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Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies

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Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.

Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage

Procedia PDF Downloads 181

Authors: Hsueh Ping Peng

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End-stage renal disease most commonly occurs in the elderly population. Elderly people are approaching the end of their lives, and when facing major life-threatening situations, apart from aggressive medical treatment, they can also choose treatment methods such as hospice care to improve their quality of life. The purpose of this study was to investigate factors associated with the awareness and willingness to sign hospice and palliative care consent forms in elderly with end-stage renal disease. This study used both quantitative, cross-sectional study designs. In the quantitative section, 110 elderly patients (aged 65 or above) with end-stage renal disease receiving conventional hemodialysis were recruited as study participants from a medical center in Taipei City. Data were collected using structured questionnaires. Study tools included basic demographic data, questionnaires on the awareness and perception of hospice and palliative care, etc. After collecting the data, data analysis was conducted using SPSS 20.0 statistical software, including descriptive statistics, chi-square test, logistic regression, and other inferential statistics. The results showed that the average age of participants was 71.6 years old, more males than females, average years of dialysis was 6.1 years and most subjects rated their self-perceived health status as fair. Results of the study are summarized as follows: Elderly people with end-stage renal disease did not have sufficient knowledge and awareness about hospice and palliative care. Influencing factors included level of education, marital status, years of dialysis and age, etc. Demographic factors influencing the signing of consent forms included gender, marital status, and age, which all showed significant impacts. Factors taken into consideration when signing consent forms included awareness of hospice care, understanding the relevant definitions of hospice care, and understanding that consent may be modified or cancelled at any time; it was predicted that people who knew more about ways to receive hospice care or more related definitions were more willing to sign the consent forms. In the qualitative study section, 10 participants who signed the consent form, five male, and 5 female, between the ages of 65-90, have completed the semi-structured interviews. Analysis of the interviews revealed six themes: (1) passing away peacefully, (2) autonomy on arrangements of life and death, (3) unwillingness to increase family and social burden, (4) friends and relatives’ experience influencing the decision to give consent, (5) sharing information to facilitate the giving of consent, (6) facing each day with ease, to reflect the experience and factors of consideration for elderly with end-stage renal disease when signing consent forms. The results of this study provides the awareness, thoughts and feelings of elderly with end-stage renal disease on signing consent forms, and serve as a future reference for the dialysis unit to enhance the promotion of hospice and palliative care and related caregiving measures, thereby improving the quality of life and care for elderly people with end-stage renal disease.

Keywords: end-stage renal disease, hemodialysis, hospice and palliative care, awareness, willingness

Procedia PDF Downloads 171

Authors: Lucas Nascimento, Ana Rita, Margarida Soares, André Ribeiro, Zlatina Genisheva, Hugo Silva, Joana Carvalho

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The footwear industry is an essential fashion industry, focusing on producing various types of footwear, such as shoes, boots, sandals, sneakers, and slippers. Global footwear consumption has doubled every 20 years since the 1950s. It is estimated that in 1950, each person consumed one new pair of shoes yearly; by 2005, over 20 billion pairs of shoes were consumed. To meet global footwear demand, production reached $24.2 billion, equivalent to about $74 per person in the United States. This means three new pairs of shoes per person worldwide. The issue of footwear waste is related to the fact that shoe production can generate a large amount of waste, much of which is difficult to recycle or reuse. This waste includes scraps of leather, fabric, rubber, plastics, toxic chemicals, and other materials. The search for alternative solutions for waste treatment and valorization is increasingly relevant in the current context, mainly when focused on utilizing waste as a source of substitute materials. From the perspective of the new circular economy paradigm, this approach is of utmost importance as it aims to preserve natural resources and minimize the environmental impact associated with sending waste to landfills. In this sense, the incorporation of waste into industrial sectors that allow for the recovery of large volumes, such as road construction, becomes an urgent and necessary solution from an environmental standpoint. This study explores the use of plastic waste from the footwear industry as a substitute for virgin polymers in bitumen modification, a solution that presents a more sustainable future. Replacing conventional polymers with plastic waste in asphalt composition reduces the amount of waste sent to landfills and offers an opportunity to extend the lifespan of road infrastructures. By incorporating waste into construction materials, reducing the consumption of natural resources and the emission of pollutants is possible, promoting a more circular and efficient economy. In the initial phase of this study, waste materials from end-of-life footwear were selected, and plastic waste with the highest potential for application was separated. Based on a literature review, EVA (ethylene vinyl acetate) and PU (polyurethane) were identified as the polymers suitable for modifying 50/70 classification bitumen. Each polymer was analysed at concentrations of 3% and 5%. The production process involved the polymer's fragmentation to a size of 4 millimetres after heating the materials to 180 ºC and mixing for 10 minutes at low speed. After was mixed for 30 minutes in a high-speed mixer. The tests included penetration, softening point, viscosity, and rheological assessments. With the results obtained from the tests, the mixtures with EVA demonstrated better results than those with PU, as EVA had more resistance to temperature, a better viscosity curve and a greater elastic recovery in rheology.

Keywords: footwear waste, hot asphalt pavement, modified bitumen, polymers

Procedia PDF Downloads 22

Authors: Timur S. Zaripov, Artur K. Gilfanov, Sergei S. Sazhin, Steven M. Begg, Morgan R. Heikal

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The calculation of the concentration of inertial droplets in complex flows is encountered in the modelling of numerous engineering and environmental phenomena; for example, fuel droplets in internal combustion engines and airborne pollutant particles. The results of recent research, focused on the development of methods for calculating concentration and their implementation in the commercial CFD code, ANSYS Fluent, is presented here. The study is motivated by the investigation of the mixture preparation processes in internal combustion engines with direct injection of fuel sprays. Two methods are used in our analysis; the Fully Lagrangian method (also known as the Osiptsov method) and the Eulerian approach. The Osiptsov method predicts droplet concentrations along path lines by solving the equations for the components of the Jacobian of the Eulerian-Lagrangian transformation. This method significantly decreases the computational requirements as it does not require counting of large numbers of tracked droplets as in the case of the conventional Lagrangian approach. In the Eulerian approach the average droplet velocity is expressed as a function of the carrier phase velocity as an expansion over the droplet response time and transport equation can be solved in the Eulerian form. The advantage of the method is that droplet velocity can be found without solving additional partial differential equations for the droplet velocity field. The predictions from the two approaches were compared in the analysis of the problem of a dilute gas-droplet flow around an infinitely long, circular cylinder. The concentrations of inertial droplets, with Stokes numbers of 0.05, 0.1, 0.2, in steady-state and transient laminar flow conditions, were determined at various Reynolds numbers. In the steady-state case, flows with Reynolds numbers of 1, 10, and 100 were investigated. It has been shown that the results predicted using both methods are almost identical at small Reynolds and Stokes numbers. For larger values of these numbers (Stokes — 0.1, 0.2; Reynolds — 10, 100) the Eulerian approach predicted a wider spread in concentration in the perturbations caused by the cylinder that can be attributed to the averaged droplet velocity field. The transient droplet flow case was investigated for a Reynolds number of 200. Both methods predicted a high droplet concentration in the zones of high strain rate and low concentrations in zones of high vorticity. The maxima of droplet concentration predicted by the Osiptsov method was up to two orders of magnitude greater than that predicted by the Eulerian method; a significant variation for an approach widely used in engineering applications. Based on the results of these comparisons, the Osiptsov method has resulted in a more precise description of the local properties of the inertial droplet flow. The method has been applied to the analysis of the results of experimental observations of a liquid gasoline spray at representative fuel injection pressure conditions. The preliminary results show good qualitative agreement between the predictions of the model and experimental data.

