABAQUS FEM environment by means of user subroutine UMAT. To predict the true stress state, we perform a history dependent sequential moisture-stress analysis using the developed material models for both wood substrate and adhesive layer. Prediction of the delamination process is founded on the fracture mechanical properties of the adhesive bond-line, measured under different levels of moisture content and application of the cohesive interface elements. Finally, we compare the numerical predictions with the experimental observations of de-bonding in glued-laminated samples under changing environmental conditions.

Keywords: engineered wood, adhesive, material model, FEM analysis, fracture mechanics, delamination

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2247 Condition Monitoring for Controlling the Stability of the Rotating Machinery

Authors: A. Chellil, I. Gahlouz, S. Lecheb, A. Nour, S. Chellil, H. Mechakra, H. Kebir

Abstract:

In this paper, the experimental study for the instability of a separator rotor is presented, under dynamic loading response in the harmonic analysis condition. The analysis of the stress which operates the rotor is done. Calculations of different energies and the virtual work of the aerodynamic loads from the rotor are developed. Numerical calculations on the model develop of three dimensions prove that the defects effect has a negative effect on the stability of the rotor. Experimentally, the study of the rotor in the transient system allowed to determine the vibratory responses due to the unbalances and various excitations.

Keywords: rotor, frequency, finite element, specter

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2246 Laser - Ultrasonic Method for the Measurement of Residual Stresses in Metals

Authors: Alexander A. Karabutov, Natalia B. Podymova, Elena B. Cherepetskaya

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The theoretical analysis is carried out to get the relation between the ultrasonic wave velocity and the value of residual stresses. The laser-ultrasonic method is developed to evaluate the residual stresses and subsurface defects in metals. The method is based on the laser thermooptical excitation of longitudinal ultrasonic wave sand their detection by a broadband piezoelectric detector. A laser pulse with the time duration of 8 ns of the full width at half of maximum and with the energy of 300 µJ is absorbed in a thin layer of the special generator that is inclined relative to the object under study. The non-uniform heating of the generator causes the formation of a broadband powerful pulse of longitudinal ultrasonic waves. It is shown that the temporal profile of this pulse is the convolution of the temporal envelope of the laser pulse and the profile of the in-depth distribution of the heat sources. The ultrasonic waves reach the surface of the object through the prism that serves as an acoustic duct. At the interface ‚laser-ultrasonic transducer-object‘ the conversion of the most part of the longitudinal wave energy takes place into the shear, subsurface longitudinal and Rayleigh waves. They spread within the subsurface layer of the studied object and are detected by the piezoelectric detector. The electrical signal that corresponds to the detected acoustic signal is acquired by an analog-to-digital converter and when is mathematically processed and visualized with a personal computer. The distance between the generator and the piezodetector as well as the spread times of acoustic waves in the acoustic ducts are the characteristic parameters of the laser-ultrasonic transducer and are determined using the calibration samples. There lative precision of the measurement of the velocity of longitudinal ultrasonic waves is 0.05% that corresponds to approximately ±3 m/s for the steels of conventional quality. This precision allows one to determine the mechanical stress in the steel samples with the minimal detection threshold of approximately 22.7 MPa. The results are presented for the measured dependencies of the velocity of longitudinal ultrasonic waves in the samples on the values of the applied compression stress in the range of 20-100 MPa.

Keywords: laser-ultrasonic method, longitudinal ultrasonic waves, metals, residual stresses

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2245 Predicting College Students’ Happiness During COVID-19 Pandemic; Be optimistic and Well in College!

Authors: Michiko Iwasaki, Jane M. Endres, Julia Y. Richards, Andrew Futterman

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The present study aimed to examine college students’ happiness during COVID19-pandemic. Using the online survey data from 96 college students in the U.S., a regression analysis was conducted to predict college students’ happiness. The results indicated that a four-predictor model (optimism, college students’ subjective wellbeing, coronavirus stress, and spirituality) explained 57.9% of the variance in student’s subjective happiness, F(4,77)=26.428, p<.001, R2=.579, 95% CI [.41,.66]. The study suggests the importance of learned optimism among college students.

