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Commenced in January 2007
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Paper Count: 2859

Search results for: live and dead bacteria

129 Ruminal Fermentation of Biologically Active Nitrate- and Nitro-Containing Forages

Authors: Robin Anderson, David Nisbet

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Nitrate, 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) are biologically active chemicals that can accumulate naturally in rangeland grasses forages consumed by grazing cattle, sheep and goats. While toxic to livestock if accumulations and amounts consumed are high enough, particularly in animals having no recent exposure to the forages, these chemicals are known to be potent inhibitors of methane-producing bacteria inhabiting the rumen. Consequently, there is interest in examining their potential use as anti-methanogenic compounds to decrease methane emissions by grazing ruminants. Presently, rumen microbes, collected freshly from a cannulated Holstein cow maintained on 50:50 corn based concentrate:alfalfa diet were mixed (10 mL fluid) in 18 x 150 mm crimp top tubes with 0.5 of high nitrate-containing barley (Hordeum vulgare; containing 272 µmol nitrate per g forage dry matter), and NPA- or NPOH- containing milkvetch forages (Astragalus canadensis and Astragalus miser containing 80 and 174 soluble µmol NPA or NPOH/g forage dry matter respectively). Incubations containing 0.5 g alfalfa (Medicago sativa) were used as controls. Tubes (3 per each respective forage) were capped and incubated anaerobically (using oxygen free carbon dioxide) for 24 h at 39oC after which time amounts of total gas produced were measured via volume displacement and headspace samples were analyzed by gas chromatography to determine concentrations of hydrogen and methane. Fluid samples were analyzed by gas chromatography to measure accumulations of fermentation acids. A completely randomized analysis of variance revealed that the nitrate-containing barley and both the NPA- and the NPOH-containing milkvetches significantly decreased methane production, by > 50%, when compared to methane produced by populations incubated similarly with alfalfa (70.4 ± 3.6 µmol/ml incubation fluid). Accumulations of hydrogen, which are typically increased when methane production is inhibited, by incubations with the nitrate-containing barley and the NPA- and NPOH-containing milkvetches did not differ from accumulations observed in the alfalfa controls (0.09 ± 0.04 µmol/mL incubation fluid). Accumulations of fermentation acids produced in the incubations containing the high-nitrate barley and the NPA- and NPOH-containing milkvetches likewise did not differ from accumulations observed in incubations containing alfalfa (123.5 ± 10.8, 36.0 ± 3.0, 17.1 ± 1.5, 3.5 ± 0.3, 2.3 ± 0.2, 2.2 ± 0.2 µmol/mL incubation fluid for acetate, propionate, butyrate, valerate, isobutyrate, and isovalerate, respectively). This finding indicates the microbial populations did not compensate for the decreased methane production via compensatory changes in production of fermentative acids. Stoichiometric estimation of fermentation balance revealed that > 77% of reducing equivalents generated during fermentation of the forages were recovered in fermentation products and the recoveries did not differ between the alfalfa incubations and those with the high-nitrate barley or the NPA- or NPOH-containing milkvetches. Stoichiometric estimates of amounts of hexose fermented similarly did not differ between the nitrate-, NPA and NPOH-containing incubations and those with the alfalfa, averaging 99.6 ± 37.2 µmol hexose consumed/mL of incubation fluid. These results suggest that forages containing nitrate, NPA or NPOH may be useful to reduce methane emissions of grazing ruminants provided risks of toxicity can be effectively managed.

Keywords: nitrate, nitropropanol, nitropropionic acid, rumen methane emissions

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128 Effect of Dietary Inclusion of Moringa oleifera Leaf Meal on Blood Biochemical Changes and Lipid Profile of Vanaraja Chicken in Tropics

Authors: Kaushalendra Kumar, Abhishek Kumar, Chandra Moni, Sanjay Kumar, P. K. Singh, Ajeet Kumar

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Present study investigated the dietary inclusion of Moringa oleifera leaf meal (MOLM) on production efficiency, hemato-biochemical profile and economy of Vanaraja birds under tropical condition. Experiment was conducted for a period of 56 days on 300 Vanaraja birds randomly divided in to five different experimental groups including control of 60 birds each group replicated with 20 chicks in each replicate. T1, T2, T3, T4, and T5 were offered with 0, 5, 10, 15, and 20% Moringa oleifera leaf meal along with basal ration. All the standard managemental practices were followed during experimental period including vaccination schedule. Locally available Moringa oleifera leaves were harvested at mature stage and allowed to dry under shady and aerated conditions. Thereafter, dried leaves were milled to make a leaf meal and stored in the airtight nylon bags to avoid any possible contamination from foreign material and use for experiment. Production parameters were calculated based on the amount of feed consumed and weight gain every weeks. The body weight gain of T2 group was significantly (P < 0.05) higher side whereas T3 group was comparable with control. The feed conversion ratio for T2 group was found to be significantly (P < 0.05) lower than all other treatment groups, while none of the group was comparable with each other. At the end of the experiment blood samples were collected from birds for haematology study while serum biochemistry performed using spectrophotometer following statndard protocols. The haematological attributes were significantly (P > 0.05) not differed among the groups. However, serum biochemistry showed significant reduction (P < 0.05) of blood urea nitrogen, uric acid and creatinine level with higher level of MOLM diet, indicates better utilization of protein supplemented through MOLM. The total cholesterol and triglyceride level was declined significantly (P < 0.05) as compare to control group with increased level of MOLM in basal diet, decreasing trend of serum cholesterol noted. However, value of HDL for T3 group was highest and for T1 group was lowest but no significant difference (P < 0.05) found among the groups. It might be due to presence of β-sitosterol a bioactive compound present in MOLM which causes lowering of plasma concentration of LDL. During experiment total, LDL and VLDL level was found to be decreased significantly (P < 0.05) as compare to control group. It was observed that the production efficiency of birds significantly improved with 5% followed by 10% Moringa oleifera leaf meal among the treatment groups. However, the maximum profit per kg live weight was noted in 10 % level and least profit observed in 20% MOLM fed group. It was concluded that the dietary inclusion of MOLM improved overall performances without affecting metabolic status and effective in reducing cholesterol level reflects healthy chicken production for human consumption.

Keywords: hemato biochemistry, Moringa oleifera leaf meal, performance, Vanaraja birds

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127 Inclusion Body Refolding at High Concentration for Large-Scale Applications

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

Procedia PDF Downloads 285
126 Role of HIV-Support Groups in Mitigating Adverse Sexual Health Outcomes among HIV Positive Adolescents in Uganda

Authors: Lilian Nantume Wampande

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Group-based strategies in the delivery of HIV care have opened up new avenues not only for meaningful participation for HIV positive people but also platforms for deconstruction and reconstruction of knowledge about living with the virus. Yet the contributions of such strategies among patients who live in high risk areas are still not explored. This case study research assessed the impact of HIV support networks on sexual health outcomes of HIV positive out-of-school adolescents residing in fishing islands of Kalangala in Uganda. The study population was out-of-school adolescents living with HIV and their sexual partners (n=269), members of their households (n=80) and their health service providers (n=15). Data were collected via structured interviews, observations and focus group discussions between August 2016 and March 2017. Data was then analyzed inductively to extract key themes related to the approaches and outcomes of the groups’ activities. The study findings indicate that support groups unite HIV positive adolescents in a bid for social renegotiation to achieve change but individual constraints surpass the groups’ intentions. Some adolescents for example reported increased fear which led to failure to cope, sexual violence, self-harm and denial of status as a result of the high expectations placed on them as members of the support groups. Further investigations around this phenomenon show that HIV networks play a monotonous role as information sources for HIV positive out-of-school adolescents which limit their creativity to seek information elsewhere. Results still indicate that HIV adolescent groups recognize the complexity of long-term treatment and stay in care leading to improved immunity for the majority yet; there is still scattered evidence about how effective they are among adolescents at different phases in the disease trajectory. Nevertheless, the primary focus of developing adolescent self-efficacy and coping skills significantly address a range of disclosure difficulties and supports autonomy. Moreover, the peer techniques utilized in addition to the almost homogeneous group characteristics accelerates positive confidence, hope and belongingness. Adolescent HIV-support groups therefore have the capacity to both improve and/or worsen sexual health outcomes for a young adolescent who is out-of-school. Communication interventions that seek to increase awareness about ‘self’ should therefore be emphasized more than just fostering collective action. Such interventions should be sensitive to context and gender. In addition, facilitative support supervision done by close and trusted health care providers, most preferably Village Health Teams (who are often community elected volunteers) would help to follow-up, mentor, encourage and advise this young adolescent in matters involving sexuality and health outcomes. HIV/AIDS prevention programs have extended their efforts beyond individual focus to those that foster collective action, but programs should rekindle interpersonal level strategies to address the complexity of individual behavior.

