Search results for: Tania Vazquez

Authors: Anton Andreev

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With the collapse of the Soviet Union and the collapse of a large part of the socialist regimes, left-wing parties all over the world entered the space of crisis, of problems with ideology, identity, with the definition of its goals and objectives. First of all, we can say that the communist parties actually lost their foundation. In 1992, despite the victory of left-wing forces, a Broad Front in which was the winner in the struggle against dictatorship plunged into a deep crisis, the nature of which is looking for a new platform, a new foundation, new goals. Thus, in the late 20th century, the party has revised theoretical beliefs and positions. Radical communist ideology was moderated to social reformism. Modern leftist movement in Uruguay is a movement of moderate reform. Left forces suggest going through successive changes. Changes in ideology and ideas have influenced to the understanding of foreign policy. After the collapse of the Soviet Union Broad Front has changed the direction of its diplomacy from the orientation to the Soviet state to support the USA policy. Government formed by Broad Front, supported the integration processes in the South America. Uruguay was developing the cooperation in the framework of MERCOSUR and began to create relationship with the new centers of power in world political space. Uruguay in the early 21st century is a country that starts to play important role in the international arena. Elections of 26 October 2014 should answer the question of support of internal policy of a Broad Front, as well as of the support of the diplomatic work of the "Left" governments of the country.

Keywords: Uruguay, broad front, Vazquez, international dimensions

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Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

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The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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Authors: Javier Navarro Garcia, Narciso Vazquez Carretero

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Eurocode 1, part 1-4, wind actions, includes in its article 1.5 the possibility of using numerical calculation methods to obtain information on the loads acting on a building. On the other hand, the analysis using computational fluids dynamics (CFD) in aerospace, aeronautical, and industrial applications is already in widespread use. The application of techniques based on CFD analysis on the building to study its aerodynamic behavior now opens a whole alternative field of possibilities for civil engineering and architecture; optimization of the results with respect to those obtained by applying the regulations, the possibility of obtaining information on pressures, speeds at any point of the model for each moment, the analysis of turbulence and the possibility of modeling any geometry or configuration. The present work compares the results obtained on a building, with respect to its aerodynamic behavior, from a mathematical model based on the analysis by CFD with the results obtained by applying Eurocode1, part1-4, wind actions. It is verified that the results obtained by CFD techniques suppose an optimization of the wind action that acts on the building with respect to the wind action obtained by applying the Eurocode1, part 1-4, wind actions. In order to carry out this verification, a 45m high square base truncated pyramid building has been taken. The mathematical model on CFD, based on finite volumes, has been calculated using the FLUENT commercial computer application using a scale-resolving simulation (SRS) type large eddy simulation (LES) turbulence model for an atmospheric boundary layer wind with turbulent component in the direction of the flow.

Keywords: aerodynamic, CFD, computacional fluids dynamics, computational mechanics

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Authors: Karla D. Vazquez-Valdez, Bradley P. Wynne

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In the last decades manufacturing needs have been changing, leading to the study of manufacturing methods that were underdeveloped, such as incremental forming processes like shear forming. These processes use rotating tools in constant local contact with the workpiece, which is often also rotating, to generate shape. This means much lower loads to forge large parts and no need for expensive special tooling. Potential has already been established by demonstrating manufacture of high-value products, e.g., turbine and satellite parts, with high dimensional accuracy from difficult to manufacture materials. Thus, huge opportunities exist for these processes to replace the current method of manufacture for a range of high value components, e.g., eliminating lengthy machining, reducing material waste and process times; or the manufacture of a complicated shape without the development of expensive tooling. However, little is known about the exact deformation conditions during processing and why certain materials are better than others for shear forming, leading to a lot of trial and error before production. Three alloys were used for this study: Ti-54M, Jethete M154, and IN718. General Microscopy and Electron Backscatter Diffraction (EBSD) were used to measure strains and orientation maps during shear forming. A Design of Experiments (DOE) analysis was also made in order to understand the impact of process parameters in the properties of the final workpieces. Such information was the key to develop a reliable Finite Element Method (FEM) model that closely resembles the deformation paths of this process. Finally, the potential of these three materials to be shear spun was studied using the FEM model and their Forming Limit Diagram (FLD) which led to the development of a rough methodology for testing the shear spinnability of various metals.

Keywords: shear forming, damage, principal strains, forming limit diagram

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Authors: Gaya Bin Noon, Thoko Hanjahanja-Phiri, Laura Xavier Fadrique, Plinio Pelegrini Morita, Hélène Vaillancourt, Jennifer Teague, Tania Donovska

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Active Assisted Living (AAL) refers to systems designed to improve the quality of life, aid in independence, and create healthier lifestyles for care recipients. As the population ages, there is a pressing need for non-intrusive, continuous, adaptable, and reliable health monitoring tools to support aging in place. AAL has great potential to support these efforts with the wide variety of solutions currently available, but insufficient efforts have been made to address concerns arising from the integration of AAL into care. The purpose of this research was to (1) explore the integration of AAL technologies and data into the clinical pathway, and (2) map data access and governance for AAL technology in order to develop standards for use by policy-makers, technology manufacturers, and developers of smart communities for seniors. This was done through four successive research phases: (1) literature search to explore existing work in this area and identify lessons learned; (2) modeling of the continuum of care; (3) adapting a framework for data governance into the AAL context; and (4) interviews with stakeholders to explore the applicability of previous work. Opportunities for standards found in these research phases included a need for greater consistency in language and technology requirements, better role definition regarding who can access and who is responsible for taking action based on the gathered data, and understanding of the privacy-utility tradeoff inherent in using AAL technologies in care settings.