Keywords: COVID-19, optimism, spirituality, well-being

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2244 Etiology and Postnatal Management of Prenatal Hydronephrosis: A Study of Two Teaching Hospitals of Khyber Pakhtunkhwa

Authors: Saima Ali, Liaqat Ali, Nasir Orakzai

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Background: Hydronephrosis is the most common abnormal finding in the urinary tract on prenatal screening with Ultrasonography. The prenatal hydronephrosis is a diagnostic dilemma in differentiating between obstructive variant versus physiologic hydronephrosis. The assessment and prompt diagnosis of prenatal hydronephrosis is important because of the fact that untreated obstructive hydronephrosis usually leads to recurrent UTI, Urosepsis, deterioration of renal functions, non-functioning kidneys, and even end-stage renal disease. Objectives: To determine the etiology and outcome of postnatal treatment of children with prenatal hydronephrosis in two teaching hospitals of Khyber Pakhtunkhwa (KPK) Methods: It is a multicentric descriptive study that was conducted in department of Paediatrics in Kuwait teaching hospital Peshawar and Department of Urology in Institute of Kidney Diseases Hayatabad Medical Complex Peshawar from January 2008 till December 2010. Total numbers of 64 neonates were included in the study with the mean follow-up of 14.5 months. All the diagnostic data in prenatal, postnatal data, and operative and non-operative data were collected on structured Proforma and was analyzed on SPSS version 17. Results: Out of 64 patients, 39 (60.9 %) were male while 25 were female. 52 patients had unilateral while 12 patients had bilateral hydronephrosis. Based upon prenatal USG in term of AP diameter, 37 (57 %) patients had mild hydronephrosis (5-10 mm AP diameter), 14 patients had moderate hydronephrosis (10-15 mm AP diameter) while 13 patients had gross hydronephrosis (More than 15mm). Regarding etiology, 44(76 %) patients were labeled as physiologic hydronephrosis, 11 patients (9.3%) with PUJ obstruction, 5 patients with Vesicoureteric reflux (VUR) and 4 patients with posterior urethral valves. Surgery was performed in total of 15 (23.4%) patients that included open Pyeloplasty in 11 patients, Vesicostomy followed by posterior valve fulguration in 4 patients. All the patients of VUR treated medically. The severity in the grade of prenatal hydronephrosis is significantly associated with the need for definitive urological surgery p < 0.005. Ancillary procedures like percutaneous nephrostomy (PCN) were inserted 7 patients. Conclusions: Prenatal hydronephrosis is a common ailment associated with significant morbidity. Physiological Hydronephrosis and VUR can be successfully treated with medical treatment. However obstructive PUJ obstructions and posterior urethral valves require surgical correction with a good success rate.

Keywords: prenatal hydronephrosis, Pelviureteric Junction (PUJ) Obstruction, vesicoureteric reflux, posterior urethral valve, renography

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2243 Analytical Method Development and Validation of Stability Indicating Rp - Hplc Method for Detrmination of Atorvastatin and Methylcobalamine

Authors: Alkaben Patel

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The proposed RP-HPLC method is easy, rapid, economical, precise and accurate stability indicating RP-HPLC method for simultaneous estimation of Astorvastatin and Methylcobalamine in their combined dosage form has been developed.The separation was achieved by LC-20 AT C18(250mm*4.6mm*2.6mm)Colum and water (pH 3.5): methanol 70:30 as mobile phase, at a flow rate of 1ml/min. wavelength of this dosage form is 215nm.The drug is related to stress condition of hydrolysis, oxidation, photolysis and thermal degradation.

Keywords: RP- HPLC, atorvastatin, methylcobalamine, method, development, validation

Procedia PDF Downloads 336
2242 Experimental Study of Sand-Silt Mixtures with Torsional and Flexural Resonant Column Tests

Authors: Meghdad Payan, Kostas Senetakis, Arman Khoshghalb, Nasser Khalili

Abstract:

Dynamic properties of soils, especially at the range of very small strains, are of particular interest in geotechnical engineering practice for characterization of the behavior of geo-structures subjected to a variety of stress states. This study reports on the small-strain dynamic properties of sand-silt mixtures with particular emphasis on the effect of non-plastic fines content on the small strain shear modulus (Gmax), Young’s Modulus (Emax), material damping (Ds,min) and Poisson’s Ratio (v). Several clean sands with a wide range of grain size characteristics and particle shape are mixed with variable percentages of a silica non-plastic silt as fines content. Prepared specimens of sand-silt mixtures at different initial void ratios are subjected to sequential torsional and flexural resonant column tests with elastic dynamic properties measured along an isotropic stress path up to 800 kPa. It is shown that while at low percentages of fines content, there is a significant difference between the dynamic properties of the various samples due to the different characteristics of the sand portion of the mixtures, this variance diminishes as the fines content increases and the soil behavior becomes mainly silt-dominant, rendering no significant influence of sand properties on the elastic dynamic parameters. Indeed, beyond a specific portion of fines content, around 20% to 30% typically denoted as threshold fines content, silt is controlling the behavior of the mixture. Using the experimental results, new expressions for the prediction of small-strain dynamic properties of sand-silt mixtures are developed accounting for the percentage of silt and the characteristics of the sand portion. These expressions are general in nature and are capable of evaluating the elastic dynamic properties of sand-silt mixtures with any types of parent sand in the whole range of silt percentage. The inefficiency of skeleton void ratio concept in the estimation of small-strain stiffness of sand-silt mixtures is also illustrated.

Keywords: damping ratio, Poisson’s ratio, resonant column, sand-silt mixture, shear modulus, Young’s modulus

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2241 Genetic Divergence Study of Rice on the Basis of Various Morphological Traits

Authors: Muhammad Ashfaq, Muhammad Saleem Haider, Muhammad Ali, Muhammad Sajjad, Amna Ali, Urooj Mubashar

Abstract:

Phenotypic diversity was confirmed by measuring different morphological traits i.e. seed traits (seed length, seed width, seed thickness, seed length-width ratio, 1000 grain weight) and root-shoot traits (shoot length, root length, shoot fresh weight, root fresh weight, root-shoot ratio, root numbers and root thickness). Variance and association study of desirable traits determine the genotypic differences among the rice germplasm. All the traits showed significant differences among the genotypes. The traits were studied in Randomized complete block design (RCBD) at different water levels. Some traits showed positive correlation with each other and beneficial for increasing the yield and production of the crop. Seed thickness has positive correlation with seed length and seed width (r= 0.104**, r=0.246**). On the other hand, various root shoot traits showed positive highly significant association at different water levels i.e. root length, fresh root weight, root thickness, shoot thickness and root numbers. Our main focus to study the performance/correlation of root shoots traits under stress condition. Fresh root weight, shoot thickness and root numbers showed positive significant association with shoot length, root length, fresh root and shoot weight (r=0.2530**, r=0.2891**, r=0.4626**, r=0.4515**, r=0.5781**, r=0.7164**, r=0.0603**, r= 0.5570**, r=0.5824**). Long root length genotypes favors and suitable for drought stress conditions and screening of diverse genotypes for the further development of new plant material that performing well under different environmental conditions. After screening genetic diversity of potential rice, lines were studied to check the polymorphism by using some SSR markers. DNA was extracted, and PCR analyses were done to study PIC values and allelic diversity of the genotypes. The main objective of this study is to screen out the genotypes on the basis of various genotypic and phenotypic traits.

Keywords: rice, morphological traits, association, germplasm, genetic diversity, water levels, variation

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2240 The Effect of Online Learning During the COVID-19 Pandemic on Student Mental

Authors: Adelia Desi Agnesita

Abstract:

The advent of a new disease called covid-19 made many major changes in the world, one of which is the process of learning and teaching. Learning formerly offline but now is done online, which makes students need adaptation to the learning process. The covid-19 pandemic that occurs almost worldwide causes activities that involve many people to be avoided, one of which is learning to teach. In Indonesia, since March 2020, the process of college learning is turning into online/ long-distance learning. It's to prevent the spread of the covid-19. Student online learning presents some of the obstacles to poor signals, many of the tasks, lack of focus, difficulty sleeping, and resulting stress.

Keywords: learning, online, covid-19, pandemic

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2239 3D CFD Modelling of the Airflow and Heat Transfer in Cold Room Filled with Dates

Authors: Zina Ghiloufi, Tahar Khir

Abstract:

A transient three-dimensional computational fluid dynamics (CFD) model is developed to determine the velocity and temperature distribution in different positions cold room during pre-cooling of dates. The turbulence model used is the k-ω Shear Stress Transport (SST) with the standard wall function, the air. The numerical results obtained show that cooling rate is not uniform inside the room; the product at the medium of room has a slower cooling rate. This cooling heterogeneity has a large effect on the energy consumption during cold storage.