Keywords: adolescent, HIV, support groups, Uganda

Procedia PDF Downloads 127
125 Effect of Supplementation with Fresh Citrus Pulp on Growth Performance, Slaughter Traits and Mortality in Guinea Pigs

Authors: Carlos Minguez, Christian F. Sagbay, Erika E. Ordoñez

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Guinea pigs (Cavia porcellus) play prominent roles as experimental models for medical research and as pets. However, in developing countries like South America, the Philippines, and sub-Saharan Africa, the meat of guinea pigs is an economic source of animal protein for the poor and malnourished humans because guinea pigs are mainly fed with forage and do not compete directly with human beings for food resources, such as corn or wheat. To achieve efficient production of guinea pigs, it is essential to provide insurance against vitamin C deficiency. The objective of this research was to investigate the effect of the partial replacement of alfalfa with fresh citrus pulp (Citrus sinensis) in a diet of guinea pigs on the growth performance, slaughter traits and mortality during the fattening period (between 20 and 74 days of age). A total of 300 guinea pigs were housed in collective cages of about ten animals (2 x 1 x 0.4 m) and were distributed into two completely randomized groups. Guinea pigs in both groups were fed ad libitum, with a standard commercial pellet diet (10 MJ of digestible energy/kg, 17% crude protein, 11% crude fiber, and 4.5% crude fat). Control group was supplied with fresh alfalfa as forage. In the treatment group, 30% of alfalfa was replaced by fresh citrus pulp. Growth traits, including body weight (BW), average daily gain (ADG), feed intake (FI), and feed conversion ratio (FCR), were measured weekly. On day 74, the animals were slaughtered, and slaughter traits, including live weight at slaughter (LWS), full gastrointestinal tract weight (FGTW), hot carcass weight (with head; HCW), cold carcass weight (with head; CCW), drip loss percentage (DLP) and dressing out carcass yield percentage (DCY), were evaluated. Contrasts between groups were obtained by calculated generalized least squares values. Mortality was evaluated by Fisher's exact test due to low numbers in some cells. In the first week, there were significant differences in the growth traits BW, ADG, FI, and FCR, which were superior in control group. These differences may have been due to the origin of the young guinea pigs, which, before weaning, were all raised without fresh citrus pulp, and they were not familiarized with the new supplement. In the second week, treatment group had significantly increased ADG compared with control group, which may have been the result of a process of compensatory growth. During subsequent weeks, no significant differences were observed between animals raised in the two groups. Neither were any significant differences observed across the total fattening period. No significant differences in slaughter traits or mortality rate were observed between animals from the two groups. In conclusion, although there were no significant differences in growth performance, slaughter traits, or mortality, the use of fresh citrus pulp is recommended. Fresh citrus pulp is a by-product of orange juice industry and it is cheap or free. Forage made with fresh citrus pulp could reduce about of 30 % the quantity of alfalfa in guinea pig for meat and as consequence, reduce the production costs.

Keywords: fresh citrus, growth, Guinea pig, mortality

Procedia PDF Downloads 178
124 Gene Expression Profiling of Iron-Related Genes of Pasteurella multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar

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Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.

Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes

Procedia PDF Downloads 447
123 Solutions for Food-Safe 3D Printing

Authors: Geremew Geidare Kailo, Igor Gáspár, András Koris, Ivana Pajčin, Flóra Vitális, Vanja Vlajkov

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Three-dimension (3D) printing, a very popular additive manufacturing technology, has recently undergone rapid growth and replaced the use of conventional technology from prototyping to producing end-user parts and products. The 3D Printing technology involves a digital manufacturing machine that produces three-dimensional objects according to designs created by the user via 3D modeling or computer-aided design/manufacturing (CAD/CAM) software. The most popular 3D printing system is Fused Deposition Modeling (FDM) or also called Fused Filament Fabrication (FFF). A 3D-printed object is considered food safe if it can have direct contact with the food without any toxic effects, even after cleaning, storing, and reusing the object. This work analyzes the processing timeline of the filament (material for 3D printing) from unboxing to the extrusion through the nozzle. It is an important task to analyze the growth of bacteria on the 3D printed surface and in gaps between the layers. By default, the 3D-printed object is not food safe after longer usage and direct contact with food (even though they use food-safe filaments), but there are solutions for this problem. The aim of this work was to evaluate the 3D-printed object from different perspectives of food safety. Firstly, testing antimicrobial 3D printing filaments from a food safety aspect since the 3D Printed object in the food industry may have direct contact with the food. Therefore, the main purpose of the work is to reduce the microbial load on the surface of a 3D-printed part. Coating with epoxy resin was investigated, too, to see its effect on mechanical strength, thermal resistance, surface smoothness and food safety (cleanability). Another aim of this study was to test new temperature-resistant filaments and the effect of high temperature on 3D printed materials to see if they can be cleaned with boiling or similar hi-temp treatment. This work proved that all three mentioned methods could improve the food safety of the 3D printed object, but the size of this effect variates. The best result we got was with coating with epoxy resin, and the object was cleanable like any other injection molded plastic object with a smooth surface. Very good results we got by boiling the objects, and it is good to see that nowadays, more and more special filaments have a food-safe certificate and can withstand boiling temperatures too. Using antibacterial filaments reduced bacterial colonies to 1/5, but the biggest advantage of this method is that it doesn’t require any post-processing. The object is ready out of the 3D printer. Acknowledgements: The research was supported by the Hungarian and Serbian bilateral scientific and technological cooperation project funded by the Hungarian National Office for Research, Development and Innovation (NKFI, 2019-2.1.11-TÉT-2020-00249) and the Ministry of Education, Science and Technological Development of the Republic of Serbia. The authors acknowledge the Hungarian University of Agriculture and Life Sciences’s Doctoral School of Food Science for the support in this study

Keywords: food safety, 3D printing, filaments, microbial, temperature

Procedia PDF Downloads 129
122 Revolutionary Wastewater Treatment Technology: An Affordable, Low-Maintenance Solution for Wastewater Recovery and Energy-Saving

Authors: Hady Hamidyan

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As the global population continues to grow, the demand for clean water and effective wastewater treatment becomes increasingly critical. By 2030, global water demand is projected to exceed supply by 40%, driven by population growth, increased water usage, and climate change. Currently, about 4.2 billion people lack access to safely managed sanitation services. The wastewater treatment sector faces numerous challenges, including the need for energy-efficient solutions, cost-effectiveness, ease of use, and low maintenance requirements. This abstract presents a groundbreaking wastewater treatment technology that addresses these challenges by offering an energy-saving approach, wastewater recovery capabilities, and a ready-made, affordable, and user-friendly package with minimal maintenance costs. The unique design of this ready-made package made it possible to eliminate the need for pumps, filters, airlift, and other common equipment. Consequently, it enables sustainable wastewater treatment management with exceptionally low energy and cost requirements, minimizing investment and maintenance expenses. The operation of these packages is based on continuous aeration, which involves injecting oxygen gas or air into the aeration chamber through a tubular diffuser with very small openings. This process supplies the necessary oxygen for aerobic bacteria. The recovered water, which amounts to almost 95% of the input, can be treated to meet specific quality standards, allowing safe reuse for irrigation, industrial processes, or even potable purposes. This not only reduces the strain on freshwater resources but also provides economic benefits by offsetting the costs associated with freshwater acquisition and wastewater discharge. The ready-made, affordable, and user-friendly nature of this technology makes it accessible to a wide range of users, including small communities, industries, and decentralized wastewater treatment systems. The system incorporates user-friendly interfaces, simplified operational procedures, and integrated automation, facilitating easy implementation and operation. Additionally, the use of durable materials, efficient equipment, and advanced monitoring systems significantly reduces maintenance requirements, resulting in low overall life-cycle costs and alleviating the burden on operators and maintenance personnel. In conclusion, the presented wastewater treatment technology offers a comprehensive solution to the challenges faced by the industry. Its energy-saving approach, combined with wastewater recovery capabilities, ensures sustainable resource management and enhances environmental stewardship. This affordable, ready-made, and low-maintenance package promotes broad adoption across various sectors and communities, contributing to a more sustainable future for water and wastewater management.