Keywords: internal combustion engines, Eulerian approach, fully Lagrangian approach, gasoline fuel sprays, droplets and particle concentrations

Procedia PDF Downloads 262

Authors: A. Matiushkina, A. Bazhenova, I. Litvinov, E. Kornilova, A. Dubavik, A. Orlova

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Magnetic-luminescent complexes based on superparamagnetic iron oxide nanoparticles (SPIONs) and semiconductor quantum dots (QDs) have been recognized as a new class of materials that have high potential in modern medicine. These materials can serve for theranostics of oncological diseases, and also as a target agent for drug delivery. They combine the qualities characteristic of magnetic nanoparticles, that is, magneto-controllability and the ability to local heating under the influence of an external magnetic field, as well as phosphors, due to luminescence of which, for example, early tumor imaging is possible. The complexity of creating complexes is the energy transfer between particles, which quenches the luminescence of QDs in complexes with SPIONs. In this regard, a relatively new type of alloyed (CdₓZn₁₋ₓSeᵧS₁₋ᵧ)-ZnS QDs is used in our work. The presence of a sufficiently thick gradient semiconductor shell in alloyed QDs makes it possible to reduce the probability of energy transfer from QDs to SPIONs in complexes. At the same time, Forster Resonance Energy Transfer (FRET) is a perfect instrument to confirm the formation of complexes based on QDs and different-type energy acceptors. The formation of complexes in the aprotic bipolar solvent dimethyl sulfoxide is ensured by the coordination of the carboxyl group of the stabilizing QD molecule (L-cysteine) on the surface iron atoms of the SPIONs. An analysis of the photoluminescence (PL) spectra has shown that a sequential increase in the SPIONs concentration in the samples is accompanied by effective quenching of the luminescence of QDs. However, it has not confirmed the formation of complexes yet, because of a decrease in the PL intensity of QDs due to reabsorption of light by SPIONs. Therefore, a study of the PL kinetics of QDs at different SPIONs concentrations was made, which demonstrates that an increase in the SPIONs concentration is accompanied by a symbatic reduction in all characteristic PL decay times. It confirms the FRET from QDs to SPIONs, which indicates the QDs/SPIONs complex formation, rather than a spontaneous aggregation of QDs, which is usually accompanied by a sharp increase in the percentage of the QD fraction with the shortest characteristic PL decay time. The complexes have been studied by the magnetic circular dichroism (MCD) spectroscopy that allows one to estimate the response of magnetic material to the applied magnetic field and also can be useful to check SPIONs aggregation. An analysis of the MCD spectra has shown that the complexes have zero residual magnetization, which is an important factor for using in biomedical applications, and don't contain SPIONs aggregates. Cell penetration, biocompatibility, and stability of QDs/SPIONs complexes in cancer cells have been studied using HeLa cell line. We have found that the complexes penetrate in HeLa cell and don't demonstrate cytotoxic effect up to 25 nM concentration. Our results clearly demonstrate that alloyed (CdₓZn₁₋ₓSeᵧS₁₋ᵧ)-ZnS QDs can be successfully used in complexes with SPIONs reached new hybrid nanostructures, which combine bright luminescence for tumor imaging and magnetic properties for targeted drug delivery and magnetic hyperthermia of tumors. Acknowledgements: This work was supported by the Ministry of Science and Higher Education of Russian Federation, goszadanie no. 2019-1080 and was financially supported by Government of Russian Federation, Grant 08-08.

Keywords: alloyed quantum dots, magnetic circular dichroism, magneto-luminescent complexes, superparamagnetic iron oxide nanoparticles

Procedia PDF Downloads 122

Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

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Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

Procedia PDF Downloads 335

Authors: Suparna Roy, Anatharaman Perumal

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The industrial effluent with high amount of heavy metals is known to have adverse effects on the environment. For the removal of heavy metals from aqueous environment, different conventional treatment technologies had been applied gradually which are not economically beneficial and also produce huge quantity of toxic chemical sludge. So, bio-sorption of heavy metals by marine plant is an eco-friendly innovative and alternative technology for removal of these pollutants from aqueous environment. The aim of this study is to evaluate the capacity of heavy metals accumulation and removal by some selected marine macroalgae (seaweeds) from marine environment. Methods: Seaweeds Acanthophora spicifera (Vahl.) Boergesen, Codium tomentosum Stackhouse, Halimeda gracilis Harvey ex. J. Agardh, Gracilaria opuntia Durairatnam.nom. inval. Valoniopsis pachynema (Martens) Boergesen, Caulerpa racemosa var. macrophysa (Sonder ex Kutzing) W. R. Taylor and Hydroclathrus clathratus (C. Agardh) Howe were collected from Olaikuda (09°17.526'N-079°19.662'E), Rameshwaram, south east coast of India during post monsoon period (April’2016). Seaweeds were washed with sterilized and filtered in-situ seawater repeatedly to remove all the epiphytes and debris and clean seaweeds were kept for shade drying for one week. The dried seaweeds were grinded to powder, and one gm powder seaweeds were taken in a 250ml conical flask, and 8 ml of 10 % HNO3 (70 % pure) was added to each sample and kept in room temperature (28 ̊C) for 24 hours and then samples were heated in hotplate at 120 ̊C, boiled to evaporate up to dryness and 20 ml of Nitric acid: Percholoric acid in 4:1 were added to it and again heated to hotplate at 90 ̊C up to evaporate to dryness, then samples were kept in room temperature for few minutes to cool and 10ml 10 % HNO3 were added to it and kept for 24 hours in cool and dark place and filtered with Whatman (589/2) filter paper and the filtrates were collected in 250ml clean conical flask and diluted accurately to 25 ml volume with double deionised water and triplicate of each sample were analysed with Inductively-Coupled plasma analysis (ICP-OES) to analyse total eleven heavy metals (Ag, Cd, B, Cu, Mn, Co, Ni, Cr, Pb, Zn, and Al content of the specified species and data were statistically evaluated for standard deviation. Results: Acanthophora spicifera contains highest amount of Ag (0.1± 0.2 mg/mg) followed by Cu (0.16±0.01 mg/mg), Mn (1.86±0.02 mg/mg), B (3.59±0.2 mg/mg), Halimeda gracilis showed highest accumulation of Al (384.75±0.12mg/mg), Valoniopsis pachynema accumulates maximum amount of Co (0.12±0.01 mg/mg), Zn (0.64±0.02 mg/mg), Caulerpa racemosa var. macrophysa contains Zn (0.63±0.01), Cr (0.26±0.01 mg/mg ), Ni (0.21±0.05), Pb (0.16±0.03 ) and Cd ( 0.02±00 ). Hydroclathrus clathratus, Codium tomentosum and Gracilaria opuntia also contain adequate amount of heavy metals. Conclusions: The mentioned species of seaweeds are contributing important role for decreasing the heavy metals pollution in marine environment by bioaccumulation. So, we can utilise this species to remove excess amount of heavy metals from polluted area.

Keywords: heavy metals pollution, seaweeds, bioaccumulation, eco-friendly, phyco-remediation

Procedia PDF Downloads 236

Authors: Iryna Atamaniuk, Hannah Boysen, Nils Wieczorek, Natalia Politaeva, Iuliia Bazarnova, Kerstin Kuchta

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Over the last decade due to climate change and a strategy of natural resources preservation, the interest for the aquatic biomass has dramatically increased. Along with mitigation of the environmental pressure and connection of waste streams (including CO₂ and heat emissions), microalgae bioeconomy can supply food, feed, as well as the pharmaceutical and power industry with number of value-added products. Furthermore, in comparison to conventional biomass, microalgae can be cultivated in wide range of conditions without compromising food and feed production, thus addressing issues associated with negative social and the environmental impacts. This paper presents the state-of-the art technology for microalgae bioeconomy from cultivation process to production of valuable components and by-streams. Microalgae Chlorella sorokiniana were cultivated in the pilot-scale innovation concept in Hamburg (Germany) using different systems such as race way pond (5000 L) and flat panel reactors (8 x 180 L). In order to achieve the optimum growth conditions along with suitable cellular composition for the further extraction of the value-added components, process parameters such as light intensity, temperature and pH are continuously being monitored. On the other hand, metabolic needs in nutrients were provided by addition of micro- and macro-nutrients into a medium to ensure autotrophic growth conditions of microalgae. The cultivation was further followed by downstream process and extraction of lipids, proteins and saccharides. Lipids extraction is conducted in repeated-batch semi-automatic mode using hot extraction method according to Randall. As solvents hexane and ethanol are used at different ratio of 9:1 and 1:9, respectively. Depending on cell disruption method along with solvents ratio, the total lipids content showed significant variations between 8.1% and 13.9 %. The highest percentage of extracted biomass was reached with a sample pretreated with microwave digestion using 90% of hexane and 10% of ethanol as solvents. Proteins content in microalgae was determined by two different methods, namely: Total Kejadahl Nitrogen (TKN), which further was converted to protein content, as well as Bradford method using Brilliant Blue G-250 dye. Obtained results, showed a good correlation between both methods with protein content being in the range of 39.8–47.1%. Characterization of neutral and acid saccharides from microalgae was conducted by phenol-sulfuric acid method at two wavelengths of 480 nm and 490 nm. The average concentration of neutral and acid saccharides under the optimal cultivation conditions was 19.5% and 26.1%, respectively. Subsequently, biomass residues are used as substrate for anaerobic digestion on the laboratory-scale. The methane concentration, which was measured on the daily bases, showed some variations for different samples after extraction steps but was in the range between 48% and 55%. CO₂ which is formed during the fermentation process and after the combustion in the Combined Heat and Power unit can potentially be used within the cultivation process as a carbon source for the photoautotrophic synthesis of biomass.