Keywords: active assisted living, aging in place, internet of things, standards

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Authors: Tania Breton, Dorina Simeonov, Shauna MacEachern

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Ontario’s 10 Year Mental Health and Addictions Strategy has included the establishment of 18 Service Collaborative across the province; cross-sector tables in a specific region coming together to explore mental health and addiction system needs and adopting an intervention to address that need. The process is community led and supported by implementation teams from the Centre for Addiction and Mental Health (CAMH), using the framework of implementation science (IS) to enable evidence-based and sustained change. These justice initiatives are focused on the intersection of the justice system and the mental health and addiction systems. In this presentation, we will share the learnings, achievements and challenges of implementing innovative practices to the mental health and addictions needs of Ontarians within the justice system. Specifically, we will focus on the key points across the justice system - from early intervention and trauma-informed, culturally appropriate services to post-sentence support and community reintegration. Our approach to this work involves external implementation support from the CAMH team including coaching, knowledge exchange, evaluation, Aboriginal engagement and health equity expertise. Agencies supported the implementation of tools and processes which changed practice at the local level. These practices are being scaled up across Ontario and community agencies have come together in an unprecedented collaboration and there is a shared vision of the issues overlapping between the mental health, addictions and justice systems. Working with ministry partners has allowed space for innovation and created an environment where better approaches can be nurtured and spread.

Keywords: implementation, innovation, early identification, mental health and addictions, prevention, systems

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Authors: Tania Sharmin

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Energy use, overheating and thermal discomfort in low-income tropical housing remains an under-researched area. This research attempts to explore these aspects in the Loving Community, a housing colony created for former leprosy patients and their families in Ahmedabad in India. The living conditions in these households and working practices of the inhabitants in terms of how the building and its internal and external spaces are used, will be explored through interviews and monitoring which will be based on a household survey and a focus group discussion (FGD). The findings from the study will provide a unique and in-depth account of how the relocation of the affected households to the new, flood-resistant and architecturally-designed buildings may have affected the dwellers’ household routines (health and well-being, comfort, satisfaction and working practices) and overall living conditions compared to those living in poorly-designed, existing low-income housings. The new houses were built under an innovative building project supported by De Montfort University Leicester (DMU)’s Square Mile India project. A comparison of newly-built and existing building typologies will reveal how building design can affect people’s use of space and energy use. The findings will be helpful to design healthier, energy efficient and socially acceptable low-income housing in future, thus addressing United Nation’s sustainable development goals on three aspects: 3 (health and well-being), 7 (energy) and 11 (safe, resilient and sustainable human settlements). This will further facilitate knowledge exchange between policy makers, developers, designers and occupants focused on strategies to increase stakeholders’ participation in the design process.

Keywords: thermal comfort, energy use, low-income housing, tropical climate

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Authors: Tesfay Gidey Bezabeh, Tânia Sofia Oliveira, Josep Crous-Duran, João H. N. Palma

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Ethiopia's economy depends strongly on Coffea arabica production. Coffee, like many other crops, is sensitive to climate change. An urgent development and application of strategies against the negative impacts of climate change on coffee production is important. Agroforestry-based system is one of the strategies that may ensure sustainable coffee production amidst the likelihood of future impacts of climate change. This system involves the combination of trees in buffer extremes, thereby modifying microclimate conditions. This paper assessed coffee production under 1) coffee monoculture and 2) coffee grown using an agroforestry system, under a) current climate and b) two different future climate change scenarios. The study focused on two representative coffee-growing regions of Ethiopia under different soil, climate, and elevation conditions. A process-based growth model (Yield-SAFE) was used to simulate coffee production for a time horizon of 40 years. Climate change scenarios considered were representative concentration pathways (RCP) 4.5 and 8.5. The results revealed that in monoculture systems, the current coffee yields are between 1200-1250 kg ha⁻¹ yr⁻¹, with an expected decrease between 4-38% and 20-60% in scenarios RCP 4.5 and 8.5, respectively. However, in agroforestry systems, the current yields are between 1600-2200 kg ha⁻¹ yr⁻¹; the decrease was lower, ranging between 4-13% and 16-25% in RCP 4.5 and 8.5 scenarios, respectively. From the results, it can be concluded that coffee production under agroforestry systems has a higher level of resilience when facing future climate change and reinforces the idea of using this type of management in the near future for adapting climate change's negative impacts on coffee production.

Keywords: Albizia gummifera, CORDEX, Ethiopia, HADCM3 model, process-based model

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Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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Authors: Shirajul Islam Khan, Muhammad Ashraful Alam Bhuiyan, Syeda Tania Begum

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Introduction: Acne vulgaris is one of the most common dermatologic conditions and affects the vast majority of people at some point during their lifetime, so effective treatment is of major importance. The failure of usual treatment modalities, teratogenic effects with some severe side effects, and resistance to P.Acne by Retinoides have been focusing on new therapeutic options for the treatment of acne. More recently, pulsed dye laser therapy has been reported to reduce acne lesion counts. The negligible morbidity of these treatment modalities and some other benefits of subsequent acne scar management lead this therapy more attractive. Objective: The objective of this study is to assess the efficacy and safety of pulsed dye laser therapy in the treatment of inflammatory acne vulgaris. Materials and Methods: A prospective clinical trial was done in the Department of Dermatology and Venereology, Combined Military Hospital (CMH), Dhaka, to find out the role of pulse dye laser in the treatment of inflammatory acne vulgaris. The study was carried out with 60 patients with mild to moderate acne vulgaris, and those were treated with pulsed dye laser therapy at baseline and after 4, 8, and 12 weeks. Results: Among 60 patients with inflammatory acne, 42(70%) were in the age group of less than 20 years, and 36(60%) were female. Regarding the number of inflammatory lesions, the baseline mean number (± SD) was 12.77 ± 4.01; after 4 weeks of treatment of inflammatory acne by pulsed dye laser was 7.80 ± 4.11; after 8 weeks of treatment, 6.10 ± 4.03 and after 12 weeks of treatment was 4.17 ± 4.02. After 4 weeks of treatment by pulse dye laser, the level of improvement was excellent at 3.3%, good at 10%, fair at 60%, and poor at 26.7%; after 8 weeks of treatment, excellent was 13.3%, good was 46.7%, the fair was 30% and poor 10% and after 12 weeks of treatment, excellent was 56.7%, good 13.3%, fair 23.3% and poor 6.7%. Regarding safety level, out of 60 patients of inflammatory acne vulgaris treated by pulsed dye laser, about 52(86.7%) patients did not observe any side effects. Conclusions: On the basis of the study results, it can be concluded that pulsed-dye laser is highly effective and well tolerated by patients in the treatment of inflammatory acne.