Keywords: wastewater treatment, energy saving, wastewater recovery, affordable package, low maintenance costs, sustainable resource management, environmental stewardship

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121 Establishing an Evidence-Based Trauma Informed Care Pathway for Survivors of Modern Slavery

Authors: I. Brezeanu, J. Mackrill, A. Cajo, C. Mogollon

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Modern Slavery is a serious crime, where often the victims are unable to leave their situation of exploitation, being controlled by threats, punishment, violence, coercion, and deception. In the UK, this term encompasses both Slavery and Human Trafficking. The number of potential victims who were referred to the National Referral Mechanism (NRM) increased exponentially in the past decade, passing from fewer than 700 potential victims referred in 2010 to more than 12.000 in 2021. Our study aims to explore how the concept of Trauma-Informed Care (TIC) approach can be adopted by services working with survivors of Modern Slavery and Trafficking (MST). Notably, in this paper, we will elaborate on how the complex needs of survivors are related to their traumatic experiences and what are the necessary steps and resources for implementing a Modern Slavery Trauma-Informed model. While there are relatively few services in the UK that have a deep understanding of the survivors’ and practitioners’ views of how trauma impacts their daily life, there is a strong need for developing services that are organised and delivered in ways that prevent retraumatisation and enable trauma survivors to engage safely with the right professionals at the right time, promoting healing through positive relationships. Such models, known as Trauma-Informed Approaches (TIAs), are seen as crucial to the empowerment of survivors, yet they remain a marginal implementation model by governments, law enforcement, judiciary, or care providers, who are frequently survivors’ first point of contact in the recovery process. In order to understand better how to provide best practice and to adopt the concept, this study is based on a multi-disciplinary approach, encompassing both theoretical perspectives and co-production. By combining qualitative and quantitative research and comparing different analysis of applied examples of TIC in the US and the UK, we gained important insights about the prevention and impact of trauma on survivors’ life. The articulation between more general expertise on Trauma-Informed Care developed by other institutions operating in the field, and the SJOG delivery, based on the Salvation Army’s Modern Slavery Victim Care and Coordination Contract (MSVCC) and the Care Quality Commission regulations, allowed to identify on one side what are the complex needs of survivors derived from their traumatic experiences, and on the other side, how could MST services prevent retraumatisation. Additional, two in-depth interviews with survivors, who receive support from one of our services at Olallo House in London, and a survey shared among all colleagues working with MST services completed the findings of the research with their personal experience and knowledge. Ultimately, we developed an evidence-based Trauma-Informed Care Pathway that aims to improve the wellbeing of survivors and to support them to live a meaningful life. The establishedpathway delivers three main outcomes belonging to the social determinants of health criteria – health and wellbeing, purpose and relationship, and covers key themes of the context of trauma, needs of individuals, and service support.

Keywords: trauma-informed care, modern slavery, human trafficking, trauma, retraumatisation

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120 Mycophenolate-Induced Disseminated TB in a PPD-Negative Patient

Authors: Megan L. Srinivas

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Individuals with underlying rheumatologic diseases such as dermatomyositis may not adequately respond to tuberculin (PPD) skin tests, creating false negative results. These illnesses are frequently treated with immunosuppressive therapy making proper identification of TB infection imperative. A 59-year-old Filipino man was diagnosed with dermatomyositis on the basis of rash, electromyography, and muscle biopsy. He was initially treated with IVIG infusions and transitioned to oral prednisone and mycophenolate. The patient’s symptoms improved on this regimen. Six months after starting mycophenolate, the patient began having fevers, night sweats, and productive cough without hemoptysis. He moved from the Philippines 5 years prior to dermatomyositis diagnosis, denied sick contacts, and was PPD negative both at immigration and immediately prior to starting mycophenolate treatment. A third PPD was negative following the onset of these new symptoms. He was treated for community-acquired pneumonia, but symptoms worsened over 10 days and he developed watery diarrhea and a growing non-tender, non-mobile mass on the left side of his neck. A chest x-ray demonstrated a cavitary lesion in right upper lobe suspicious for TB that had not been present one month earlier. Chest CT corroborated this finding also exhibiting necrotic hilar and paratracheal lymphadenopathy. Neck CT demonstrated the left-sided mass as cervical chain lymphadenopathy. Expectorated sputum and stool samples contained acid-fast bacilli (AFB), cultures showing TB bacteria. Fine-needle biopsy of the neck mass (scrofula) also exhibited AFB. An MRI brain showed nodular enhancement suspected to be a tuberculoma. Mycophenolate was discontinued and dermatomyositis treatment was switched to oral prednisone with a 3-day course of IVIG. The patient’s infection showed sensitivity to standard RIPE (rifampin, isoniazid, pyrazinamide, and ethambutol) treatment. Within a week of starting RIPE, the patient’s diarrhea subsided, scrofula diminished, and symptoms significantly improved. By the end of treatment week 3, the patient’s sputum no longer contained AFB; he was removed from isolation, and was discharged to continue RIPE at home. He was discharged on oral prednisone, which effectively addressed his dermatomyositis. This case illustrates the unreliability of PPD tests in patients with long-term inflammatory diseases such as dermatomyositis. Other immunosuppressive therapies (adalimumab, etanercept, and infliximab) have been affiliated with conversion of latent TB to disseminated TB. Mycophenolate is another immunosuppressive agent with similar mechanistic properties. Thus, it is imperative that patients with long-term inflammatory diseases and high-risk TB factors initiating immunosuppressive therapy receive a TB blood test (such as a quantiferon gold assay) prior to the initiation of therapy to ensure that latent TB is unmasked before it can evolve into a disseminated form of the disease.

Keywords: dermatomyositis, immunosuppressant medications, mycophenolate, disseminated tuberculosis

Procedia PDF Downloads 196
119 Automated Prediction of HIV-associated Cervical Cancer Patients Using Data Mining Techniques for Survival Analysis

Authors: O. J. Akinsola, Yinan Zheng, Rose Anorlu, F. T. Ogunsola, Lifang Hou, Robert Leo-Murphy

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Cervical Cancer (CC) is the 2nd most common cancer among women living in low and middle-income countries, with no associated symptoms during formative periods. With the advancement and innovative medical research, there are numerous preventive measures being utilized, but the incidence of cervical cancer cannot be truncated with the application of only screening tests. The mortality associated with this invasive cervical cancer can be nipped in the bud through the important role of early-stage detection. This study research selected an array of different top features selection techniques which was aimed at developing a model that could validly diagnose the risk factors of cervical cancer. A retrospective clinic-based cohort study was conducted on 178 HIV-associated cervical cancer patients in Lagos University teaching Hospital, Nigeria (U54 data repository) in April 2022. The outcome measure was the automated prediction of the HIV-associated cervical cancer cases, while the predictor variables include: demographic information, reproductive history, birth control, sexual history, cervical cancer screening history for invasive cervical cancer. The proposed technique was assessed with R and Python programming software to produce the model by utilizing the classification algorithms for the detection and diagnosis of cervical cancer disease. Four machine learning classification algorithms used are: the machine learning model was split into training and testing dataset into ratio 80:20. The numerical features were also standardized while hyperparameter tuning was carried out on the machine learning to train and test the data. Logistic Regression (LR), Decision Tree (DT), Random Forest (RF), and K-Nearest Neighbor (KNN). Some fitting features were selected for the detection and diagnosis of cervical cancer diseases from selected characteristics in the dataset using the contribution of various selection methods for the classification cervical cancer into healthy or diseased status. The mean age of patients was 49.7±12.1 years, mean age at pregnancy was 23.3±5.5 years, mean age at first sexual experience was 19.4±3.2 years, while the mean BMI was 27.1±5.6 kg/m2. A larger percentage of the patients are Married (62.9%), while most of them have at least two sexual partners (72.5%). Age of patients (OR=1.065, p<0.001**), marital status (OR=0.375, p=0.011**), number of pregnancy live-births (OR=1.317, p=0.007**), and use of birth control pills (OR=0.291, p=0.015**) were found to be significantly associated with HIV-associated cervical cancer. On top ten 10 features (variables) considered in the analysis, RF claims the overall model performance, which include: accuracy of (72.0%), the precision of (84.6%), a recall of (84.6%) and F1-score of (74.0%) while LR has: an accuracy of (74.0%), precision of (70.0%), recall of (70.0%) and F1-score of (70.0%). The RF model identified 10 features predictive of developing cervical cancer. The age of patients was considered as the most important risk factor, followed by the number of pregnancy livebirths, marital status, and use of birth control pills, The study shows that data mining techniques could be used to identify women living with HIV at high risk of developing cervical cancer in Nigeria and other sub-Saharan African countries.