Keywords: bioeconomy, lipids, microalgae, proteins, saccharides

Procedia PDF Downloads 249

Authors: Kuo-Kai Lin, Po-Lun Chang

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Research Background and Purpose: Following the development of the Internet and multimedia, the Internet and information technology have become crucial avenues of modern communication and knowledge acquisition. The advantages of using mobile devices for learning include making learning borderless and accessible. Mobile learning has become a trend in disease management and health promotion in recent years. End-stage renal disease (ESRD) is an irreversible chronic disease, and patients who do not receive kidney transplants can only rely on hemodialysis or peritoneal dialysis to survive. Due to the complexities in caregiving for patients with ESRD that stem from their advanced age and other comorbidities, the patients’ incapacity of self-care leads to an increase in the need to rely on their families or primary caregivers, although whether the primary caregivers adequately understand and implement patient care is a topic of concern. Therefore, this study explored whether primary caregivers’ health care provisions can be improved through the intervention of an automatic speech intelligent system, thereby improving the objective health outcomes of patients undergoing long-term dialysis. Method: This study developed an automatic speech intelligent system with healthcare functions such as health information voice prompt, two-way feedback, real-time push notification, and health information delivery. Convenience sampling was adopted to recruit eligible patients from a hemodialysis center at a regional teaching hospital as research participants. A one-group pretest-posttest design was adopted. Descriptive and inferential statistics were calculated from the demographic information collected from questionnaires answered by patients and primary caregivers, and from a medical record review, a health care scale (recorded six months before and after the implementation of intervention measures), a subjective health assessment, and a report of objective physiological indicators. The changes in health care behaviors, subjective health status, and physiological indicators before and after the intervention of the proposed automatic speech intelligent system were then compared. Conclusion and Discussion: The preliminary automatic speech intelligent system developed in this study was tested with 20 pretest patients at the recruitment location, and their health care capacity scores improved from 59.1 to 72.8; comparisons through a nonparametric test indicated a significant difference (p < .01). The average score for their subjective health assessment rose from 2.8 to 3.3. A survey of their objective physiological indicators discovered that the compliance rate for the blood potassium level was the most significant indicator; its average compliance rate increased from 81% to 94%. The results demonstrated that this automatic speech intelligent system yielded a higher efficacy for chronic disease care than did conventional health education delivered by nurses. Therefore, future efforts will continue to increase the number of recruited patients and to refine the intelligent system. Future improvements to the intelligent system can be expected to enhance its effectiveness even further.

Keywords: automatic speech intelligent system for health care, primary caregiver, long-term hemodialysis, health care capabilities, health outcomes

Procedia PDF Downloads 114

Authors: Galatee Levadoux, Trevor Benson, Chris Worrall

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With the rise of public awareness on climate change comes an increasing demand for renewable sources of energy. As a result, the wind power sector is striving to manufacture longer, more efficient and reliable wind turbine blades. Currently, one of the leading causes of blade failure in service is improper cure of the resin during manufacture. The infusion process creating the main part of the composite blade structure remains a critical step that is yet to be monitored in real time. This stage consists of a viscous resin being drawn into a mould under vacuum, then undergoing a curing reaction until solidification. Successful infusion assumes the resin fills all the voids and cures completely. Given that the electrical properties of the resin change significantly during its solidification, both the filling of the mould and the curing reaction are susceptible to be followed using dieletrometry. However, industrially available dielectrics sensors are currently too small to monitor the entire surface of a wind turbine blade. The aim of the present research project is to scale up the dielectric sensor technology and develop a device able to monitor the manufacturing process of large composite structures, assessing the conformity of the blade before it even comes out of the mould. An array of flat copper wires acting as electrodes are embedded in a polymer matrix fixed in an infusion mould. A multi-frequency analysis from 1 Hz to 10 kHz is performed during the filling of the mould with an epoxy resin and the hardening of the said resin. By following the variations of the complex admittance Y*, the filling of the mould and curing process are monitored. Results are compared to numerical simulations of the sensor in order to validate a virtual cure-monitoring system. The results obtained by drawing glycerol on top of the copper sensor displayed a linear relation between the wetted length of the sensor and the complex admittance measured. Drawing epoxy resin on top of the sensor and letting it cure at room temperature for 24 hours has provided characteristic curves obtained when conventional interdigitated sensor are used to follow the same reaction. The response from the developed sensor has shown the different stages of the polymerization of the resin, validating the geometry of the prototype. The model created and analysed using COMSOL has shown that the dielectric cure process can be simulated, so long as a sufficient time and temperature dependent material properties can be determined. The model can be used to help design larger sensors suitable for use with full-sized blades. The preliminary results obtained with the sensor prototype indicate that the infusion and curing process of an epoxy resin can be followed with the chosen configuration on a scale of several decimeters. Further work is to be devoted to studying the influence of the sensor geometry and the infusion parameters on the results obtained. Ultimately, the aim is to develop a larger scale sensor able to monitor the flow and cure of large composite panels industrially.

Keywords: composite manufacture, dieletrometry, epoxy, resin infusion, wind turbine blades

Procedia PDF Downloads 169

Authors: Márcia Afonso, Cristina Fael, Marisa Dinis-Almeida

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Population growth in cities has led to an increase in the infrastructures construction, including buildings and roadways. This aspect leads directly to the soils waterproofing. In turn, changes in precipitation patterns are developing into higher and more frequent intensities. Thus, these two conjugated aspects decrease the rainwater infiltration into soils and increase the volume of surface runoff. The practice of green and sustainable urban solutions has encouraged research in these areas. The porous asphalt pavement, as a green infrastructure, is part of practical solutions set to address urban challenges related to land use and adaptation to climate change. In this field, permeable pavements with porous asphalt mixtures (PA) have several advantages in terms of reducing the runoff generated by the floods. The porous structure of these pavements, compared to a conventional asphalt pavement, allows the rainwater infiltration in the subsoil, and consequently, the water quality improvement. This green infrastructure solution can be applied in cities, particularly in streets or parking lots to mitigate the floods effects. Over the years, the pores of these pavements can be filled by sediment, reducing their function in the rainwater infiltration. Thus, double layer porous asphalt (DLPA) was developed to mitigate the clogging effect and facilitate the water infiltration into the lower layers. This study intends to deepen the knowledge of the performance of DLPA when subjected to clogging. The experimental methodology consisted on four evaluation phases of the DLPA infiltration capacity submitted to three precipitation events (100, 200 and 300 mm/h) in each phase. The evaluation first phase determined the behavior after DLPA construction. In phases two and three, two 500 g/m² clogging cycles were performed, totaling a 1000 g/m² final simulation. Sand with gradation accented in fine particles was used as clogging material. In the last phase, the DLPA was subjected to simple sweeping and vacuuming maintenance. A precipitation simulator, type sprinkler, capable of simulating the real precipitation was developed for this purpose. The main conclusions show that the DLPA has the capacity to drain the water, even after two clogging cycles. The infiltration results of flows lead to an efficient performance of the DPLA in the surface runoff attenuation, since this was not observed in any of the evaluation phases, even at intensities of 200 and 300 mm/h, simulating intense precipitation events. The infiltration capacity under clogging conditions decreased about 7% on average in the three intensities relative to the initial performance that is after construction. However, this was restored when subjected to simple maintenance, recovering the DLPA hydraulic functionality. In summary, the study proved the efficacy of using a DLPA when it retains thicker surface sediments and limits the fine sediments entry to the remaining layers. At the same time, it is guaranteed the rainwater infiltration and the surface runoff reduction and is therefore a viable solution to put into practice in permeable pavements.