Keywords: CFD, cold room, cooling rate, dDates, numerical simulation, k-ω (SST)

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2238 Response Regimes and Vibration Mitigation in Equivalent Mechanical Model of Strongly Nonlinear Liquid Sloshing

Authors: Maor Farid, Oleg Gendelman

Abstract:

Equivalent mechanical model of liquid sloshing in partially-filled cylindrical vessel is treated in the cases of free oscillations and of horizontal base excitation. The model is designed to cover both the linear and essentially nonlinear sloshing regimes. The latter fluid behaviour might involve hydraulic impacts interacting with the inner walls of the tank. These impulsive interactions are often modeled by high-power potential and dissipation functions. For the sake of analytical description, we use the traditional approach by modeling the impacts with velocity-dependent restitution coefficient. This modelling is similar to vibro-impact nonlinear energy sink (VI NES) which was recently explored for its vibration mitigation performances and nonlinear response regimes. Steady-state periodic regimes and chaotic strongly modulated responses (CSMR) are detected. Those dynamical regimes were described by the system's slow motion on the slow invariant manifold (SIM). There is a good agreement between the analytical results and numerical simulations. Subsequently, Finite-Element (FE) method is used to determine and verify the model parameters and to identify dominant dynamical regimes, natural modes and frequencies. The tank failure modes are identified and critical locations are identified. Mathematical relation is found between degrees-of-freedom (DOFs) motion and the mechanical stress applied in the tank critical section. This is the prior attempt to take under consideration large-amplitude nonlinear sloshing and tank structure elasticity effects for design, regulation definition and resistance analysis purposes. Both linear (tuned mass damper, TMD) and nonlinear (nonlinear energy sink, NES) passive energy absorbers contribution to the overall system mitigation is firstly examined, in terms of both stress reduction and time for vibration decay.

Keywords: nonlinear energy sink (NES), reduced-order modelling, liquid sloshing, vibration mitigation, vibro-impact dynamics

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2237 Understanding Help Seeking among Black Women with Clinically Significant Posttraumatic Stress Symptoms

Authors: Glenda Wrenn, Juliet Muzere, Meldra Hall, Allyson Belton, Kisha Holden, Chanita Hughes-Halbert, Martha Kent, Bekh Bradley

Abstract:

Understanding the help seeking decision making process and experiences of health disparity populations with posttraumatic stress disorder (PTSD) is central to development of trauma-informed, culturally centered, and patient focused services. Yet, little is known about the decision making process among adult Black women who are non-treatment seekers as they are, by definition, not engaged in services. Methods: Audiotaped interviews were conducted with 30 African American adult women with clinically significant PTSD symptoms who were engaged in primary care, but not in treatment for PTSD despite symptom burden. A qualitative interview guide was used to elucidate key themes. Independent coding of themes mapped to theory and identification of emergent themes were conducted using qualitative methods. An existing quantitative dataset was analyzed to contextualize responses and provide a descriptive summary of the sample. Results: Emergent themes revealed that active mental avoidance, the intermittent nature of distress, ambivalence, and self-identified resilience as undermining to help seeking decisions. Participants were stuck within the help-seeking phase of ‘recognition’ of illness and retained a sense of “it is my decision” despite endorsing significant social and environmental negative influencers. Participants distinguished ‘help acceptance’ from ‘help seeking’ with greater willingness to accept help and importance placed on being of help to others. Conclusions: Elucidation of the decision-making process from the perspective of non-treatment seekers has implications for outreach and treatment within models of integrated and specialty systems care. The salience of responses to trauma symptoms and stagnation in the help seeking recognition phase are findings relevant to integrated care service design and community engagement.

Keywords: culture, help-seeking, integrated care, PTSD

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2236 Prevalence of ESBL E. coli Susceptibility to Oral Antibiotics in Outpatient Urine Culture: Multicentric, Analysis of Three Years Data (2019-2021)

Authors: Mazoun Nasser Rashid Al Kharusi, Nada Al Siyabi

Abstract:

Objectives: The main aim of this study is to Find the rate of susceptibility of ESBL E. coli causing UTI to oral antibiotics. Secondary objectives: Prevalence of ESBL E. coli from community urine samples, identify the best empirical oral antibiotics with the least resistance rate for UTI and identify alternative oral antibiotics for testing and utilization. Methods: This study is a retrospective descriptive study of the last three years in five major hospitals in Oman (Khowla Hospital, AN’Nahdha Hospital, Rustaq Hospital, Nizwa Hospital, and Ibri Hospital) equipped with a microbiologist. Inclusion criteria include all eligible outpatient urine culture isolates, excluding isolates from admitted patients with hospital-acquired urinary tract infections. Data was collected through the MOH database. The MOH hospitals are using different types of testing, automated methods like Vitek2 and manual methods. Vitek2 machine uses the principle of the fluorogenic method for organism identification and a turbidimetric method for susceptibility testing. The manual method is done by double disc diffusion for identifying ESBL and the disc diffusion method is for antibiotic susceptibility. All laboratories follow the clinical laboratory science institute (CLSI) guidelines. Analysis was done by SPSS statistical package. Results: Total urine cultures were (23048). E. coli grew in (11637) 49.6% of the urine, whereas (2199) 18.8% of those were confirmed as ESBL. As expected, the resistance rate to amoxicillin and cefuroxime is 100%. Moreover, the susceptibility of those ESBL-producing E. coli to nitrofurantoin, trimethoprim+sulfamethoxazole, ciprofloxacin and amoxicillin-clavulanate is progressing over the years; however, still low. ESBL E. coli was predominating in the female gender and those aged 66-74 years old throughout all the years. Other oral antibiotic options need to be explored and tested so that we add to the pool of oral antibiotics for ESBL E. coli causing UTI in the community. Conclusion: High rate of ESBL E. coli in urine from the community. The high resistance rates to oral antibiotics highlight the need for alternative treatment options for UTIs caused by these bacteria. Further research is needed to identify new and effective treatments for UTIs caused by ESBL-E. Coli.

Keywords: UTI, ESBL, oral antibiotics, E. coli, susceptibility

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2235 Hardness map of Human Tarsals, Meta Tarsals and Phalanges of Toes

Authors: Irfan Anjum Manarvi, Zahid Ali kaimkhani

Abstract:

Predicting location of the fracture in human bones has been a keen area of research for the past few decades. A variety of tests for hardness, deformation, and strain field measurement have been conducted in the past; but considered insufficient due to various limitations. Researchers, therefore, have proposed further studies due to inaccuracies in measurement methods, testing machines, and experimental errors. Advancement and availability of hardware, measuring instrumentation, and testing machines can now provide remedies to these limitations. The human foot is a critical part of the body exposed to various forces throughout its life. A number of products are developed for using it for protection and care, which many times do not provide sufficient protection and may itself become a source of stress due to non-consideration of the delicacy of bones in the feet. A continuous strain or overloading on feet may occur resulting to discomfort and even fracture. Mechanical properties of Tarsals, Metatarsals, and phalanges are, therefore, the primary area of consideration for all such design applications. Hardness is one of the mechanical properties which are considered very important to establish the mechanical resistance behavior of a material against applied loads. Past researchers have worked in the areas of investigating mechanical properties of these bones. However, their results were based on a limited number of experiments and taking average values of hardness due to either limitation of samples or testing instruments. Therefore, they proposed further studies in this area. The present research has been carried out to develop a hardness map of the human foot by measuring micro hardness at various locations of these bones. Results are compiled in the form of distance from a reference point on a bone and the hardness values for each surface. The number of test results is far more than previous studies and are spread over a typical bone to give a complete hardness map of these bones. These results could also be used to establish other properties such as stress and strain distribution in the bones. Also, industrial engineers could use it for design and development of various accessories for human feet health care and comfort and further research in the same areas.

Keywords: tarsals, metatarsals, phalanges, hardness testing, biomechanics of human foot

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2234 Genome Sequencing and Analysis of the Spontaneous Nanosilver Resistant Bacterium Proteus mirabilis Strain scdr1

Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

Abstract:

Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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2233 A Current Problem for Steel Bridges: Fatigue Assessment of Seams´ Repair

Authors: H. Pasternak, A. Chwastek

Abstract:

The paper describes the results from a research project about repair of welds. The repair was carried out by grinding the flawed seams and re-welding them. The main task was to determine the FAT classes of original state and after repair of seams according to the assessment procedures, such as nominal, structural and effective notch stress approach. The first part shows the results of the tests, the second part encloses numerical analysis and evaluation of results to determine the fatigue strength classes according to three assessment procedures.