Keywords: associated cervical cancer, data mining, random forest, logistic regression

Procedia PDF Downloads 71
118 Study of a Decentralized Electricity Market on Awaji Island

Authors: Arkadiusz P. Wójcik, Tetsuya Sato, Shin-Ichiro Shima, Mateusz Malanowski

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Over the last decades, new technologies have significantly changed the way information is transmitted and stored. Renewable energy sources have become prevalent and affordable. Cooperation of the Information and Communication Technology industry and Renewable Energy industry makes it possible to create a next generation, decentralized power grid. In this context, the study seeks to identify the wider benefits to the local Japanese economy as a result of the development of a decentralised electricity market. Our general approach aims to integrate an economic analysis (monetary appraisal of costs and benefits to society) with externalities that are not quantifiable in monetary terms (e.g. social impact, environmental impact). The study also highlights opportunities and sets out recommendations for the citizens of the island and the local government. The simulation is the scientific basis for economic impact analysis. Various types of sources of energy have been taken into account: residential wind farm, residential wind turbine, solar farm, residential solar panels and private solar farms. Analysis of local geographic and economic conditions allowed creating a customized business model. Very often farmers on Awaji Island are using crop cycle. During each cycle, one part of the field is resting and replenishing nutrients. In the next year another part of the field is resting. Portable solar panels could be freely set up in this part of the field. At the end of the crop cycle, portable solar panels would be moved to the next resting part. Because of spacious area, for a single household 500 square meters of portable solar panels has been proposed and simulated. The devised simulation shows that the Rate of Return on Investment for solar panels, which are on the island, could reach up to 37.21%. Supposing that about 20% of households install solar panels they could produce 49.11% of the electric energy consumed by households on the island. The analysis shows that rest of the energy supply can be produced by currently existing one huge solar farm and two wind farms to meet 97.59% of demand on electricity for households on the island. Although there are more than 7,000 agricultural fields on the island, young people tend to avoid agricultural work and prefer to move from the island to big cities, live there in little mansions and work until late night. The business model proposed in this study could increase farmer’s monthly income by ¥200,000 - ¥300,000 (1,600 euro – 2,400 euro). Young people could work less and have a higher standard of living than in a city. Creation of a decentralized electricity market can unlock significant benefits in other industries (e.g. electric vehicles), providing a welcome boost to economic growth, jobs and quality of life.

Keywords: digital twin, Matlab, model-based systems engineering, simulink, smart grid, systems engineering

Procedia PDF Downloads 96
117 Urban Security through Urban Transformation: Case of Saraycik District

Authors: Emir Sunguroglu, Merve Sunguroglu, Yesim Aliefendioglu, Harun Tanrivermis

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Basic human needs range from physiological needs such as food, water and shelter to safety needs such as security, protection from natural disasters and even urban terrorism which are extant and not fulfilled even in urban areas where people live civilly in large communities. These basic needs when arose in urban life lead to a different kind of crime set defined as urban crimes. Urban crimes mostly result from differences between socioeconomic conditions in society. Income inequality increases tendency towards urban crimes. Especially in slum areas and suburbs, urban crimes not only threaten public security but they also affect deliverance of public services. It is highlighted that, construction of urban security against problems caused by urban crimes is not only achieved by involvement of urban security in security of the community but also comprises juridical development and staying above a level of legal standards concurrently. The idea of urban transformation emerged as interventions to demolishment and rebuilding of built environment to solve the unhealthy urban environment, inadequate infrastructure and socioeconomic problems came up during the industrialization process. Considering the probability of urbanization process driving citizens to commit crimes, The United Nations Commission on Human Security’s focus on this theme is conferred to be a proper approach. In this study, the analysis and change in security before, through and after urban transformation, which is one of the tools related to urbanization process, is strived to be discussed through the case of Sincan County Saraycik District. The study also aims to suggest improvements to current legislation on public safety, urban resilience, and urban transformation. In spite of Saraycik District residing in a developing County in Ankara, Turkey, from urbanization perspective as well as socioeconomic and demographic indicators the District exhibits a negative view throughout the County and the country. When related to the county, rates of intentional harm reports, burglary reports, the offense of libel and threat reports and narcotic crime reports are higher. The District is defined as ‘crime hotspot’. Interviews with residents of Saraycik claim that the greatest issue of the neighborhood is Public Order and Security (82.44 %). The District becomes prominent with negative aspects, especially with the presence of unlicensed constructions, occurrence of important social issues such as crime and insecurity and complicated lives of inhabitants from poverty and low standard conditions of living. Additionally, the social structure and demographic properties and crime and insecurity of the field have been addressed in this study. Consequently, it is claimed that urban crime rates were related to level of education, employment and household income, poverty trap, physical condition of housing and structuration, accessibility of public services, security, migration, safety in terms of disasters and emphasized that urban transformation is one of the most important tools in order to provide urban security.

Keywords: urban security, urban crimes, urban transformation, Saraycik district

Procedia PDF Downloads 288
116 A User-Side Analysis of the Public-Private Partnership: The Case of the New Bundang Subway Line in South Korea

Authors: Saiful Islam, Deuk Jong Bae

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The purpose of this study is to examine citizen satisfaction and competitiveness of a Public Private Partnership project. The study focuses on PPP in the transport sector and investigates the New Bundang Subway Line (NBL) in South Korea as the object of a case study. Most PPP studies are dominated by the study of public and private sector interests, which are classified in to three major areas comprising of policy, finance, and management. This study will explore the user perspective by assessing customer satisfaction upon NBL cost and service quality, also the competitiveness of NBL compared to other alternative transport modes which serve the Jeongja – Gangnam trip or vice versa. The regular Bundang Subway Line, New Bundang Subway Line, bus and private vehicle are selected as the alternative transport modes. The study analysed customer satisfaction of NBL and citizen’s preference of alternative transport modes based on a survey in Bundang district, South Korea. Respondents were residents and employees who live or work in Bundang city, and were divided into the following areas Pangyo, Jeongjae – Sunae, Migeun – Ori – Jukjeon, and Imae – Yatap – Songnam. The survey was conducted in January 2015 for two weeks, and 753 responses were gathered. By applying the Hedonic Utility approach, the factors which affect the frequency of using NBL were found to be overall customer satisfaction, convenience of access, and the socio economic demographic of the individual. In addition, by applying the Analytic Hierarchy Process (AHP) method, criteria factors influencing the decision to select alternative transport modes were identified. Those factors, along with the author judgement of alternative transport modes, and their associated criteria and sub-criteria produced a priority list of user preferences regarding their alternative transport mode options. The study found that overall the regular Bundang Subway Line (BL), which was built and operated under a conventional procurement method was selected as the most preferable transport mode due to its cost competitiveness. However, on the sub-criteria level analysis, the NBL has competitiveness on service quality, particularly on journey time. By conducting a sensitivity analysis, the NBL can become the first choice of transport by increasing the NBL’s degree of weight associated with cost by 0,05. This means the NBL would need to reduce either it’s fare cost or transfer fee, or combine those two cost components to reduce the total of the current cost by 25%. In addition, the competitiveness of NBL also could be obtained by increasing NBL convenience through escalating access convenience such as constructing an additional station or providing more access modes. Although these convenience improvements would require a few extra minutes of journey time, the user found this to be acceptable. The findings and policy suggestions can contribute to the next phase of NBL development, showing that consideration should be given to the citizen’s voice. The case study results also contribute to the literature of PPP projects specifically from a user side perspective.

Keywords: public private partnership, customer satisfaction, public transport, new Bundang subway line

Procedia PDF Downloads 340
115 Prevalence, Antimicrobial Susceptibility Pattern and Public Health Significance for Staphylococcus aureus of Isolated From Raw Red Meat at Butchery and Abattoir House in Mekelle, Northern Ethiopia