Keywords: clogging, double layer porous asphalt, infiltration capacity, rainfall intensity

Procedia PDF Downloads 495

Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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Authors: Ola Hall, Ibrahim Wahab, Thorsteinn Rognvaldsson, Mattias Ohlsson

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The subfield of poverty and welfare estimation that applies machine learning tools and methods on satellite imagery is a nascent but rapidly growing one. This is in part driven by the sustainable development goal, whose overarching principle is that no region is left behind. Among other things, this requires that welfare levels can be accurately and rapidly estimated at different spatial scales and resolutions. Conventional tools of household surveys and interviews do not suffice in this regard. While they are useful for gaining a longitudinal understanding of the welfare levels of populations, they do not offer adequate spatial coverage for the accuracy that is needed, nor are their implementation sufficiently swift to gain an accurate insight into people and places. It is this void that satellite imagery fills. Previously, this was near-impossible to implement due to the sheer volume of data that needed processing. Recent advances in machine learning, especially the deep learning subtype, such as deep neural networks, have made this a rapidly growing area of scholarship. Despite their unprecedented levels of performance, such models lack transparency and explainability and thus have seen limited downstream applications as humans generally are apprehensive of techniques that are not inherently interpretable and trustworthy. While several studies have demonstrated the superhuman performance of AI models, none has directly compared the performance of such models and human readers in the domain of poverty studies. In the present study, we directly compare the performance of human readers and a DL model using different resolutions of satellite imagery to estimate the welfare levels of demographic and health survey clusters in Tanzania, using the wealth quintile ratings from the same survey as the ground truth data. The cluster-level imagery covers all 608 cluster locations, of which 428 were classified as rural. The imagery for the human readers was sourced from the Google Maps Platform at an ultra-high resolution of 0.6m per pixel at zoom level 18, while that of the machine learning model was sourced from the comparatively lower resolution Sentinel-2 10m per pixel data for the same cluster locations. Rank correlation coefficients of between 0.31 and 0.32 achieved by the human readers were much lower when compared to those attained by the machine learning model – 0.69-0.79. This superhuman performance by the model is even more significant given that it was trained on the relatively lower 10-meter resolution satellite data while the human readers estimated welfare levels from the higher 0.6m spatial resolution data from which key markers of poverty and slums – roofing and road quality – are discernible. It is important to note, however, that the human readers did not receive any training before ratings, and had this been done, their performance might have improved. The stellar performance of the model also comes with the inevitable shortfall relating to limited transparency and explainability. The findings have significant implications for attaining the objective of the current frontier of deep learning models in this domain of scholarship – eXplainable Artificial Intelligence through a collaborative rather than a comparative framework.

Keywords: poverty prediction, satellite imagery, human readers, machine learning, Tanzania

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Authors: Inna Kornienko, Svetlana Guryeva, Anatoly Shekhter, Elena Petersen

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Transplantation is an effective treatment option for patients suffering from different end-stage diseases. However, it is plagued by a constant shortage of donor organs and the subsequent need of a lifelong immunosuppressive therapy for the patient. Currently some researchers look towards using of pig organs to replace human organs for transplantation since the matrix derived from porcine organs is a convenient substitute for the human matrix. As an initial step to create a new ex vivo tissue engineered model, optimized protocols have been created to obtain organ-specific acellular matrices and evaluated their potential as tissue engineered scaffolds for culture of normal cells and tumor cell lines. These protocols include decellularization by perfusion in a bioreactor system and immersion-agitation on an orbital shaker with use of various detergents (SDS, Triton X-100) and freezing. Complete decellularization – in terms of residual DNA amount – is an important predictor of probability of immune rejection of materials of natural origin. However, the signs of cellular material may still remain within the matrix even after harsh decellularization protocols. In this regard, the matrices obtained from tissues of low-immunogenic pigs with α3Galactosyl-tranferase gene knock out (GalT-KO) may be a promising alternative to native animal sources. The research included a study of induced effect of frozen and fresh fragments of GalT-KO skin on healing of full-thickness plane wounds in 80 rats. Commercially available wound dressings (Ksenoderm, Hyamatrix and Alloderm) as well as allogenic skin were used as a positive control and untreated wounds were analyzed as a negative control. The results were evaluated on the 4th day after grafting, which corresponds to the time of start of normal wound epithelization. It has been shown that a non-specific immune response in models treated with GalT-Ko pig skin was milder than in all the control groups. Research has been performed to measure technical skin characteristics: stiffness and elasticity properties, corneometry, tevametry, and cutometry. These metrics enabled the evaluation of hydratation level, corneous layer husking level, as well as skin elasticity and micro- and macro-landscape. These preliminary data may contribute to development of personalized transplantable organs from GalT-Ko pigs with significantly limited potential of immune rejection. By applying growth factors to a decellularized skin sample it is possible to achieve various regenerative effects based on the particular situation. In this particular research BMP2 and Heparin-binding EGF-like growth factor have been used. Ideally, a bioengineered organ must be biocompatible, non-immunogenic and support cell growth. Porcine organs are attractive for xenotransplantation if severe immunologic concerns can be bypassed. The results indicate that genetically modified pig tissues with knock-outed α3Galactosyl-tranferase gene may be used for production of low-immunogenic matrix suitable for transplantation.

Keywords: decellularization, low-immunogenic, matrix, scaffolds, transplants

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Authors: Rosemary R. Carlton, Roxane Caron

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Almost three decades ago, bell hooks (1994) identified the classroom as “the most radical space of possibility in the academy”. A feminist scholar, educator, and activist, hooks urged educators to transgress the boundaries of what might be customary or considered acceptable in teaching, thus encouraging the pursuit of new ways of knowing and different strategies for sharing knowledge. This paper reflects upon a particular response to hooks’ still relevant call for transgression in teaching. Specifically, this paper reports on the design, implementation, and preliminary analysis of a social work course integrating active pedagogy and transnational exchange to encourage students’ critical thinking and autonomous learning in their development as social workers in a global context. The bachelor’s level course, Pratiques spécifiques: Projet international, was developed collaboratively across three francophone institutions of higher learning in Belgium, Canada, and France: the Haute École de Namur-Liège-Luxembourg (Hénallux); the Université de Montréal; and, the Institut d’enseignement supérieur et professionnel, l’IRTS Paris Île-de-France. The driving aims of the course are to promote autonomous learning and critical thinking through a lens of transnational understandings of social problems -competencies indispensable to students’ development as social workers. The course is offered to two paired cohorts, one addressing the subject of “migrations” (Canada/France) and the other the subject of “sexual exploitation” (Canada/Belgium). Through the adaptation of a critical pedagogy of problem-based learning, students are called upon to actively engage in acquiring and applying knowledge to respond to “real life” social issues relating to migration or sexual exploitation. At the conclusion of the course, each cohort of students is brought together for a week-long intensive period of transnational exchange either at the Université de Montréal in Canada or at Hénallux in Belgium. Extending the bounds of the classroom across international borders allows students novel opportunities to deepen and expand their understandings of issues relating to predefined social issues and to critically examine associated social work practices. The paper opens with a presentation of the social work course. Specifically, the authors will outline their adaptation of a pedagogy of problem-based learning integrating transnational exchange in the design and implementation of the course. Returning to hooks’ notion of transgression in teaching, the paper offers a preliminary analysis of how and with what effect the course provides opportunities to transgress hierarchical student-teacher relationships; transgress conventional modes of learning to explore diverse sources of knowledge and transgress the walls of the university to engage with and learn from local and global partners. The paper concludes with a consideration of the potential influence of such transgressions in teaching for students’ development of critical thinking in their practice of social work in global context.

Keywords: active learning, critical pedagogy, social work intervention, transnational learning

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Authors: Bernice Jeanne Lottering, Yi-Wen Lin

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Introduction: Chronic pain has a definitive lack of objective parameters in the measurement and treatment efficacy of diseases such as Fibromyalgia (FM). Persistent widespread pain and generalized tenderness are the characteristic symptoms affecting a large majority of the global population, particularly females. This disease has indicated a refractory tendency to conventional treatment ventures, largely resultant from a lack of etiological and pathogenic understanding of the disease development. Emerging evidence indicates that the central nervous system (CNS) plays a critical role in the amplification of pain signals and the neurotransmitters associated therewith. Various stimuli have been found to activate the channels existent on nociceptor terminals, thereby actuating nociceptive impulses along the pain pathways. The transient receptor potential vanalloid 1 (TRPV1) channel functions as a molecular integrator for numerous sensory inputs, such as nociception, and was explored in the current study. Current intervention approaches face a multitude challenges, ranging from effective therapeutic interventions to the limitation of pathognomonic criteria resultant from incomplete understanding and partial evidence on the mechanisms of action of FM. It remains unclear whether electroacupuncture (EA) plays an integral role in the functioning of the TRPV1 pathway, and whether or not it can reduce the chronic pain induced by FM. Aims: The aim of this study was to explore the mechanisms underlying the activation and modulation of the TRPV1 channel pathway in a cold stress model of FM applied to a murine model. Furthermore, the effect of EA in the treatment of mechanical and thermal pain, as expressed in FM was also to be investigated. Methods: 18 C57BL/6 wild type and 6 TRPV1 knockout (KO) mice, aged 8-12 weeks, were exposed to an intermittent cold stress-induced fibromyalgia-like pain model, with or without EA treatment at ZusanLi ST36 (2Hz/20min) on day 3 to 5. Von Frey and Hargreaves behaviour tests were implemented in order to analyze the mechanical and thermal pain thresholds on day 0, 3 and 5 in control group (C), FM group (FM), FM mice with EA treated group (FM + EA) and FM in KO group. Results: An increase in mechanical and thermal hyperalgesia was observed in the FM, EA and KO groups when compared to the control group. This initial increase was reduced in the EA group, which directs focus at the treatment efficacy of EA in nociceptive sensitization, and the analgesic effect EA has attenuating FM associated pain. Discussion: An increase in the nociceptive sensitization was observed through higher withdrawal thresholds in the von Frey mechanical test and the Hargreaves thermal test. TRPV1 function in mice has been scientifically associated with these nociceptive conduits, and the increased behaviour test results suggest that TRPV1 upregulation is central to the FM induced hyperalgesia. This data was supported by the decrease in sensitivity observed in results of the TRPV1 KO group. Moreover, the treatment of EA showed a decrease in this FM induced nociceptive sensitization, suggesting TRPV1 upregulation and overexpression can be attenuated by EA at bilateral ST36. This evidence compellingly implies that the analgesic effect of EA is associated with TRPV1 downregulation.