Keywords: pulsed-dye laser, inflammatory acne, acne vulgaris, retinoids

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Authors: Juan C. Reyes, Linnette Rodríguez, Héctor Villanueva, Jorge Vázquez, Ivonne Rivera

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On July 2020, a private non-profit community health center (HealthProMed) that serves people without a medical insurance plan or with limited resources in one of the most populated areas in San Juan, Puerto Rico, implemented a COVID-19 case investigation and contact-tracing surveillance system. Nursing personnel at the health center completed a computerized case investigation form that was translated, adapted, and modified from CDC’s Patient Under Investigation (PUI) Form. Between July 13, 2020, and March 17, 2022, a total of 9,233 SARS-CoV-2 tests were conducted at the health center, 16.9% of which were classified as confirmed cases (positive molecular test) and 27.7% as probable cases (positive serologic test). Most of the confirmed cases were females (60.0%), under 20 years old (29.1%), and living in their homes (59.1%). In the 14 days before the onset of symptoms, 26.3% of confirmed cases reported going to the supermarket, 22.4% had contact with a known COVID-19 case, and 20.7% went to work. The symptoms most commonly reported were sore throat (33.4%), runny nose (33.3%), cough (24.9%), and headache (23.2%). The most common preexisting medical conditions among confirmed cases were hypertension (19.3%), chronic lung disease including asthma, emphysema, COPD (13.3%), and diabetes mellitus (12.8). Multiple logistic regression analysis revealed that patients who used alcohol frequently during the last two weeks (OR=1.43; 95%CI: 1.15-1.77), those who were in contact with a positive case (OR=1.58; 95%CI: 1.33-1.88) and those who were obese (OR=1.82; 95%CI: 1.24-2.69) were significantly more likely to be a confirmed case after controlling for sociodemographic variables. Implementing a case investigation and contact-tracing component at community health centers can be of great value in the prevention and control of COVID-19 at the community level and could be used in future outbreaks.

Keywords: community health center, Puerto Rico, risk factors, SARS-CoV-2

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Authors: C. Vázquez, L. M. Gigirey, C. P. del Oro, S. Seoane

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Early detection of visual problems plays a key role in the aging process. However, although vision problems are common among older people, the percentage of aging people who perform regular optometric exams is low. In fact, uncorrected refractive errors are one of the main causes of visual impairment in this group of the population. Purpose: To evaluate functional vision of older residents in order to show the urgent need of visual screening programs in Galician nursing homes. Methodology: We examined 364 older adults aged 65 years and over. To measure vision of the daily living, we tested distance and near presenting visual acuity (binocular visual acuity with habitual correction if warn, directional E-Snellen) Presenting near vision was tested at the usual working distance. We defined visual impairment (distance and near) as a presenting visual acuity less than 0.3. Exclusion criteria included immobilized residents unable to reach the USC Dual Sensory Loss Unit for visual screening. Association between categorical variables was performed using chi-square tests. We used Pearson and Spearman correlation tests and the variance analysis to determine differences between groups of interest. Results: 23,1% of participants have visual impairment for distance vision and 16,4% for near vision. The percentage of residents with far and near visual impairment reaches 8,2%. As expected, prevalence of visual impairment increases with age. No differences exist with regard to the level of functional vision between gender. Differences exist between age group respect to distance vision, but not in case of near vision. Conclusion: prevalence of visual impairment is high among the older people tested in this pilot study. This means a high percentage of older people with limitations in their daily life activities. It is necessary to develop an effective vision screening program for early detection of vision problems in Galician nursing homes.

Keywords: functional vision, elders, aging, nursing homes

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Authors: C. Vázquez, L. M. Gigirey, C. P. del Oro, S. Seoane

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Poor vision is common among older people, and several studies show connections between visual impairment and cognitive function. 15 older adult live in Galician Government nursing homes, and cognitive decline is one of the main reasons of admission. Objectives: (1) To evaluate functional far and near vision of older people with cognitive impairment. (2) To determine connections between visual and cognitive state of “our” residents. Methodology: A total of 364 older adults (aged 65 years or more) underwent a visual and cognitive screening. We tested presenting visual acuity (binocular visual acuity with habitual correction if warn) for distance and near vision (E-Snellen, usual working distance for near vision). Binocular presenting visual acuity less than 0.3 was used as cut point for diagnosis of visual impairment. Exclusion criteria included immobilized residents unable to reach the USC Dual Sensory Loss Unit for visual screening. To screen cognition we employed the mini-mental examination test (Spanish version). Analysis of categorical variables was performed using chi-square tests. We utilized Pearson and Spearman correlation tests and the variance analysis to determine differences between groups of interest (SPSS 19.0 version). Results: the percentage of residents with cognitive decline reaches 32.2% Prevalence of visual impairment for distance and near vision increases among those subjects with cognitive impairment respect those with normal cognition. Shift correlation exists between distance visual acuity and mini-mental test (age and sex controlled), and moderate association was found in case of near vision (p<0.01). Conclusion: First results shows that people with cognitive impairment have poor functional distance and near vision than those with normal cognition. Next step will be to analyse the individual contribution of distance and near vision loss on cognition.