Authors: Haftay Abraha Tadesse

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Background: Staphylococcus is a genus of worldwide distributed bacteria correlated to several infectious of different sites in human and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. Objective: The objective of this study was to determine the isolates, antimicrobial susceptibility patterns and public health significance for Staphylococcus aureus in raw meat from butchery and abattoir houses of Mekelle, Northern Ethiopia. Methodology: A cross-sectional study was conducted from April to October 2019. Sociodemographic data and public health significance were collected using predesigned questionnaire. The raw meat samples were collected aseptically in the butchery and abattoir houses and transported using ice box to Mekelle University, College of Veterinary Sciences for isolating and identification of Staphylococcus aureus. Antimicrobial susceptibility tests were determined by disc diffusion method. Data obtained were cleaned and entered in to STATA 22.0 and logistic regression model with odds ratio were calculated to assess the association of risk factors with bacterial contamination. P-value < 0.05 was considered as statistically significant. Results: In present study, 88 out of 250 (35.2%) were found to be contamination with Staphylococcus aureus. Among the raw meat specimens to be positivity rate of Staphylococcus aureus were 37.6% (n=47) and (32.8% (n=41), butchery and abattoir houses, respectively. Among the associated risk factories not using gloves reduces risk was found to (AOR=0.222; 95% CI: 0.104-0.473), Strict Separation b/n clean & dirty (AOR= 1.37; 95% CI: 0.66-2.86) and poor habit of hand washing (AOR=1.08; 95%CI: 0.35-3.35) were found to be statistically significant and ha ve associated with Staphylococcus aureus contamination. All isolates thirty sevevn of Staphyloco ccus aureus were checked displayed (100%) sensitive to doxycycline, trimethoprim, gentamicin, sulphamethoxazole, amikacin, CN, Co trimoxazole and nitrofurantoi. whereas the showed resistance of cefotaxime (100%), ampicillin (87.5%), Penicillin (75%), B (75%), and nalidixic acid (50%) from butchery houses. On the other hand, all isolates of Staphylococcus aur eu isolate 100% (n= 10) showed sensitive chloramphenicol, gentamicin and nitrofurantoin whereas the showed 100% resistance of Penicillin, B, AMX, ceftriaxone, ampicillin and cefotaxime from abattoirs houses. The overall multi drug resistance pattern for Staphylococcus aureus were 90% and 100% of butchery and abattoirs houses, respectively. Conclusion: 35.3% Staphylococcus aureus isolated were recovered from the raw meat samples collected from the butchery and abattoirs houses. More has to be done in the developed of hand washing behavior, and availability of safe water in the butchery houses to reduce burden of bacterial contamination. The results of the present finding highlight the need to implement protective measures against the levels of food contamination and alternative drug options. The development of antimicrobial resistance is nearly always as a result of repeated therapeutic and/or indiscriminate use of them. Regular antimicrobial sensitivity testing helps to select effective antibiotics and to reduce the problems of drug resistance development towards commonly used antibiotics. Key words: abattoir houses, antimicrobial resistance, butchery houses, Ethiopia,

Keywords: abattoir houses, antimicrobial resistance, butchery houses, Ethiopia, staphylococcus aureuse, MDR

Procedia PDF Downloads 53
114 Potential of Dredged Material for CSEB in Building Structure

Authors: BoSheng Liu

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The research goal is to re-image a locally-sourced waste product as abuilding material. The author aims to contribute to the compressed stabilized earth block (CSEB) by investigating the promising role of dredged material as an alternative building ingredient in the production of bricks and tiles. Dredged material comes from the sediment deposited near the shore or downstream, where the water current velocity decreases. This sediment needs to be dredged to provide water transportation; thus, there are mounds of the dredged material stored at bay. It is the interest of this research to reduce the filtered un-organic soil in the production of CSEB and replace it with locally dredged material from the Atchafalaya River in Morgan City, Louisiana. Technology and mechanical innovations have evolved the traditional adobe production method, which mixes the soil and natural fiber into molded bricks, into chemically stabilized CSEB made by compressing the clay mixture and stabilizer in a compression chamber with particular loads. In the case of dredged material CSEB (DM-CSEB), cement plays an essential role as the bending agent contributing to the unit strength while sustaining the filtered un-organic soil. Each DM-CSEB unit is made in a compression chamber with 580 PSI (i.e., 4 MPa) force. The research studied the cement content from 5% to 10% along with the range of dredged material mixtures, which differed from 20% to 80%. The material mixture content affected the DM-CSEB's strength and workability during and after its compression. Results indicated two optimal workabilities of the mixture: 27% fine clay content and 63% dredged material with 10% cement, or 28% fine clay content, and 67% dredged material with 5% cement. The final product of DM-CSEB emitted between 10 to 13 times fewer carbon emissions compared to the conventional fired masonry structure. DM-CSEB satisfied the strength requirement given by the ASTM C62 and ASTM C34 standards for construction material. One of the final evaluations tested and validated the material performance by designing and constructing an architectural, conical tile-vault prototype that was 28" by 40" by 24." The vault utilized a computational form-finding approach to generate the form's geometry, which optimized the correlation between the vault geometry and structural load distribution. A series of scaffolding was deployed to create the framework for the tile-vault construction. The final tile-vault structure was made from 2 layers of DM-CSEB tiles jointed by mortar, and the construction of the structure used over 110 tiles. The tile-vault prototype was capable of carrying over 400 lbs of live loads, which further demonstrated the dredged material feasibility as a construction material. The presented case study of Dredged Material Compressed Stabilized Earth Block (DM-CSEB) provides the first impression of dredged material in the clayey mixture process, structural performance, and construction practice. Overall, the approach of integrating dredged material in building material can be feasible, regionally sourced, cost-effective, and environment-friendly.

Keywords: dredged material, compressed stabilized earth block, tile-vault, regionally sourced, environment-friendly

Procedia PDF Downloads 103
113 Glycyrrhizic Acid Inhibits Lipopolysaccharide-Stimulated Bovine Fibroblast-Like Synoviocyte, Invasion through Suppression of TLR4/NF-κB-Mediated Matrix Metalloproteinase-9 Expression

Authors: Hosein Maghsoudi

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Rheumatois arthritis (RA) is progressive inflammatory autoimmune diseases that primarily affect the joints, characterized by synovial hyperplasia and inflammatory cell infiltration, deformed and painful joints, which can lead tissue destruction, functional disability systemic complications, and early dead and socioeconomic costs. The cause of rheumatoid arthritis is unknown, but genetic and environmental factors are contributory and the prognosis is guarded. However, advances in understanding the pathogenesis of the disease have fostered the development of new therapeutics, with improved outcomes. The current treatment strategy, which reflects this progress, is to initiate aggressive therapy soon after diagnosis and to escalate the therapy, guided by an assessment of disease activity, in pursuit of clinical remission. The pathobiology of RA is multifaceted and involves T cells, B cells, fibroblast-like synoviocyte (FLSc) and the complex interaction of many pro-inflammatory cytokine. Novel biologic agents that target tumor necrosis or interlukin (IL)-1 and Il-6, in addition T- and B-cells inhibitors, have resulted in favorable clinical outcomes in patients with RA. Despite this, at least 30% of RA patients are résistance to available therapies, suggesting novel mediators should be identified that can target other disease-specific pathway or cell lineage. Among the inflammatory cell population that might participated in RA pathogenesis, FLSc are crucial in initiaing and driving RA in concert of cartilage and bone by secreting metalloproteinase (MMPs) into the synovial fluid and by direct invasion into extracellular matrix (ECM), further exacerbating joint damage. Invasion of fibroblast-like synoviocytes (FLSc) is critical in the pathogenesis of rheumatoid-arthritis. The metalloproteinase (MMPs) and activator of Toll-like receptor 4 (TLR4)/nuclear factor- κB pthway play a critical role in RA-FLS invasion induced by lipopolysaccharide (LPS). The present study aimed to explore the anti-invasion activity of Glycyrrhizic Acid as a pharmacologically safe phytochemical agent with potent anti-inflammatory properties on IL-1beta and TNF-alpha signalling pathways in Bovine fibroblast-like synoviocyte ex- vitro, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Results showed that Glycyrrhizic Acid suppressed LPS-stimulated bovine FLS migration and invasion by inhibition MMP-9 expression and activity. In addition our results revealed that Glycyrrhizic Acid inhibited the transcriptional activity of MMP-9 by suppression the nbinding activity of NF- κB in the MMP-9 promoter pathway. The extract of licorice (Glycyrrhiza glabra L.) has been widely used for many centuries in the traditional Chinese medicine as native anti-allergic agent. Glycyrrhizin (GL), a triterpenoidsaponin, extracted from the roots of licorice is the most effective compound for inflammation and allergic diseases in human body. The biological and pharmacological studies revealed that GL possesses many pharmacological effects, such as anti-inflammatory, anti-viral and liver protective effects, and the biological effects, such as induction of cytokines (interferon-γ and IL-12), chemokines as well as extrathymic T and anti-type 2 T cells. GL is known in the traditional Chinese medicine for its anti-inflammatory effect, which is originally described by Finney in 1959. The mechanism of the GL-induced anti-inflammatory effect is based on different pathways of the GL-induced selective inhibition of the prostaglandin E2 production, the CK-II- mediated activation of both GL-binding lipoxygenas (gbLOX; 17) and PLA2, an anti-thrombin action of GL and production of the reactive oxygen species (ROS; GL exerts liver protection properties by inhibiting PLA2 or by the hydroxyl radical trapping action, leading to the lowering of serum alanine and aspartate transaminase levels. The present study was undertaken to examine the possible mechanism of anti-inflammatory properties GL on IL-1beta and TNF-alpha signalling pathways in bovine fibroblast-like synoviocyte ex-vivo, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Our results clearly showed that treatment of bovine fibroblast-like synoviocyte with GL suppressed LPS-induced cell migration and invasion. Furthermore, it revealed that GL inhibited the transcription activity of MMP-9 by suppressing the binding activity of NF-κB in the MM-9 promoter. MMP-9 is an important ECM-degrading enzyme and overexpression of MMPs in important of RA-FLSs. LPS can stimulate bovine FLS to secret MMPs, and this induction is regulated at the transcription and translational levels. In this study, LPS treatment of bovine FLS caused an increase in MMP-2 and MMP-9 levels. The increase in MMP-9 expression and secretion was inhibited by ex- vitro. Furthermore, these effects were mimicked by MMP-9 siRNA. These result therefore indicate the the inhibition of LPS-induced bovine FLS invasion by GL occurs primarily by inhibiting MMP-9 expression and activity. Next we analyzed the functional significance of NF-κB transcription of MMP-9 activation in Bovine FLSs. Results from EMSA showed that GL suppressed LPS-induced NF-κB binding to the MMP-9 promotor, as NF-κB regulates transcriptional activation of multiple inflammatory cytokines, we predicted that GL might target NF-κB to suppress MMP-9 transcription by LPS. Myeloid differentiation-factor 88 (MyD88) and TIR-domain containing adaptor protein (TIRAP) are critical proteins in the LPS-induced NF-κB and apoptotic signaling pathways, GL inhibited the expression of TLR4 and MYD88. These results demonstrated that GL suppress LPS-induced MMP-9 expression through the inhibition of the induced TLR4/NFκB signaling pathway. Taken together, our results provide evidence that GL exerts anti-inflammatory effects by inhibition LPS-induced bovine FLSs migration and invasion, and the mechanisms may involve the suppression of TLR4/NFκB –mediated MMP-9 expression. Although further work is needed to clarify the complicated mechanism of GL-induced anti-invasion of bovine FLSs, GL might be used as a further anti-invasion drug with therapeutic efficacy in the treatment of immune-mediated inflammatory disease such as RA.