Keywords: fibromyalgia, electroacupuncture, TRPV1, nociception

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Authors: Chui-Hsin Chen, Yu-Ting Chen

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Special concentrically braced frames is one of the seismic load resisting systems, which dissipates seismic energy when bracing members within the frames undergo yielding and buckling while sustaining their axial tension and compression load capacities. Most of the inelastic deformation of a buckling bracing member concentrates in the mid-length region. While experiencing cyclic loading, the region dissipates most of the seismic energy being input into the frame. Such a concentration makes the braces vulnerable to failure modes associated with low-cycle fatigue. In this research, a strategy to improve the cyclic behavior of the conventional steel bracing member is proposed by filling the Hollow Structural Shape (HSS) member with reinforcement. It prevents the local section from concentrating large plastic deformation caused by cyclic loading. The infill helps spread over the plastic hinge region into a wider area hence postpone the initiation of local buckling or even the rupture of the braces. The finite element method is introduced to simulate the complicated bracing member behavior and member-versus-infill interaction under cyclic loading. Fifteen 3-D-element-based models are built by ABAQUS software. The verification of the FEM model is done with unreinforced (UR) HSS bracing members’ cyclic test data and aluminum honeycomb plates’ bending test data. Numerical models include UR and filled HSS bracing members with various compactness ratios based on the specification of AISC-2016 and AISC-1989. The primary variables to be investigated include the relative bending stiffness and the material of the filling reinforcement. The distributions of von Mises stress and equivalent plastic strain (PEEQ) are used as indices to tell the strengths and shortcomings of each model. The result indicates that the change of relative bending stiffness of the infill is much more influential than the change of material in use to increase the energy dissipation capacity. Strengthen the relative bending stiffness of the reinforcement results in additional energy dissipation capacity to the extent of 24% and 46% in model based on AISC-2016 (16-series) and AISC-1989 (89-series), respectively. HSS members with infill show growth in 𝜂Local Buckling, normalized energy cumulated until the happening of local buckling, comparing to UR bracing members. The 89-series infill-reinforced members have more energy dissipation capacity than unreinforced 16-series members by 117% to 166%. The flexural rigidity of infills should be less than 29% and 13% of the member section itself for 16-series and 89-series bracing members accordingly, thereby guaranteeing the spread over of the plastic hinge and the happening of it within the reinforced section. If the parameters are properly configured, the ductility, energy dissipation capacity, and fatigue-life of HSS SCBF bracing members can be improved prominently by the infill-reinforced method.

Keywords: special concentrically braced frames, HSS, cyclic loading, infill reinforcement, finite element analysis, PEEQ

Procedia PDF Downloads 96

Authors: Mahla Azimi

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Introduction: Post-bladder catheter infection is a common and significant healthcare-associated infection that affects individuals with indwelling urinary catheters. These infections can lead to various complications, including urinary tract infections (UTIs), bacteremia, sepsis, and increased morbidity and mortality rates. This article aims to provide a comprehensive review of post-bladder catheter infections, including their causes, risk factors, clinical presentation, diagnosis, treatment options, and preventive measures. Causes and Risk Factors: Post-bladder catheter infections primarily occur due to the colonization of microorganisms on the surface of the urinary catheter. The most common pathogens involved are Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus species. Several risk factors contribute to the development of these infections, such as prolonged catheterization duration, improper insertion technique, poor hygiene practices during catheter care, compromised immune system function in patients with underlying conditions or immunosuppressive therapy. Clinical Presentation: Patients with post-bladder catheter infections may present with symptoms such as fever, chills, malaise, suprapubic pain or tenderness, and cloudy or foul-smelling urine. In severe cases or when left untreated for an extended period of time, patients may develop more severe symptoms like hematuria or signs of systemic infection. Diagnosis: The diagnosis of post-bladder catheter infection involves a combination of clinical evaluation and laboratory investigations. Urinalysis is crucial in identifying pyuria (presence of white blood cells) and bacteriuria (presence of bacteria). A urine culture is performed to identify the causative organism(s) and determine its antibiotic susceptibility profile. Treatment Options: Prompt initiation of appropriate antibiotic therapy is essential in managing post-bladder catheter infections. Empirical treatment should cover common pathogens until culture results are available. The choice of antibiotics should be guided by local antibiogram data to ensure optimal therapy. In some cases, catheter removal may be necessary, especially if the infection is recurrent or associated with severe complications. Preventive Measures: Prevention plays a vital role in reducing the incidence of post-bladder catheter infections. Strategies include proper hand hygiene, aseptic technique during catheter insertion and care, regular catheter maintenance, and timely removal of unnecessary catheters. Healthcare professionals should also promote patient education regarding self-care practices and signs of infection. Conclusion: Post-bladder catheter infections are a significant healthcare concern that can lead to severe complications and increased healthcare costs. Early recognition, appropriate diagnosis, and prompt treatment are crucial in managing these infections effectively. Implementing preventive measures can significantly reduce the incidence of post-bladder catheter infections and improve patient outcomes. Further research is needed to explore novel strategies for prevention and management in this field.

Keywords: post-bladder catheter infection, urinary tract infection, bacteriuria, indwelling urinary catheters, prevention

Procedia PDF Downloads 85

Authors: Matej Komanec, Tomas Nemecek, Dmytro Suslov, Petr Chvojka, Stanislav Zvanovec

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Nonlinear phenomena in the near- and mid-infrared region are attracting scientific attention mainly due to the supercontinuum generation possibilities and subsequent utilizations for ultra-wideband applications like e.g. absorption spectroscopy or optical coherence tomography. Thulium-based fiber lasers provide access to high-power ultrashort pump pulses in the vicinity of 2000 nm, which can be easily exploited for various nonlinear applications. The paper presents a simulation and experimental study of a pulsed thulium laser based for near-infrared (NIR) and mid-infrared (MIR) nonlinear applications in specialty optical fibers. In the first part of the paper the thulium laser is discussed. The thulium laser is based on a gain-switched seed-laser and a series of amplification stages for obtaining output peak powers in the order of kilowatts for pulses shorter than 200 ps in full-width at half-maximum. The pulsed thulium laser is first studied in a simulation software, focusing on seed-laser properties. Afterward, a pre-amplification thulium-based stage is discussed, with the focus of low-noise signal amplification, high signal gain and eliminating pulse distortions during pulse propagation in the gain medium. Following the pre-amplification stage a second gain stage is evaluated with incorporating a thulium-fiber of shorter length with increased rare-earth dopant ratio. Last a power-booster stage is analyzed, where the peak power of kilowatts should be achieved. Examples of analytical study are further validated by the experimental campaign. The simulation model is further corrected based on real components – parameters such as real insertion-losses, cross-talks, polarization dependencies, etc. are included. The second part of the paper evaluates the utilization of nonlinear phenomena, their specific features at the vicinity of 2000 nm, compared to e.g. 1550 nm, and presents supercontinuum modelling, based on the thulium laser pulsed output. Supercontinuum generation simulation is performed and provides reasonably accurate results, once fiber dispersion profile is precisely defined and fiber nonlinearity is known, furthermore input pulse shape and peak power must be known, which is assured thanks to the experimental measurement of the studied thulium pulsed laser. The supercontinuum simulation model is put in relation to designed and characterized specialty optical fibers, which are discussed in the third part of the paper. The focus is placed on silica and mainly on non-silica fibers (fluoride, chalcogenide, lead-silicate) in their conventional, microstructured or tapered variants. Parameters such as dispersion profile and nonlinearity of exploited fibers were characterized either with an accurate model, developed in COMSOL software or by direct experimental measurement to achieve even higher precision. The paper then combines all three studied topics and presents a possible application of such a thulium pulsed laser system working with specialty optical fibers.