Keywords: visual impairment, cognition, aging, nursing homes

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Authors: Muñiz-Lino Marcos Agustín, Vazquez Borbolla Jessica, Licéaga-Escalera Carlos

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The ectomesenchymal tissues involved in tooth development and their remnants are the origin of different odontogenic lesions, including tumors and cysts of the jaws, with a wide range of clinical behaviors. Dentigerous cyst (DC) represents approximately 20% of all cases of odontogenic cysts, and it has been demonstrated that it can develop benign and malignant odontogenic tumors. DC is characterized by bone destruction of the area surrounding the crown of a tooth which has not erupted and it contain is liquid. The treatment of odontogenic tumors and cysts usually are partial or total removal of the jaw, causing important secondary co-morbidities. However, molecules implicated in DC pathogenesis as well in its development to odontogenic tumors remains unknown. A cellular model may be useful to study these molecules, but that model has not been established yet. Here, we reported the establishment of a cell culture derived from a dentigerous cyst. This cell line was named DeCy-1. In spite of its ectomesenchymal morphology, DeCy-1 cells express epithelial markers such as cytokeratins 5, 6, and 8. Furthermore, these cells express the ODAM protein, which is present in odontogenesis and in dental follicle, indicating that DeCy-1 cells derived from odontogenic epithelium. Analysis by electron microscopy of this cell line showed that it has a high vesicular activity, suggesting that DeCy-1 could secrete molecules that may be involved in DC pathogenesis. Thus, secreted proteins were analyzed by PAGE-SDS, where we observed approximately 11 bands. In addition, the capacity of these secretions to degrade proteins was analyzed by gelatin substrate zymography. A degradation band of about 62 kDa was found in these assays. Western blot assays suggested that the matrix metalloproteinase 2 (MMP-2) is responsible of this protease activity. Thus, our results indicate that the establishment of a cell line derived from DC is a useful in vitro model to study the biology of this odontogenic lesion and its participation in the development of odontogenic tumors.

Keywords: dentigerous cyst, MMP20, cancer, cell culture

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Authors: Seema Dutt Bandhu, Yashi Bansal, Tania Moudgil, Barinder Kaur

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Introduction: Scleral perforation during the passage of suture needle is a known complication of strabismus surgery. The present study was conducted to evolve a safe and easy technique of passing the suture needle through the sclera. A scleral tunnel was created with a 26-guage needle through which the suture needle was passed. The rest of the steps of strabismus surgery were carried out as usual. Material and Methods: After taking clearance from the Institutional Ethics Committee, an interventional study was carried out on twenty patients. The scleral tunnel technique was performed on the patients of strabismus after taking written informed consent. Before passing the suture needle through the sclera during strabismus surgery, a tunnel through approximately half the thickness of the sclera was created with the help of a bent 26-gauge needle. The suture needle was then passed through this tunnel. Rest of the steps of the surgery were carried out in the conventional manner. In a control group of same number of patients, the surgery was performed in the conventional method. Both the groups were followed up for any complications. Ease of passing suture and surgeons’ satisfaction with the technique was noted on a 10-point Likert scale. Results: None of the patients in either group suffered from any complications. Four surgeons participated in the study. The average Likert scale score of the surgeons for satisfaction with the technique was 4.5 on a scale of 5. The score for ease of passage of suture needle was 5 on a score of 5. Discussion: Scleral perforation during passing the sutures through the sclera is a known complication of strabismus surgery. Incidence reported is 7.8% It occurs due to inappropriate engagement of the scleral tissue or passage of the suture needle along a wrong axis during the process of passing the suture needle. The needle track technique eases the passage of passing the suture needle through the sclera as the engagement of the scleral tissue can be done with greater control with a 26-guage needle. The surgeons have reported that they are highly satisfied with the technique and they have reported that the technique eased the passage of the suture needle through the sclera.

Keywords: suture, scleral tunnel, strabismus, scleral perforation

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Authors: Heri Hermansyah, Annisa Kurnia, A. Vania Anisya, Adi Surjosatyo, Yopi Sunarya, Rita Arbianti, Tania Surya Utami

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Lipase is one of biocatalyst which is applied commercially for the process in industries, such as bioenergy, food, and pharmaceutical industry. Nowadays, biocatalysts are preferred in industries because they work in mild condition, high specificity, and reduce energy consumption (high pressure and temperature). But, the usage of lipase for industry scale is limited by economic reason due to the high price of lipase and difficulty of the separation system. Immobilization of lipase is one of the solutions to maintain the activity of lipase and reduce separation system in the process. Therefore, we conduct a study about lipase immobilization with the adsorption-cross linking method using glutaraldehyde because this method produces high enzyme loading and stability. Lipase is immobilized on different kind of resin with the various functional group. Highest enzyme loading (76.69%) was achieved by lipase immobilized on anion macroporous which have anion functional group (OH^‑). However, highest activity (24,69 U/g support) through olive oil emulsion method was achieved by lipase immobilized on anion macroporous-chitosan which have amino (NH₂) and anion (OH^-) functional group. In addition, it also success to produce biodiesel until reach yield 50,6% through interesterification reaction and after 4 cycles stable 63.9% relative with initial yield. While for Aspergillus, niger lipase immobilized on anion macroporous-kitosan have unit activity 22,84 U/g resin and yield biodiesel higher than commercial lipase (69,1%) and after 4 cycles stable reach 70.6% relative from initial yield. This shows that optimum functional group on support for immobilization with adsorption-cross linking is the support that contains amino (NH₂) and anion (OH^-) functional group because they can react with glutaraldehyde and binding with enzyme prevent desorption of lipase from support through binding lipase with a functional group on support.