Keywords: glycyrrhizic acid, bovine fibroblast-like synoviocyte, tlr4/nf-κb, metalloproteinase-9

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112 Isoflavonoid Dynamic Variation in Red Clover Genotypes

Authors: Andrés Quiroz, Emilio Hormazábal, Ana Mutis, Fernando Ortega, Loreto Méndez, Leonardo Parra

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Red clover root borer, Hylastinus obscurus Marsham (Coleoptera: Curculionidae), is the main insect pest associated to red clover, Trifolium pratense L. An average of 1.5 H. obscurus per plant can cause 5.5% reduction in forage yield in pastures of two to three years old. Moreover, insect attack can reach 70% to 100% of the plants. To our knowledge, there is no a chemical strategy for controlling this pest. Therefore alternative strategies for controlling H. obscurus are a high priority for red clover producers. One of this alternative is related to the study of secondary metabolites involved in intrinsic chemical defenses developed by plants, such as isoflavonoids. The isoflavonoids formononetin and daidzein have elicited an antifeedant and phagostimult effect on H. obscurus respectively. However, we do not know how is the dynamic variation of these isoflavonoids under field conditions. The main objective of this work was to evaluate the variation of the antifeedant isoflavonoids formononetin, the phagostimulant isoflavonoids daidzein, and their respective glycosides over time in different ecotypes of red clover. Fourteen red clover ecotypes (8 cultivars and 6 experimental lines), were collected at INIA-Carillanca (La Araucanía, Chile). These plants were established in October 2015 under irrigated conditions. The cultivars were distributed in a randomized complete block with three replicates. The whole plants were sampled in four times: 15th October 2016, 12th December 2016, 27th January 2017 and 16th March 2017 with sufficient amount of soil to avoid root damage. A polar fraction of isoflavonoid was obtained from 20 mg of lyophilized root tissue extracted with 2 mL of 80% MeOH for 16 h using an orbital shaker in the dark at room temperature. After, an aliquot of 1.4 mL of the supernatant was evaporated, and the residue was resuspended in 300 µL of 45% MeOH. The identification and quantification of isoflavonoid root extracts were performed by the injection of 20 µL into a Shimadzu HPLC equipped with a C-18 column. The sample was eluted with a mobile phase composed of AcOH: H₂O (1:9 v/v) as solvent A and CH₃CN as solvent B. The detection was performed at 260 nm. The results showed that the amount of aglycones was higher than the respective glycosides. This result is according to the biosynthetic pathway of flavonoids, where the formation of glycoside is further to the glycosides biosynthesis. The amount of formononetin was higher than daidzein. In roots, where H. obscurus spent the most part of its live cycle, the highest content of formononetin was found in G 27, Pawera, Sabtoron High, Redqueli-INIA and Superqueli-INIA cvs. (2.1, 1.8, 1.8, 1.6 and 1.0 mg g⁻¹ respectively); and the lowest amount of daidzein were found Superqueli-INIA (0.32 mg g⁻¹) and in the experimental line Sel Syn Int4 (0.24 mg g⁻¹). This ecotype showed a high content of formononetin (0.9 mg g⁻¹). This information, associated with cultural practices, could help farmers and breeders to reduce H. obscurus in grassland, selecting ecotypes with high content of formononetin and low amount of daidzein in the roots of red clover plants. Acknowledgements: FONDECYT 1141245 and 11130715.

Keywords: daidzein, formononetin, isoflavonoid glycosides, trifolium pratense

Procedia PDF Downloads 205
111 Isolation and Characterization of a Narrow-Host Range Aeromonas hydrophila Lytic Bacteriophage

Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

Procedia PDF Downloads 156
110 Floating Building Potential for Adaptation to Rising Sea Levels: Development of a Performance Based Building Design Framework

Authors: Livia Calcagni

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Most of the largest cities in the world are located in areas that are vulnerable to coastal erosion and flooding, both linked to climate change and rising sea levels (RSL). Nevertheless, more and more people are moving to these vulnerable areas as cities keep growing. Architects, engineers and policy makers are called to rethink the way we live and to provide timely and adequate responses not only by investigating measures to improve the urban fabric, but also by developing strategies capable of planning change, exploring unusual and resilient frontiers of living, such as floating architecture. Since the beginning of the 21st century we have seen a dynamic growth of water-based architecture. At the same time, the shortage of land available for urban development also led to reclaim the seabed or to build floating structures. In light of these considerations, time is ripe to consider floating architecture not only as a full-fledged building typology but especially as a full-fledged adaptation solution for RSL. Currently, there is no global international legal framework for urban development on water and there is no structured performance based building design (PBBD) approach for floating architecture in most countries, let alone national regulatory systems. Thus, the research intends to identify the technological, morphological, functional, economic, managerial requirements that must be considered in a the development of the PBBD framework conceived as a meta-design tool. As it is expected that floating urban development is mostly likely to take place as extension of coastal areas, the needs and design criteria are definitely more similar to those of the urban environment than of the offshore industry. Therefor, the identification and categorization of parameters takes the urban-architectural guidelines and regulations as the starting point, taking the missing aspects, such as hydrodynamics, from the offshore and shipping regulatory frameworks. This study is carried out through an evidence-based assessment of performance guidelines and regulatory systems that are effective in different countries around the world addressing on-land and on-water architecture as well as offshore and shipping industries. It involves evidence-based research and logical argumentation methods. Overall, this paper highlights how inhabiting water is not only a viable response to the problem of RSL, thus a resilient frontier for urban development, but also a response to energy insecurity, clean water and food shortages, environmental concerns and urbanization, in line with Blue Economy principles and the Agenda 2030. Moreover, the discipline of architecture is presented as a fertile field for investigating solutions to cope with climate change and its effects on life safety and quality. Future research involves the development of a decision support system as an information tool to guide the user through the decision-making process, emphasizing the logical interaction between the different potential choices, based on the PBBD.

Keywords: adaptation measures, floating architecture, performance based building design, resilient architecture, rising sea levels

Procedia PDF Downloads 78
109 Wildfire Risk and Biodiversity Management: Understanding Perceptions and Preparedness

Authors: Emily Moskwa, Delene Weber, Jacob Arnold, Guy M. Robinson, Douglas K. Bardsley

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Management strategies to reduce the risks to human life and property from wildfire are key contemporary concerns, with a growing literature exploring these issues from a social research perspective. Efforts range from narrowly focused examinations, such as comparing the level of community support for vegetation clearance with that of controlled burning, to broader considerations of what constitutes effective fire management policy and education campaigns. However, little analysis is available that integrates the social component of risk mitigation and the influence of educational materials with the biodiversity conservation strategies so often needed in fire-prone ecosystems found on the periphery of urban areas. Indeed many communities living on the fringe of Australian cities face major issues relating to an increased risk of wildfire events and a decline in local biodiversity. Inadequate policy and planning, and a lack of awareness or information, exacerbate this risk. This has brought forward an emerging governance challenge that requires the mitigation of wildfire risk while simultaneously supporting improved conservation practices in these urban-fringe areas. Focusing on the perceptions and experiences of residents of the Lower Eyre Peninsula in South Australia, this study analyses data collected from a series of semi-structured interviews with landholders (n=20) living in rural and urban-fringe areas surrounding the city of Port Lincoln, a city with a growing population and one that has faced a number of very large fires in recent years. In South Australia, new policies have assigned increased responsibility on individual landholders to manage their land and prepare themselves for a wildfire event, potentially to the detriment of the surrounding native vegetation. Our findings indicate the value of gaining a more nuanced understanding of the perceptions and behaviours of landholders living in areas of high fire risk, who often choose to live there in order to be close to the natural environment. Many interviewees demonstrated a high awareness of wildfire risk as a result of their past experience with fire, and the majority considered themselves to be well-prepared in the event of a future fire. Community interactions and educational programs were found to be effective in raising awareness of risk; however, negative trust relationships with government authorities and low exposure to information concerning biodiversity resulted in an overall misunderstanding of the relationship between risk mitigation and biodiversity protection. The study offers insights into how catastrophic fires are reframing perceptions of what constitutes effective vegetation management. It provides recommendations to assist with the development of education strategies that concurrently address wildfire management and biodiversity conservation, and contribute towards environmentally-informed and risk conscious governance.