Keywords: nonlinear phenomena, specialty optical fibers, supercontinuum generation, thulium laser

Procedia PDF Downloads 324

Authors: V. Giacometti, R. Mirabelli, V. Patera, D. Pinci, A. Sarti, A. Sciubba, G. Traini, M. Marafini

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The particle therapy (PT) is a very modern technique of non invasive radiotherapy mainly devoted to the treatment of tumours untreatable with surgery or conventional radiotherapy, because localised closely to organ at risk (OaR). Nowadays, PT is available in about 55 centres in the word and only the 20\% of them are able to treat with carbon ion beam. However, the efficiency of the ion-beam treatments is so impressive that many new centres are in construction. The interest in this powerful technology lies to the main characteristic of PT: the high irradiation precision and conformity of the dose released to the tumour with the simultaneous preservation of the adjacent healthy tissue. However, the beam interactions with the patient produce a large component of secondary particles whose additional dose has to be taken into account during the definition of the treatment planning. Despite, the largest fraction of the dose is released to the tumour volume, a non-negligible amount is deposed in other body regions, mainly due to the scattering and nuclear interactions of the neutrons within the patient body. One of the main concerns in PT treatments is the possible occurrence of secondary malignant neoplasm (SMN). While SMNs can be developed up to decades after the treatments, their incidence impacts directly life quality of the cancer survivors, in particular in pediatric patients. Dedicated Treatment Planning Systems (TPS) are used to predict the normal tissue toxicity including the risk of late complications induced by the additional dose released by secondary neutrons. However, no precise measurement of secondary neutrons flux is available, as well as their energy and angular distributions: an accurate characterization is needed in order to improve TPS and reduce safety margins. The project MONDO (MOnitor for Neutron Dose in hadrOntherapy) is devoted to the construction of a secondary neutron tracker tailored to the characterization of that secondary neutron component. The detector, based on the tracking of the recoil protons produced in double-elastic scattering interactions, is a matrix of thin scintillating fibres, arranged in layer x-y oriented. The final size of the object is 10 x 10 x 20 cm3 (squared 250µm scint. fibres, double cladding). The readout of the fibres is carried out with a dedicated SPAD Array Sensor (SBAM) realised in CMOS technology by FBK (Fondazione Bruno Kessler). The detector is under development as well as the SBAM sensor and it is expected to be fully constructed for the end of the year. MONDO will make data tacking campaigns at the TIFPA Proton Therapy Center of Trento, at the CNAO (Pavia) and at HIT (Heidelberg) with carbon ion in order to characterize the neutron component and predict the additional dose delivered on the patients with much more precision and to drastically reduce the actual safety margins. Preliminary measurements with charged particles beams and MonteCarlo FLUKA simulation will be presented.

Keywords: secondary neutrons, particle therapy, tracking detector, elastic scattering

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Authors: Huey-Yeu Yan, Ching-Ying Lee, Hung-Ru Lin

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Background: Physical assessment is a vital clinical nursing competence. The gap between conventional teaching method and the way e-generation students’ preferred could be bridged owing to the support of Internet technology, i.e. interacting with online media to manage learning works. Nursing instructors in the wake of new learning pattern of the e-generation students are challenged to actively adjust and make teaching contents and methods more versatile. Objective: The objective of this research is to explore the effects on teaching and learning with live webcast-assisted on a specific topic, Physical Assessment technique, on a designated group of young nursing majors. It’s hoped that, with a way of nursing instructing, more versatile learning resources may be provided to facilitate self-directed learning. Design: This research adopts a cross-sectional descriptive survey. The instructor demonstrated physical assessment techniques and operation procedures via live webcast broadcasted online to all students. It increased both the off-time interaction between teacher and students concerning teaching materials. Methods: A convenient sampling was used to recruit a total of 52 nursing-majors at a certain university. The nursing majors took two-hour classes of Physical Assessment per week for 18 weeks (36 hrs. in total). The instruction covered four units with live webcasting and then conducted an online anonymous survey of learning outcomes by questionnaire. The research instrument was the online questionnaire, covering three major domains—online media used, learning outcome evaluation and evaluation result. The data analysis was conducted via IBM SPSS Statistics Version 2.0. The descriptive statistics was undertaken to describe the analysis of basic data and learning outcomes. Statistical methods such as descriptive statistics, t-test, ANOVA, and Pearson’s correlation were employed in verification. Results: Results indicated the following five major findings. (1) learning motivation, about four fifth of the participants agreed the online instruction resources are very helpful in improving learning motivation and raising the learning interest. (2) learning needs, about four fifth of participants agreed it was helpful to plan self-directed practice after the instruction, and meet their needs of repetitive learning and/or practice at their leisure time. (3) learning effectiveness, about two third agreed it was helpful to reduce pre-exam anxiety, and improve their test scores. (4) course objects, about three fourth agreed that it was helpful to achieve the goal of ‘executing the complete Physical Assessment procedures with proper skills’. (5) finally, learning reflection, about all of participants agreed this experience of online instructing, learning, and practicing is beneficial to them, they recommend instructor to share with other nursing majors, and they will recommend it to fellow students too. Conclusions: Live webcasting is a low-cost, convenient, efficient and interactive resource to facilitate nursing majors’ motivation of learning, need of self-directed learning and practice, outcome of learning. When live webcasting is integrated into nursing teaching, it provides an opportunity of self-directed learning to promote learning effectiveness, as such to fulfill the teaching objective.

Keywords: innovative teaching, learning effectiveness, live webcasting, physical assessment technique

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Authors: Sthanu Mahadev, Wen Chan, Melanie Lim

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Anisotropy dominated continuous-fiber composite materials have garnered attention in numerous mechanical and aerospace structural applications. Tailored mechanical properties in advanced composites can exhibit superiority in terms of stiffness-to-weight ratio, strength-to-weight ratio, low-density characteristics, coupled with significant improvements in fatigue resistance as opposed to metal structure counterparts. Extensive research has demonstrated their core potential as more than just mere lightweight substitutes to conventional materials. Prior work done by Mahadev and Chan focused on formulating a modified composite shell theory based prognosis methodology for investigating the structural response of thin-walled circular cylindrical shell type composite configurations under in-plane mechanical loads respectively. The prime motivation to develop this theory stemmed from its capability to generate simple yet accurate closed-form analytical results that can efficiently characterize circular composite shell construction. It showcased the development of a novel mathematical framework to analytically identify the location of the centroid for thin-walled, open cross-section, curved composite shells that were characterized by circumferential arc angle, thickness-to-mean radius ratio, and total laminate thickness. Ply stress variations for curved cylindrical shells were analytically examined under the application of centric tensile and bending loading. This work presents a cost-effective, small-platform experimental methodology by taking advantage of the full-field measurement capability of digital image correlation (DIC) for an accurate assessment of key mechanical parameters such as in-plane mechanical stresses and strains, centroid location etc. Mechanical property measurement of advanced composite materials can become challenging due to their anisotropy and complex failure mechanisms. Full-field displacement measurements are well suited for characterizing the mechanical properties of composite materials because of the complexity of their deformation. This work encompasses the fabrication of a set of curved cylindrical shell coupons, the design and development of a novel test-fixture design and an innovative experimental methodology that demonstrates the capability to very accurately predict the location of centroid in such curved composite cylindrical strips via employing a DIC based strain measurement technique. Error percentage difference between experimental centroid measurements and previously estimated analytical centroid results are observed to be in good agreement. The developed analytical modified-shell theory provides the capability to understand the fundamental behavior of thin-walled cylindrical shells and offers the potential to generate novel avenues to understand the physics of such structures at a laminate level.