Keywords: adsorption-cross linking, immobilization, lipase, resin

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Authors: Tania Vasquez Mata, Luis A. Herrera, Cristian Arriaga Canon

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Background: Locally advanced breast cancer of the luminal B phenotype, poses challenges due to its variable response to neoadjuvant chemotherapy. A predictive biomarker is needed to identify patients who will not respond to treatment, allowing for alternative therapies. This study aims to validate the use of the lncRNA GATA3-AS1, as a predictive biomarker using RNA in situ hybridization. Research aim: The aim of this study is to determine if GATA3-AS1 can serve as a biomarker for resistance to neoadjuvant chemotherapy in patients with locally advanced luminal B breast cancer. Methodology: The study utilizes RNA in situ hybridization with predesigned probes for GATA3-AS1 on Formalin-Fixed Paraffin-Embedded tissue sections. The samples underwent pretreatment and protease treatment to enable probe penetration. Chromogenic detection and signal evaluation were performed using specific criteria. Findings: Patients who did not respond to neoadjuvant chemotherapy showed a 3+ score for GATA3-AS1, while those who had a complete response had a 1+ score. Theoretical importance: This study demonstrates the potential clinical utility of GATA3-AS1 as a biomarker for resistance to neoadjuvant chemotherapy. Identifying non-responders early on can help avoid unnecessary treatment and explore alternative therapy options. Data collection and analysis procedures: Tissue samples from patients with locally advanced luminal B breast cancer were collected and processed using RNA in situ hybridization. Signal evaluation was conducted under a microscope, and scoring was based on specific criteria. Questions addressed: Can GATA3-AS1 serve as a predictive biomarker for neoadjuvant chemotherapy response in locally advanced luminal B breast cancer? Conclusion: The lncRNA GATA3-AS1 can be used as a biomarker for resistance to neoadjuvant chemotherapy in patients with locally advanced luminal B breast cancer. Its identification through RNA in situ hybridization of tissue obtained from the initial biopsy can aid in treatment decision-making.

Keywords: biomarkers, breast neoplasms, genetics, neoadjuvant therapy, tumor

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Authors: Adrián D. García-Soto, Alejandro Hernández-Martínez, Jesús G. Valdés-Vázquez, Reyna A. Vizguerra-Alvarez

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Reinforced concrete bridges designed by code are intended to achieve target reliability levels adequate for the geographical environment where the code is applicable. Several methods can be used to estimate such reliability levels. Many of them require the establishment of an explicit limit state function (LSF). When such LSF is not available as a close-form expression, the simulation techniques are often employed. The simulation methods are computing intensive and time consuming. Note that if the reliability of real bridges designed by code is of interest, numerical schemes, the finite element method (FEM) or computational mechanics could be required. In these cases, it can be quite difficult (or impossible) to establish a close-form of the LSF, and the simulation techniques may be necessary to compute reliability levels. To overcome the need for a large number of simulations when no explicit LSF is available, the point estimate method (PEM) could be considered as an alternative. It has the advantage that only the probabilistic moments of the random variables are required. However, in the PEM, fitting of the resulting moments of the LSF to a probability density function (PDF) is needed. In the present study, a very simple alternative which allows the assessment of the reliability levels when no explicit LSF is available and without the need of extensive simulations is employed. The alternative includes the use of the PEM, and its applicability is shown by assessing reliability levels of reinforced concrete bridges in Mexico when a numerical scheme is required. Comparisons with results by using the Monte Carlo simulation (MCS) technique are included. To overcome the problem of approximating the probabilistic moments from the PEM to a PDF, a well-known distribution is employed. The approach mixes the PEM and other classic reliability method (first order reliability method, FORM). The results in the present study are in good agreement whit those computed with the MCS. Therefore, the alternative of mixing the reliability methods is a very valuable option to determine reliability levels when no close form of the LSF is available, or if numerical schemes, the FEM or computational mechanics are employed.

Keywords: structural reliability, reinforced concrete bridges, combined approach, point estimate method, monte carlo simulation

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Authors: Shama Ranjan Barua, Tofazzal M. Rakib, Mohammad Alamgir Hossain, Tania Ferdushy, Sharmin Chowdhury

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Rotavirus is one of the main etiologies of neonatal diarrhea in bovine calves that causes significant economic loss in Bangladesh. The present study was carried out to investigate the pathology of neonatal enteritis in calves due to bovine rotavirus infection in south-eastern part of Bangladesh. Rotavirus was identified by using ELISA, RT-PCR (Reverse Transcription Polymerase Chain Reaction), real-time RT-PCR. We examined 12 dead calves with history of diarrhea during necropsy. Among 12 dead calves, in gross examination, 6 were found with pathological changes in intestine, 5 calves had congestion of small intestine and rest one had no distinct pathological changes. Intestinal contents and/or faecal samples of all dead calves were collected and examined to confirm the presence of bovine rotavirus A using Enzyme linked immunosorbant assay (ELISA), RT-PCR and real-time RT-PCR. Out 12 samples, 5 (42%) samples revealed presence of bovine rotavirus A in three diagnostic tests. The histopathological changes were found almost exclusively limited in the small intestine. The lesions of rotaviral enteritis ranged from slight to moderate shortening (atrophy) of villi in the jejunum and ileum with necrotic crypts. The villi were blunt and covered by immature epithelial cells. Infected cells, stained with Haematoxylin and Eosin staining method, showed characteristic syncytia and eosinophilc intracytoplasmic inclusion body. The presence of intracytoplasmic inclusion bodies in enterocytes is the indication of viral etiology. The presence of rotavirus in the affected tissues and/or lesions was confirmed by three different immunological and molecular tests. The findings of histopathological changes will be helpful in future diagnosis of rotaviral infection in dead calves.