Keywords: cyclic loading, fatigue crack, post-weld treatment, seams’ repair

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2232 Use of Pig as an Animal Model for Assessing the Differential MicroRNA Profiling in Kidney after Aristolochic Acid Intoxication

Authors: Daniela E. Marin, Cornelia Braicu, Gina C. Pistol, Roxana Cojocneanu-Petric, Ioana Berindan Neagoe, Mihail A. Gras, Ionelia Taranu

Abstract:

Aristolochic acid (AA) is a carcinogenic, mutagenic, and nephrotoxic compound commonly found in the Aristolochiaceae family of plants. AA is frequently associated with urothelial carcinoma of the upper urinary tract in human and animals and is considered as being responsible for Balkan Endemic Nephropathy. The pig provides a good animal model because the porcine urological system is very similar to that of humans, both in aspects of physiology and anatomy. MicroRNA (miRNA) are small non-coding RNAs that have an impact on a wide range of biological processes by regulating gene expression at post-transcriptional level. The objective of this study was to analyze the miRNA profiling in the kidneys of AA intoxicated swine. For this purpose, ten TOPIGS-40 crossbred weaned piglets, 4-week-old, male and females with an initial average body weight of 9.83 ± 0.5 kg were studied for 28 days. They were given ad libitum access to water and feed and randomly allotted to one of the following groups: control group (C) or aristolochic acid group (AA). They were fed a maize-soybean-meal-based diet contaminated or not with 0.25mgAA/kg. To profile miRNA in the kidneys of pigs, microarrays and bioinformatics approaches were applied to analyze the miRNA in the kidney of control and AA intoxicated pigs. After normalization, our results have shown that a total of 5 known miRNAs and 4 novel miRNAs had different profiling in the kidney of intoxicated animals versus control ones. Expression of miR-32-5p, miR-497-5p, miR-423-3p, miR-218-5p, miR-128-3p were up-regulated by 0.25mgAA/kg feed, while the expression of miR-9793-5p, miR-9835-3p, miR-9840-3p, miR-4334-5p was down-regulated. The microRNA profiling in kidney of intoxicated animals was associated with modified expression of target genes as: RICTOR, LASP1, SFRP2, DKK2, BMI1, RAF1, IGF1R, MAP2K1, WEE1, HDGF, BCL2, EIF4E etc, involved in cell division cycle, apoptosis, cell differentiation and cell migration, cell signaling, cancer etc. In conclusion, this study provides new data concerning the microRNA profiling in kidney after aristolochic acid intoxications with important implications for human and animal health.

Keywords: aristolochic acid, kidney, microRNA, swine

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2231 Resilient Design Solutions for Megathermal Climates of the Global South

Authors: Bobuchi Ken-Opurum

Abstract:

The impacts of climate change on urban settlements is growing. In the global south, communities are even more vulnerable and suffer there is an increased vulnerability from due to climate change disasters such as flooding and high temperatures. This is primarily due to high intensity rainfall, low-lying coasts, inadequate infrastructure, and limited resources. According to the Emergency Events Database, floods were the leading cause of disaster -based deaths in the global south between 2006 and 2015. This includes deaths from heat stress related health outcomes. Adapting to climate vulnerabilities is paramount in reducing the significant redevelopment costs from climate disasters. Governments and urban planners provide top-down approaches such as evacuation, and disaster and emergency communication. While they address infrastructure and public services, they are not always able to address the immediate and critical day to day needs of poor and vulnerable populations. There is growing evidence that some bottom-up strategies and grassroots initiatives of self-build housing such as in urban informal settlements are successful in coping and adapting to hydroclimatic impacts. However, these research findings are not consolidated and the evaluation of the resilience outcomes of the bottom-up strategies are limited. Using self-build housing as a model for sustainable and resilient urban planning, this research aimed to consolidate the flood and heat stress resilient design solutions, analyze the effectiveness of these solutions, and develop guidelines and methods for adopting these design solutions into mainstream housing in megathermal climates. The methodological approach comprised of analyses of over 40 ethnographic based peer reviewed literature, white papers, and reports between the years 2000 and 2019 to identify coping strategies and grassroots initiatives that have been applied by occupants and communities of the global south. The results of the research provide a consolidated source and prioritized list of the best bottom-up strategies for communities in megathermal climates to improve the lives of people in some of the most vulnerable places in the world.

Keywords: resilient, design, megathermal, climate change

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2230 Designing Next Generation Platforms for Recombinant Protein Production by Genome Engineering of Escherichia coli

Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

Abstract:

We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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