Keywords: anisotropy, composites, curved cylindrical shells, digital image correlation

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Authors: Recep Kesli, Cengiz Demir, Onur Turkyilmaz, Hayriye Tokay

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Objective: Gram-negative rods are a large group of bacteria, and include many families, genera, and species. Most clinical isolates belong to the family Enterobacteriaceae. Resistance due to the production of extended-spectrum β-lactamases (ESBLs) is a difficulty in the handling of Enterobacteriaceae infections, but other mechanisms of resistance are also emerging, leading to multidrug resistance and threatening to create panresistant species. We aimed in this study to evaluate resistance rates of Gram-negative rods bacteria isolated from clinical specimens in Microbiology Laboratory, Afyon Kocatepe University, ANS Research and Practice Hospital, between October 2012 and September 2015. Methods: The Gram-negative rods strains were identified by conventional methods and VITEK 2 automated identification system (bio-Mérieux, Marcy l’etoile, France). Antibiotic resistance tests were performed by both the Kirby-Bauer disk-diffusion and automated Antimicrobial Susceptibility Testing (AST, bio-Mérieux, Marcy l’etoile, France) methods. Disk diffusion results were evaluated according to the standards of Clinical and Laboratory Standards Institute (CLSI). Results: Of the totally isolated 1.701 Enterobacteriaceae strains 1434 (84,3%) were Klebsiella pneumoniae, 171 (10%) were Enterobacter spp., 96 (5.6%) were Proteus spp., and 639 Nonfermenting gram negatives, 477 (74.6%) were identified as Pseudomonas aeruginosa, 135 (21.1%) were Acinetobacter baumannii and 27 (4.3%) were Stenotrophomonas maltophilia. The ESBL positivity rate of the totally studied Enterobacteriaceae group were 30.4%. Antibiotic resistance rates for Klebsiella pneumoniae were as follows: amikacin 30.4%, gentamicin 40.1%, ampicillin-sulbactam 64.5%, cefepime 56.7%, cefoxitin 35.3%, ceftazidime 66.8%, ciprofloxacin 65.2%, ertapenem 22.8%, imipenem 20.5%, meropenem 20.5 %, and trimethoprim-sulfamethoxazole 50.1%, and for 114 Enterobacter spp were detected as; amikacin 26.3%, gentamicin 31.5%, cefepime 26.3%, ceftazidime 61.4%, ciprofloxacin 8.7%, ertapenem 8.7%, imipenem 12.2%, meropenem 12.2%, and trimethoprim-sulfamethoxazole 19.2 %. Resistance rates for Proteus spp. were: 24,3% meropenem, 26.2% imipenem, 20.2% amikacin 10.5% cefepim, 33.3% ciprofloxacin and levofloxacine, 31.6% ceftazidime, 20% ceftriaxone, 15.2% gentamicin, 26.6% amoxicillin-clavulanate, and 26.2% trimethoprim-sulfamethoxale. Resistance rates of P. aeruginosa was found as follows: Amikacin 32%, gentamicin 42 %, imipenem 43%, merpenem 43%, ciprofloxacin 50%, levofloxacin 52%, cefepim 38%, ceftazidim 63%, piperacillin/tacobactam 85%, for Acinetobacter baumannii; Amikacin 53.3%, gentamicin 56.6 %, imipenem 83%, merpenem 86%, ciprofloxacin 100%, ceftazidim 100%, piperacillin/tacobactam 85 %, colisitn 0 %, and for S. malthophilia; levofloxacin 66.6 % and trimethoprim/sulfamethoxozole 0 %. Conclusions: This study showed that resistance in Gram-negative rods was a serious clinical problem in our hospital and suggested the need to perform typification of the isolated bacteria with susceptibility testing regularly in the routine laboratory procedures. This application guided to empirical antibiotic treatment choices truly, as a consequence of the reality that each hospital shows different resistance profiles.

Keywords: antibiotic resistance, gram negative rods, ESBL, VITEK 2

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Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

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Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

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Authors: Joanna Marie A. Paulino-Morente, Vaneza Valentina L. Penolio, Grace Sabado

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Ovarian cancer is extremely hard to diagnose in its early stages, and those afflicted at the time of diagnosis are typically asymptomatic and in the late stages of the disease, with metastasis to other organs. Ovarian cancers often occur sporadically, with only 5% associated with hereditary mutations. Mutations in the BRCA1 and BRCA2 tumor suppressor genes have been found to be responsible for the majority of hereditary ovarian cancers. One type of ovarian tumor is Malignant Mixed Mullerian Tumor (MMMT), which is a very rare and aggressive type, accounting for only 1% of all ovarian cancers. Reported is a case of a 43-year-old G3P3 (3003), who came into our institution due to a 2-month history of difficulty of breathing. Family history reveals that her eldest and younger sisters both died of ovarian malignancy, with her younger sister having a histopathology report of endometrioid ovarian carcinoma, left ovary stage IIIb. She still has 2 asymptomatic sisters. Physical examination pointed to pleural effusion of right lung, and presence of bilateral ovarian new growth, which had a Sassone score of 13. Admitting Diagnosis was G3P3 (3003), Ovarian New Growth, bilateral, Malignant; Pleural effusion secondary to malignancy. BRCA was requested to establish a hereditary mutation; however, the patient had no funds. Once the patient was stabilized, TAHBSO with surgical staging was performed. Intraoperatively, the pelvic cavity was occupied by firm, irregularly shaped ovaries, with a colorectal metastasis. Microscopic sections from both ovaries and the colorectal metastasis had pleomorphic tumor cells lined by cuboidal to columnar epithelium exhibiting glandular complexity, displaying nuclear atypia and increased nuclear-cytoplasmic ratio, which are infiltrating the stroma, consistent with the features of Malignant Mixed Mullerian Tumor, since MMMT is composed histologically of malignant epithelial and sarcomatous elements. In conclusion, discussed is the clinic-pathological feature of a patient with primary ovarian Malignant Mixed Mullerian Tumor, a rare malignancy comprising only 1% of all ovarian neoplasms. Also, by understanding the hereditary ovarian cancer syndromes and its relation to this patient, it cannot be overemphasized that a comprehensive family history is really fundamental for early diagnosis. The familial association of the disease, given that the patient has two sisters who were diagnosed with an advanced stage of ovarian cancer and succumbed to the disease at a much earlier age than what is reported in the general population, points to a possible hereditary syndrome which occurs in only 5% of ovarian neoplasms. In a low-resource setting, being in a third world country, the following will be recommended for monitoring and/or screening women who are at high risk for developing ovarian cancer, such as the remaining sisters of the patient: 1) Physical examination focusing on the breast, abdomen, and rectal area every 6 months. 2) Transvaginal sonography every 6 months. 3) Mammography annually. 4) CA125 for postmenopausal women. 5) Genetic testing for BRCA1 and BRCA2 will be reserved for those who are financially capable.

Keywords: BRCA, hereditary breast-ovarian cancer syndrome, malignant mixed mullerian tumor, ovarian cancer

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Authors: Fabienne Samyn, Pauline Tranchard, Sophie Duquesne, Emilie Goncalves, Bruno Estebe, Serge Boubigot

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Stringent fire safety regulations are enforced in the aeronautical industry due to the consequences that potential fire event on an aircraft might imply. This is so much true that the fire issue is considered right from the design of the aircraft structure. Due to the incorporation of an increasing amount of polymer matrix composites in replacement of more conventional materials like metals, the nature of the fire risks is changing. The choice of materials used is consequently of prime importance as well as the evaluation of its resistance to fire. The fire testing is mostly done using the so-called certification tests according to standards such as the ISO2685:1998(E). The latter describes a protocol to evaluate the fire resistance of structures located in fire zone (ability to withstand fire for 5min). The test consists in exposing an at least 300x300mm² sample to an 1100°C propane flame with a calibrated heat flux of 116kW/m². This type of test is time-consuming, expensive and gives access to limited information in terms of fire behavior of the materials (pass or fail test). Consequently, it can barely be used for material development purposes. In this context, the laboratory UMET in collaboration with industrial partners has developed a horizontal and a vertical small-scale instrumented fire benches for the characterization of the fire behavior of composites. The benches using smaller samples (no more than 150x150mm²) enables to cut downs costs and hence to increase sampling throughput. However, the main added value of our benches is the instrumentation used to collect useful information to understand the behavior of the materials. Indeed, measurements of the sample backside temperature are performed using IR camera in both configurations. In addition, for the vertical set up, a complete characterization of the degradation process, can be achieved via mass loss measurements and quantification of the gasses released during the tests. These benches have been used to characterize and study the fire behavior of aeronautical carbon/epoxy composites. The horizontal set up has been used in particular to study the performances and durability of protective intumescent coating on 2mm thick 2D laminates. The efficiency of this approach has been validated, and the optimized coating thickness has been determined as well as the performances after aging. Reductions of the performances after aging were attributed to the migration of some of the coating additives. The vertical set up has enabled to investigate the degradation process of composites under fire. An isotropic and a unidirectional 4mm thick laminates have been characterized using the bench and post-fire analyses. The mass loss measurements and the gas phase analyses of both composites do not present significant differences unlike the temperature profiles in the thickness of the samples. The differences have been attributed to differences of thermal conductivity as well as delamination that is much more pronounced for the isotropic composite (observed on the IR-images). This has been confirmed by X-ray microtomography. The developed benches have proven to be valuable tools to develop fire safe composites.