Keywords: calves, diarrhea, pathology, rotavirus

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Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel

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In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.

Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids

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Authors: Tania Viju, Bimal P., Naseer M. A.

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This study aims to develop a conceptual framework for the decision support system (DSS), that helps the decision-makers to dynamically choose appropriate safety measures for each identified accident blackspot. An accident blackspot is a segment of road where the frequency of accident occurrence is disproportionately greater than other sections on roadways. According to a report by the World Bank, India accounts for the highest, that is, eleven percent of the global death in road accidents with just one percent of the world’s vehicles. Hence in 2015, the Ministry of Road Transport and Highways of India gave prime importance to the rectification of accident blackspots. To enhance road traffic safety and reduce the traffic accident rate, effectively identifying and rectifying accident blackspots is of great importance. This study helps to understand and evaluate the existing methods in accident blackspot identification and prediction that are used around the world and their application in Indian roadways. The decision support system, with the help of IoT, ICT and smart systems, acts as a management and planning tool for the government for employing efficient and cost-effective rectification strategies. In order to develop a decision criterion, several factors in terms of quantitative as well as qualitative data that influence the safety conditions of the road are analyzed. Factors include past accident severity data, occurrence time, light, weather and road conditions, visibility, driver conditions, junction type, land use, road markings and signs, road geometry, etc. The framework conceptualizes decision-making by classifying blackspot stretches based on factors like accident occurrence time, different climatic and road conditions and suggesting mitigation measures based on these identified factors. The decision support system will help the public administration dynamically manage and plan the necessary safety interventions required to enhance the safety of the road network.

Keywords: decision support system, dynamic management, road accident blackspots, road safety

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Authors: Cesar Augusto Hoil Rosas, José Luis Vázquez Burgos, José Juan Carbajal Hernandez

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White fish (Chirostoma estor estor) is an endemic species that habits in the Patzcuaro Lake, located in Michoacan, Mexico; being an important source of gastronomic and cultural wealth of the area. Actually, it have undergone an immense depopulation of individuals, due to the high fishing, contamination and eutrophication of the lake water, resulting in the possible extinction of this important species. This work proposes a new computational model for monitoring and assessment of critical environmental parameters of the white fish ecosystem. According to an Analytical Hierarchy Process, a mathematical model is built assigning weights to each environmental parameter depending on their water quality importance on the ecosystem. Then, a development of an advanced system for the monitoring, analysis and control of water quality is built using the virtual environment of LabVIEW. As results, we have obtained a global score that indicates the condition level of the water quality in the Chirostoma estor ecosystem (excellent, good, regular and poor), allowing to provide an effective decision making about the environmental parameters that affect the proper culture of the white fish such as temperature, pH and dissolved oxygen. In situ evaluations show regular conditions for a success reproduction and growth rates of this species where the water quality tends to have regular levels. This system emerges as a suitable tool for the water management, where future laws for white fish fishery regulations will result in the reduction of the mortality rate in the early stages of development of the species, which represent the most critical phase. This can guarantees better population sizes than those currently obtained in the aquiculture crop. The main benefit will be seen as a contribution to maintain the cultural and gastronomic wealth of the area and for its inhabitants, since white fish is an important food and economical income of the region, but the species is endangered.

Keywords: Chirostoma estor estor, computational system, lab view, white fish

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Authors: Turki J. Alharbi, Jencia Wong, Dennis Yue, Tania P. Markovic, Julie Hetherington, Ted Wu, Belinda Brooks, Radhika Seimon, Alice Gibson, Stephanie L. Silviera, Amanda Sainsbury, Tanya J. Little

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Fasting has been practiced for centuries and is incorporated into the practices of different religions including Islam, whose followers intermittently fast throughout the month of Ramadan. Thus, Ramadan presents a unique model of prolonged intermittent fasting (IF). Despite a growing body of evidence for a cardio-metabolic and endocrine benefit of IF, detailed studies of the effects of IF on these indices in type 2 diabetes are scarce. We studied 5 subjects with type 2 diabetes (T2DM) and 7 healthy controls (C) at baseline (pre), and in the last week of Ramadan (post). Fasting circulating levels of glucose, HbA1c and lipids, as well as body composition (with DXA) and resting energy expenditure (REE) were measured. Plasma gut hormone levels and appetite responses to a mixed meal were also studied. Data are means±SEM. Ramadan decreased total fat mass (-907±92 g, p=0.001) and trunk fat (-778±190 g, p=0.014) in T2DM but not in controls, without any reductions in lean mass or REE. There was a trend towards a decline in plasma FFA in both groups. Ramadan had no effect on body weight, glycemia, blood pressure, or plasma lipids in either group. In T2DM only, the area under the curve for post-meal plasma ghrelin concentrations increased after Ramadan (pre:6632±1737 vs. post:9025±2518 pg/ml.min-1, p=0.045). Despite this increase in orexigenic ghrelin, subjective appetite scores were not altered by Ramadan. Meal-induced plasma concentrations of the satiety hormone pancreatic polypeptide did not change during Ramadan, but were higher in T2DM compared to controls (post: C: 23486±6677 vs. T2DM: 62193±6880 pg/ml.min-1, p=0.003. In conclusion, Ramadan, as a model for IF appears to have more favourable effects on body composition in T2DM, without adverse effects on metabolic control or subjective appetite. These data suggest that IF may be particularly beneficial in T2DM as a nutritional intervention. Larger studies are warranted.