Keywords: biodiversity conservation, risk, peri-urban planning, wildfire management

Procedia PDF Downloads 236
108 Bacterial Community Diversity in Soil under Two Tillage Systems

Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis

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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis

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107 Addressing Housing Issue at Regional Level Planning: A Case Study of Mumbai Metropolitan Region

Authors: Bhakti Chitale

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Mumbai city, which is the business capital of India and one of the most crowded cities in the world, holds the biggest slum in Asia. The Mumbai Metropolitan Region (MMR) occupies an area of 4035 sq.km. with a population of 22.8 million people. This population is mostly urban with 91% of this population living in areas of Municipal Corporations and Councils. Another 3% live in Census Towns. The region has 9 Municipal Corporations, 8 Municipal councils, and around 1000 villages. On the one hand MMR reflects the highest contribution to the Nations overall economy and on the other hand it shows the horrible and intolerable picture of about 2 million people, who are living in slums/without even slum with totally unhygienic conditions and with total loss of hope. The generations are about to get affected adversely if the solution is not worked out. This study is an attempt towards working out the solution. Mumbai Metropolitan Region Development Authority (MMRDA) is state government's authority, specially formed to govern the development of MMR. MMRDA is engaged in long term planning, promotion of new growth centres, implementation of strategic projects and financing infrastructure development. While preparing the master plan for MMR for next 20 years MMRDA conducted a detail study regarding Housing scenario in MMR and possible options for improvement. The author was the in charge officer for the said assignment. This paper puts light on the interesting outcomes of the research study, which ranges from the adverse effects of government policies, automatic responses of housing market, effects on planning processes, and overall changing needs of housing patterns in the world due to changes in the social mechanism. It alarms the urban planners who usually focus on smart infrastructure development, about allied future dangers. This housing study will explain the complexities, realities and needs of innovations in the housing policies all over the world. The paper will explain further few success stories and failure stories of government initiatives with reasons. It gives the clear idea about the differences in needs of housing for people from different economic groups and direct and indirect market pressures on low cost housing. Magical phenomenon came in front like a large percentage of vacant houses is present in spite of the huge need. Housing market gets affected by the developments or any other physical and financial changes taking place in the nearby areas or cities, also by changes in cities which are located far from the region and also by the international investments or policy changes. Instead of just depending on governments actions in case of generation of affordable housing, it becomes equally important to make the housing markets automatically generate such stock and still make them sustainable is the aim of all the movement. In summary, we may say that the paper will sequentially elaborate the complete dynamics of housing in one of the most crowded urban area in the world that is Mumbai Metropolitan Region, with a lot of data, analysis, case studies, and recommendations.

Keywords: Mumbai India, slum housing, region planning, market recommendations

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106 The Impact of Developing an Educational Unit in the Light of Twenty-First Century Skills in Developing Language Skills for Non-Arabic Speakers: A Proposed Program for Application to Students of Educational Series in Regular Schools

Authors: Erfan Abdeldaim Mohamed Ahmed Abdalla

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The era of the knowledge explosion in which we live requires us to develop educational curricula quantitatively and qualitatively to adapt to the twenty-first-century skills of critical thinking, problem-solving, communication, cooperation, creativity, and innovation. The process of developing the curriculum is as significant as building it; in fact, the development of curricula may be more difficult than building them. And curriculum development includes analyzing needs, setting goals, designing the content and educational materials, creating language programs, developing teachers, applying for programmes in schools, monitoring and feedback, and then evaluating the language programme resulting from these processes. When we look back at the history of language teaching during the twentieth century, we find that developing the delivery method is the most crucial aspect of change in language teaching doctrines. The concept of delivery method in teaching is a systematic set of teaching practices based on a specific theory of language acquisition. This is a key consideration, as the process of development must include all the curriculum elements in its comprehensive sense: linguistically and non-linguistically. The various Arabic curricula provide the student with a set of units, each unit consisting of a set of linguistic elements. These elements are often not logically arranged, and more importantly, they neglect essential points and highlight other less important ones. Moreover, the educational curricula entail a great deal of monotony in the presentation of content, which makes it hard for the teacher to select adequate content; so that the teacher often navigates among diverse references to prepare a lesson and hardly finds the suitable one. Similarly, the student often gets bored when learning the Arabic language and fails to fulfill considerable progress in it. Therefore, the problem is not related to the lack of curricula, but the problem is the development of the curriculum with all its linguistic and non-linguistic elements in accordance with contemporary challenges and standards for teaching foreign languages. The Arabic library suffers from a lack of references for curriculum development. In this paper, the researcher investigates the elements of development, such as the teacher, content, methods, objectives, evaluation, and activities. Hence, a set of general guidelines in the field of educational development were reached. The paper highlights the need to identify weaknesses in educational curricula, decide the twenty-first-century skills that must be employed in Arabic education curricula, and the employment of foreign language teaching standards in current Arabic Curricula. The researcher assumes that the series of teaching Arabic to speakers of other languages in regular schools do not address the skills of the twenty-first century, which is what the researcher tries to apply in the proposed unit. The experimental method is the method of this study. It is based on two groups: experimental and control. The development of an educational unit will help build suitable educational series for students of the Arabic language in regular schools, in which twenty-first-century skills and standards for teaching foreign languages will be addressed and be more useful and attractive to students.

Keywords: curriculum, development, Arabic language, non-native, skills

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105 Purple Spots on Historical Parchments: Confirming the Microbial Succession at the Basis of Biodeterioration

Authors: N. Perini, M. C. Thaller, F. Mercuri, S. Orlanducci, A. Rubechini, L. Migliore

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The preservation of cultural heritage is one of the major challenges of today’s society, because of the fundamental right of future generations to inherit it as the continuity with their historical and cultural identity. Parchments, consisting of a semi-solid matrix of collagen produced from animal skin (i.e., sheep or goats), are a significant part of the cultural heritage, being used as writing material for many centuries. Due to their animal origin, parchments easily undergo biodeterioration. The most common biological damage is characterized by isolated or coalescent purple spots that often leads to the detachment of the superficial layer and the loss of the written historical content of the document. Although many parchments with the same biodegradative features were analyzed, no common causative agent has been found so far. Very recently, a study was performed on a purple-damaged parchment roll dated back 1244 A.D, the A.A. Arm. I-XVIII 3328, belonging to the oldest collection of the Vatican Secret Archive (Fondo 'Archivum Arcis'), by comparing uncolored undamaged and purple damaged areas of the same document. As a whole, the study gave interesting results to hypothesize a model of biodeterioration, consisting of a microbial succession acting in two main phases: the first one, common to all the damaged parchments, is characterized by halophilic and halotolerant bacteria fostered by the salty environment within the parchment maybe induced by bringing of the hides; the second one, changing with the individual history of each parchment, determines the identity of its colonizers. The design of this model was pivotal to this study, performed by different labs of the Tor Vergata University (Rome, Italy), in collaboration with the Vatican Secret Archive. Three documents, belonging to a collection of dramatically damaged parchments archived as 'Faldone Patrizi A 19' (dated back XVII century A.D.), were analyzed through a multidisciplinary approach, including three updated technologies: (i) Next Generation Sequencing (NGS, Illumina) to describe the microbial communities colonizing the damaged and undamaged areas, (ii) RAMAN spectroscopy to analyze the purple pigments, (iii) Light Transmitted Analysis (LTA) to evaluate the kind and entity of the damage to native collagen. The metagenomic analysis obtained from NGS revealed DNA sequences belonging to Halobacterium salinarum mainly in the undamaged areas. RAMAN spectroscopy detected pigments within the purple spots, mainly bacteriorhodopsine/rhodopsin-like pigments, a purple transmembrane protein containing retinal and present in Halobacteria. The LTA technique revealed extremely damaged collagen structures in both damaged and undamaged areas of the parchments. In the light of these data, the study represents a first confirmation of the microbial succession model described above. The demonstration of this model is pivotal to start any possible new restoration strategy to bring back historical parchments to their original beauty, but also to open opportunities for intervention on a huge amount of documents.