Keywords: aeronautical carbon/epoxy composite, durability, intumescent coating, small-scale ‘ISO 2685 like’ fire resistance test, X-ray microtomography

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Authors: Maksim Barsakov

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Introduction. In modern plastic surgery, there are a large number of breast lift techniques.Due to the spreading information about the "side effects" of silicone implants, interest in implant-free mastopexy is increasing year after year. However, despite the variety of techniques, patients sometimes do not get full satisfaction from the results of mastopexy because of the unexpressed filling of the upper pole, extended anchoring postoperative scars and sometimes because of obtaining an aesthetically unattractive breast shape. The stability of the result after mastopexy depends on many factors, including postoperative rehabilitation. Stability of weight and hormonal background, stretchability of tissues. The high recurrence rate of ptosis and short-term aesthetic effect of mastopexy indicate the urgency of improving surgical techniques and increasing the stabilization of breast tissue. Purpose of the study. To develop and introduce into practice a technique of mastopexy based on the use of a modified Ribeiro flap, as well as elements of tissue movement and fixation designed to increase the stability of postoperative mastopexy. In addition, to give indications for the application of this surgical technique. Materials and Methods. it operated on 103 patients aged 18 to 53 years from 2019 to 2023 according to the reported method. These were patients with primary mastopexy, secondary mastopexy, and also patient with implant removal and one-stage mastopexy. The patients were followed up for 12 months to assess the stability of the result. Results and their discussion. Observing the patients, we noted greater stability of the breast shape and upper pole filling compared to the conventional classical methods. We did not have to resort to anchoring scars. In 90 percent of cases, a inverted T-shape scar was used. In 10 percent, the J-scar was used. The quantitative distribution of complications identified among the operated patients is as follows: worsened healing of the junction of vertical and horizontal sutures at the period of 1-1.5 months after surgery - 15 patients; at treatment with ointment method healing was observed in 7-30 days; permanent loss of NAC sensitivity - 0 patients; vascular disorders in the area of NAC/areola necrosis - 0 patients; marginal necrosis of the areola-2 patients. independent healing within 3-4 weeks without aesthetic defects. Aesthetically unacceptable mature scars-3 patients; partial liponecrosis of the autoflap unilaterally - 1 patient. recurrence of ptosis - 1 patient (after weight loss of 12 kg). In the late postoperative period, 2 patients became pregnant, gave birth, and no lactation problems were observed. Conclusion. Thus, in the world of plastic surgery methods of breast lift continue to improve, which is especially relevant in modern times, due to the increased attention to this operation. The author's proposed method of mastopexy with glandular autoflap allows obtaining in most cases a stable result, a fuller breast shape, avoiding the presence of extended anchoring scars, and also preserves the possibility of lactation. The author of this article has obtained a patent for invention for this method of mastopexy.

Keywords: mastopexy, mammoplasty, autoflap, personal technique

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Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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Authors: Yi-Hao Wong, Li-Li Chan, Chee-Onn Leong, Stephen Ambu, Joon-Wah Mak, Priyasashi Sahu

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Background: Acanthamoeba is a genus of amoebae which lives as a free-living in nature or as a human pathogen that causes severe brain and eye infections. Virulence potential of Acanthamoeba is not constant and can change with growth conditions. DNA methylation, an epigenetic process which adds methyl groups to DNA, is used by eukaryotic cells, including several human parasites to control their gene expression. We used qPCR, siRNA gene silencing, and RNA sequencing (RNA-Seq) to study DNA-methyltransferase gene family (DNMT) in order to indicate the possibility of its involvement in programming Acanthamoeba virulence potential. Methods: A virulence-attenuated Acanthamoeba isolate (designation: ATCC; original isolate: ATCC 50492) was subjected to mouse passages to restore its pathogenicity; a virulence-reactivated isolate (designation: AC/5) was generated. Several established factors associated with Acanthamoeba virulence phenotype were examined to confirm the succession of reactivation process. Differential gene expression of DNMT between ATCC and AC/5 isolates was performed by qPCR. Silencing on DNMT gene expression in AC/5 isolate was achieved by siRNA duplex. Total RNAs extracted from ATCC, AC/5, and siRNA-treated (designation: si-146) were subjected to RNA-Seq for comparative transcriptomic analysis in order to identify the genome-wide effect of DNMT in regulating Acanthamoeba gene expression. qPCR was performed to validate the RNA-Seq results. Results: Physiological and cytophatic assays demonstrated an increased in virulence potential of AC/5 isolate after mouse passages. DNMT gene expression was significantly higher in AC/5 compared to ATCC isolate (p ≤ 0.01) by qPCR. si-146 duplex reduced DNMT gene expression in AC/5 isolate by 30%. Comparative transcriptome analysis identified the differentially expressed genes, with 3768 genes in AC/5 vs ATCC isolate; 2102 genes in si-146 vs AC/5 isolate and 3422 genes in si-146 vs ATCC isolate, respectively (fold-change of ≥ 2 or ≤ 0.5, p-value adjusted (padj) < 0.05). Of these, 840 and 1262 genes were upregulated and downregulated, respectively, in si-146 vs AC/5 isolate. Eukaryotic orthologous group (KOG) assignments revealed a higher percentage of downregulated gene expression in si-146 compared to AC/5 isolate, were related to posttranslational modification, signal transduction and energy production. Gene Ontology (GO) terms for those downregulated genes shown were associated with transport activity, oxidation-reduction process, and metabolic process. Among these downregulated genes were putative genes encoded for heat shock proteins, transporters, ubiquitin-related proteins, proteins for vesicular trafficking (small GTPases), and oxidoreductases. Functional analysis of similar predicted proteins had been described in other parasitic protozoa for their survival and pathogenicity. Decreased expression of these genes in si146-treated isolate may account in part for Acanthamoeba reduced pathogenicity. qPCR on 6 selected genes upregulated in AC/5 compared to ATCC isolate corroborated the RNA sequencing findings, indicating a good concordance between these two analyses. Conclusion: To the best of our knowledge, this study represents the first genome-wide analysis of DNA methylation and its effects on gene expression in Acanthamoeba spp. The present data indicate that DNA methylation has substantial effect on global gene expression, allowing further dissection of the genome-wide effects of DNA-methyltransferase gene in regulating Acanthamoeba pathogenicity.

Keywords: Acanthamoeba, DNA methylation, RNA sequencing, virulence

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Authors: Jothilakshmy Nagammal

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This research paper deals with the different case studies, where the Form-Based Codes are adopted in general and the different implementation methods in particular are discussed to develop a method for formulating a new planning model. The organizing principle of the Form-Based Codes, the transect is used to zone the city into various context specific transects. An approach is adopted to develop the new planning model, city Specific Planning Model (CSPM), as a tool to achieve sustainability for any city in general. A case study comparison method in terms of the planning tools used, the code process adopted and the various control regulations implemented in thirty two different cities are done. The analysis shows that there are a variety of ways to implement form-based zoning concepts: Specific plans, a parallel or optional form-based code, transect-based code /smart code, required form-based standards or design guidelines. The case studies describe the positive and negative results from based zoning, Where it is implemented. From the different case studies on the method of the FBC, it is understood that the scale for formulating the Form-Based Code varies from parts of the city to the whole city. The regulating plan is prepared with the organizing principle as the transect in most of the cases. The various implementation methods adopted in these case studies for the formulation of Form-Based Codes are special districts like the Transit Oriented Development (TOD), traditional Neighbourhood Development (TND), specific plan and Street based. The implementation methods vary from mandatory, integrated and floating. To attain sustainability the research takes the approach of developing a regulating plan, using the transect as the organizing principle for the entire area of the city in general in formulating the Form-Based Codes for the selected Special Districts in the study area in specific, street based. Planning is most powerful when it is embedded in the broader context of systemic change and improvement. Systemic is best thought of as holistic, contextualized and stake holder-owned, While systematic can be thought of more as linear, generalisable, and typically top-down or expert driven. The systemic approach is a process that is based on the system theory and system design principles, which are too often ill understood by the general population and policy makers. The system theory embraces the importance of a global perspective, multiple components, interdependencies and interconnections in any system. In addition, the recognition that a change in one part of a system necessarily alters the rest of the system is a cornerstone of the system theory. The proposed regulating plan taking the transect as an organizing principle and Form-Based Codes to achieve sustainability of the city has to be a hybrid code, which is to be integrated within the existing system - A Systemic Approach with a Systematic Process. This approach of introducing a few form based zones into a conventional code could be effective in the phased replacement of an existing code. It could also be an effective way of responding to the near-term pressure of physical change in “sensitive” areas of the community. With this approach and method the new Context Specific Planning Model is created towards achieving sustainability is explained in detail this research paper.

Keywords: context based planning model, form based code, transect, systemic approach

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