Keywords: type 2 diabetes, obesity, intermittent fasting, appetite regulating hormones

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Authors: Menna Brown, Tania Domun

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Digital health interventions are effective across a wide array of health conditions spanning physical health, lifestyle behaviour change, and mental health and wellbeing; furthermore, they are rapidly increasing in volume within both the academic literature and society as commercial apps continue to proliferate the digital health market. However, adherence and engagement to digital health interventions remains problematic. Technology-based personalised and tailored reminder strategies can support engagement to digital health interventions. Interventions which support individuals’ mental health and wellbeing are of critical importance in the wake if the COVID-19 pandemic. Student and young person’s mental health has been negatively affected and digital resources continue to offer cost effective means to address wellbeing at a population level. Develop a databank of digital co-produced tailored messages to support engagement to a range of digital health interventions including those focused on mental health and wellbeing, and lifestyle behaviour change. Qualitative research design. Participants discussed their views of health and wellbeing, engagement and adherence to digital health interventions focused around a 12-week wellbeing intervention via a series of focus group discussions. They worked together to co-create content following a participatory design approach. Three focus group discussions were facilitated with (n=15) undergraduate students at one Welsh university to provide an empirically derived, co-produced, databank of (n=145) tailored messages. Messages were explored and categorised thematically, and the following ten themes emerged: Autonomy, Recognition, Guidance, Community, Acceptance, Responsibility, Encouragement, Compassion, Impact and Ease. The findings provide empirically derived, co-produced tailored messages. These have been made available for use, via ‘ACTivate your wellbeing’ a digital, automated, 12-week health and wellbeing intervention programme, based on acceptance and commitment therapy (ACT). The purpose of which is to support future research to evaluate the impact of thematically categorised tailored messages on engagement and adherence to digital health interventions.

Keywords: digital health, engagement, wellbeing, participatory design, positive psychology, co-production

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Authors: Tania Ali Soomro

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In 1839, the British, after the annexation of the port city of Karachi, established a new urban centre consisting of various quarters and introduced new settlements there. These quarters were out of the boundaries of fortified native old area and now contain much of the oldest parts of the city and signify the colonial history of Karachi, in particular the Saddar Bazaar and the neighboring areas of Kharadar and Mithadar. These quarters bestow a mix of functional typology built in a hybrid form of construction - an adaptation of the western architectural attributes to regional requirements and characteristics. This approach is referred to as the Anglo Vernacular, Colonial or the Domestic Gothic architectural form. This research paper investigates the historical and architectural value of one such property: the Empress Market designed by then Municipal Architect, Ar. James Strachan in 1889 as a commemorative monument for the jubilee of Her Majesty the Queen Victoria; Empress of British India, at that time. This paper presents information on the present conservation status of the market building and highlights its role as a catalyst to the community interconnection. This building has survived to present day and functioned well, despite undergoing numerous transformations. A detailed analysis of the bio-degradation (Natural-Chemical dissolution of material) and the bio-deterioration (Manmade-Negative state change of the material) of the building, based on the examination of the prevailing causes of these bio-alterations is carried out, and is presented in form of a damage atlas containing both the categories of bio-alteration/ changes occurred to the building over the time. The research methodology followed in this paper starts with the available archival analysis, physical observation, photographic documentation, the statistics review and the interviews with the direct and indirect stakeholders. The results and findings of this research portray that these bio-alterations and changes are the essential part of the life cycle of Empress Market building which illustrate the historic development of the premise and therefore ought to be given due importance (depending upon their condition) while developing the conservation plan for the building.

Keywords: British colonial architecture, bio-alteration, bio-degradation, bio-deterioration, domestic gothic architectural form

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Authors: Tânia F. C. V. Silva, Eloísa S. S. Vieira, João Pinto da Costa, Rui A. R. Boaventura, Vitor J. P. Vilar

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Sanitary landfill leachates are characterized as a complex mixture of diverse organic and inorganic contaminants, which are usually removed by combining different treatment processes. Due to its simplicity, reliability, high cost-effectiveness and high nitrogen content (mostly under the ammonium form) inherent in this type of effluent, the activated sludge biological process is almost always applied in leachate treatment plants (LTPs). The purpose of this work is to assess the effect of the main nitrification and denitrification variables on the nitrogen's biological removal, from mature leachates. The leachate samples were collected after an aerated lagoon, at a LTP nearby Porto, presenting a high amount of dissolved organic carbon (1.0-1.3 g DOC/L) and ammonium nitrogen (1.1-1.7 g NH4+-N/L). The experiments were carried out in a 1-L lab-scale batch reactor, equipped with a pH, temperature and dissolved oxygen (DO) control system, in order to determine the reaction kinetic constants at unchanging conditions. The nitrification reaction rate was evaluated while varying the (i) operating temperature (15, 20, 25 and 30ºC), (ii) DO concentration interval (0.5-1.0, 1.0-2.0 and 2.0-4.0 mg/L) and (iii) solution pH (not controlled, 7.5-8.5 and 6.5-7.5). At the beginning of most assays, it was verified that the ammonium stripping occurred simultaneously to the nitrification, reaching up to 37% removal of total dissolved nitrogen. The denitrification kinetic constants and the methanol consumptions were calculated for different values of (i) volatile suspended solids (VSS) content (25, 50 and 100 mL of centrifuged sludge in 1 L solution), (ii) pH interval (6.5-7.0, 7.5-8.0 and 8.5-9.0) and (iii) temperature (15, 20, 25 and 30ºC), using effluent previously nitrified. The maximum nitrification rate obtained was 38±2 mg NH4+-N/h/g VSS (25ºC, 0.5-1.0 mg O2/L, pH not controlled), consuming 4.4±0.3 mg CaCO3/mg NH4+-N. The highest denitrification rate achieved was 19±1 mg (NO2--N+NO3--N)/h/g VSS (30ºC, 50 mL of sludge and pH between 7.5 and 8.0), with a C/N consumption ratio of 1.1±0.1 mg CH3OH/mg (NO2--N+NO3--N) and an overall alkalinity production of 3.7±0.3 mg CaCO3/mg (NO2--N+NO3--N). The denitrification process showed to be sensitive to all studied parameters, while the nitrification reaction did not suffered significant change when DO content was changed.