Keywords: biodeterioration, parchments, purple spots, ecological succession

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104 Hear Me: The Learning Experience on “Zoom” of Students With Deafness or Hard of Hearing Impairments

Authors: H. Weigelt-Marom

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Over the years and up to the arousal of the COVID-19 pandemic, deaf or hard of hearing students studying in higher education institutions, participated lectures on campus using hearing aids and strategies adapted for frontal learning in a classroom. Usually, these aids were well known to them from their earlier study experience in school. However, the transition to online lessons, due to the latest pandemic, led deaf or hard of hearing students to study outside of their physical, well known learning environment. The change of learning environment and structure rose new challenges for these students. The present study examined the learning experience, limitations, challenges and benefits regarding learning online with lecture and classmates via the “Zoom” video conference program, among deaf or hard of hearing students in academia setting. In addition, emotional and social aspects related to learning in general versus the “Zoom” were examined. The study included 18 students diagnosed as deaf or hard of hearing, studying in various higher education institutions in Israel. All students had experienced lessons on the “Zoom”. Following allocation of the group study by the deaf and hard of hearing non-profit organization “Ma’agalei Shema”, and receiving the participants inform of consent, students were requested to answer a google form questioner and participate in an interview. The questioner included background information (e.g., age, year of studying, faculty etc.), level of computer literacy, and level of hearing and forms of communication (e.g., lip reading, sign language etc.). The interviews included a one on one, semi-structured, in-depth interview, conducted by the main researcher of the study (interview duration: up to 60 minutes). The interviews were held on “ZOOM” using specific adaptations for each interviewee: clear face screen of the interviewer for lip and face reading, and/ or professional sign language or live text transcript of the conversation. Additionally, interviewees used their audio devices if needed. Questions regarded: learning experience, difficulties and advantages studying using “Zoom”, learning in a classroom versus on “Zoom”, and questions concerning emotional and social aspects related to learning. Thematic analysis of the interviews revealed severe difficulties regarding the ability of deaf or hard of hearing students to comprehend during ”Zoom“ lessons without adoptive aids. For example, interviewees indicated difficulties understanding “Zoom” lessons due to their inability to use hearing devices commonly used by them in the classroom (e.g., FM systems). 80% indicated that they could not comprehend “Zoom” lessons since they could not see the lectures face, either because lectures did not agree to open their cameras or, either because they did not keep a straight forward clear face appearance while teaching. However, not all descriptions regarded learning via the “zoom” were negative. For example, 20% reported the recording of “Zoom” lessons as a main advantage. Enabling then to repeatedly watch the lessons at their own pace, mostly assisted by friends and family to translate the audio output into an accessible input. These finding and others regarding the learning experience of the group study on the “Zoom”, as well as their recommendation to enable deaf or hard of hearing students to study inclusively online, will be presented at the conference.

Keywords: deaf or hard of hearing, learning experience, Zoom, qualitative research

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103 Performance of a Lytic Bacteriophage Cocktail against Pseudomonas aeruginosa in Conditions That Simulate the Cystic Fibrosis Lung Environment

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

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102 Development of Biosensor Chip for Detection of Specific Antibodies to HSV-1

Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

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101 Information and Communication Technology Skills of Finnish Students in Particular by Gender

Authors: Antero J. S. Kivinen, Suvi-Sadetta Kaarakainen

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Digitalization touches every aspect of contemporary society, changing the way we live our everyday life. Contemporary society is sometimes described as knowledge society including unprecedented amount of information people face daily. The tools to manage this information flow are ICT-skills which are both technical skills and reflective skills needed to manage incoming information. Therefore schools are under constant pressure of revision. In the latest Programme for International Student Assessment (PISA) girls have been outperforming boys in all Organization for Economic Co-operation and Development (OECD) member countries and the gender gap between girls and boys is widest in Finland. This paper presents results of the Comprehensive Schools in the Digital Age project of RUSE, University of Turku. The project is in connection with Finnish Government Analysis, Assessment and Research Activities. First of all, this paper examines gender differences in ICT-skills of Finnish upper comprehensive school students. Secondly, it explores in which way differences are changing when students proceed to upper secondary and vocational education. ICT skills are measured using a performance-based ICT-skill test. Data is collected in 3 phases, January-March 2017 (upper comprehensive schools, n=5455), September-December 2017 (upper secondary and vocational schools, n~3500) and January-March 2018 (Upper comprehensive schools). The age of upper comprehensive school student’s is 15-16 and upper secondary and vocational school 16-18. The test is divided into 6 categories: basic operations, productivity software, social networking and communication, content creation and publishing, applications and requirements for the ICT study programs. Students have filled a survey about their ICT-usage and study materials they use in school and home. Cronbach's alpha was used to estimate the reliability of the ICT skill test. Statistical differences between genders were examined using two-tailed independent samples t-test. Results of first data from upper comprehensive schools show that there is no statistically significant difference in ICT-skill tests total scores between genders (boys 10.24 and girls 10.64, maximum being 36). Although, there were no gender difference in total test scores, there are differences in above mentioned six categories. Girls get better scores on school related and social networking test subjects while boys perform better on more technical oriented subjects. Test scores on basic operations are quite low for both groups. Perhaps these can partly be explained by the fact that the test was made on computers and majority of students ICT-usage consist of smartphones and tablets. Against this background it is important to analyze further the reasons for these differences. In a context of ongoing digitalization of everyday life and especially working life, the significant purpose of this analyses is to find answers how to guarantee the adequate ICT skills for all students.

Keywords: basic education, digitalization, gender differences, ICT-skills, upper comprehensive education, upper secondary education, vocational education

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100 Biosocial Determinants of Maternal and Child Health in Northeast India: A Case Study

Authors: Benrithung Murry

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This paper highlights the biosocial determinants of health-seeking behavior in tribal population groups of northeast India, focusing on maternal and child health. The northeastern region of India is a conglomeration of several ethnic groups, most of which are scheduled as tribal groups. A total of 750 ever-married women in reproductive ages (15-49 years) were interviewed from three tribal groups of Nagaland, India using pre-tested and modified maternal health schedule. Data pertaining to reproductive performance of the mothers and their children health status were collected from 12 villages of Dimapur district, Nagaland, India. The sample for study comprises 212 Angami women, 267 Ao women, and 271 Sumi women, all of which belonging to tribal populations of Northeast India. Sex ratios of 15-49 years in these three populations are 1018.18, 1086.69, and 1106.92, respectively. 90% of the populations in the study are nuclear families, with about 10% of households falling below the poverty line as per the cutoffs for India. Female literacy level in these population groups is higher than the national average of 65.46%; however, about 30% of all married women are not engaged in any sort of earnings. Total fertility rates of these populations are alarming (Total Fertility Rate ≥ 6) and far from replacement fertility level, while infant mortality rates are found to be much lower than the national average of 34 per 1000. The perception and practice of maternal health in this region is unimpressive despite the availability of medical amenities. Only 3 % of mothers in the study have reported 4 times antenatal checkups during last two pregnancies. Other mothers have reported 1 to 3 times of antenatal checkups, but about 25% of them never visited a doctor during the entire pregnancy period. About 15% of mothers never took tetanus injection, while 40% of mothers never took iron folic supplements during pregnancy. Almost half of all women and their husbands do not use birth control measures even for the spacing of children, which has an immense impact on prenatal mortality mainly due to deliberate abortions: the percentage of prenatal mortality among Angami, Ao and Sumi populations is 44.88, 31.88 and 54.98, respectively per 1000 live births. The steep decline in fertility levels in most countries is a consequence of the increasing use of modern methods of contraception. However, among users of birth control measures in these populations, it is seen that most couples use it only after they have the desired number of children, thus its use having no substantial influence in reducing fertility. It is also seen that the majority of the children were only partially vaccinated. With many child deliveries being done at home, many newborns are not administered with polio at birth. Two-third of all children do not have complete basic immunization against polio, diphtheria, tetanus, pertussis, bacillus, and hepatitis besides others. Certain adherence to traditional beliefs and customs apart from the socio-economic factors is believed to have been operating in these populations, which determines their health-seeking behavior. While a more in-depth study combining biological, socio-cultural, economic, and genetic factors is suggested, there is an urgent need for intervention in these populations to combat with the poor maternal and child health status.

Keywords: case study, health behavior, mother and child, northeast india

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