Keywords: mature sanitary landfill leachate, nitrogen removal, nitrification and denitrification parameters, lab-scale activated sludge biological reactor

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Authors: Tania M. Guerrero, Ileana Cortes Santiago

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Estamos Unidos Asylum Project has served more than 2,000 asylum seekers and migrants who are under the Migrant Protection Protocols (MPP) policy in Ciudad Juarez, Mexico. The U.S. policy, implemented in January 2019, has stripped asylum seekers of their rights—forcing people fleeing violence and discrimination to wait in similar or worse conditions from which they fled and navigate their entire asylum process in a different country. Several civil rights groups, including the American Civil Liberties Union (ACLU), challenged MPP in U.S. federal courts in February 2019, arguing a violation of international U.S. obligations towards refugees and asylum-seekers under the 1951 Refugee Convention and the Refugee Act of 1980 in regards to the non-refoulement principle. MPP has influenced Mexico's policies, enforcement, and prioritization of the presence of asylum seekers and migrants; it has also altered the way international non-governmental organizations work at the Mexican Northern border. Estamos Unidos is a project situated in a logistical conundrum, as it provides needed legal services to a population in a legal and humanitarian void, i.e., a liminal space. The liminal space occupied by asylum seekers living under MPP is one that, in today's world, should not be overlooked; it dilutes asylum law and U.S. commitments to international protections. This paper provides analysis of and broader implications from a project whose main goal is to uphold the protections of asylum seekers and international refugee law. The authors identified and analyzed four critical points based on field work conducted since August 2019: (1) strategic coalition building with international, local, and national organizations; (2) brokering between domestic and international contexts and critical legal constraints; (3) flexibility to sudden policy changes and the diverse needs of the multiethnic groups of migrants and asylum seekers served by the project; and (4) the complexity of providing legal assistance to asylum seekers who are survivors of trauma. The authors concur with scholarship when highlighting the erosion of protections of asylum seekers and migrants as a dangerous and unjust global phenomenon.

Keywords: asylum, human rights, migrant protection protocols, refugees law

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Authors: Tania Mannan, Rahelee Zinnat, Fatema Jebunnesa, Israt Ara Hossain

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Background: Exposure to arsenic has known toxic effects but the effect on pregnancy outcomes is not as widely documented especially in women with diabetes. Growing evidence has suggested a potential role of arsenic exposure in the development of gestational diabetes mellitus (GDM). Therefore, we aimed to investigate the association of urinary arsenic (UAs) with birth outcomes in GDM subjects. Methods: Under an observational cross-sectional design a total of 263 GDM subjects (age in years, M±SD, 21±3.7) residing in an arsenic affected area of Bangladesh, were subjected to a 2 sample OGTT at the third trimester of gestation. Among them, 73 GDM and 190 non-GDM subjects enrolled in this study. Clinical and anthropometric measurements were done by standard techniques. Degree of chronic arsenic exposure was assessed by the level of UAs level. According to World Health Organization (WHO) criteria, GDM was diagnosed and neonatal outcomes using APGAR (Activity Pulse Grimace Appearance Respirations) Score, birth weight and size were assessed by a specialist obstetrician. Serum glucose was measured by the Glucose Oxidase method and UAs level was determined by ultraviolet/visible spectrophotometry. Result: Out of the 263 pregnant women, 28% developed GDM. Urinary Arsenic was significantly higher in the GDM as compared to the non-GDM group [UAs, µg/l, M±SD (range), 204.2±67.0 (67.0-377.0) vs 77.3±38.1 (22.0-99.0), p < 0.001]. Activity Pulse Grimace Appearance Respirations Score of the neonates from GDM mothers was significantly lower compared to the neonates from non-GDM mothers [APGAR Score, M±SD, 4.7±0.8 vs. 6.4±0.7, p<0.001]. Pearson’s correlation analysis in GDM subjects revealed that UA levels were found to have a significant positive correlation with both fasting and postprandial serum glucose levels (p < 0.001) and (p < 0.001) respectively. Again, a significant inverse correlation of UAs with birth weight and size was observed (p < 0.001). The APGAR Score of the neonates were found to have a significant negative correlation (p < 0.001) with UAs level. Conclusion: The effect of chronic arsenic exposure is associated with glucose intolerance during pregnancy and it also adversely affects birth outcomes. The study suggests further research on the impact of total arsenic exposure on pregnancy outcomes.

Keywords: APGAR score, arsenic exposure, birth outcome, gestational diabetes mellitus,

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Authors: Rosa Tsegaye Aga, Xuan Jiang, Pavel Vazquez Faci, Siqing Liu, Simon Rayner, Endalkachew Alemu, Markos Abebe

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Tuberculosis (TB) is a major cause of disease globally. In most cases, TB is treatable and curable, but only with the proper treatment. There is a time when drug-resistant TB occurs when bacteria become resistant to the drugs that are used to treat TB. Current strategies to identify drug-resistant TB bacteria are laboratory-based, and it takes a longer time to identify the drug-resistant bacteria and treat the patient accordingly. But machine learning (ML) and data science approaches can offer new approaches to the problem. In this study, we propose to develop an ML-based model to predict the antibiotic resistance phenotypes of TB isolates in minutes and give the right treatment to the patient immediately. The study has been using the whole genome sequence (WGS) of TB isolates as training data that have been extracted from the NCBI repository and contain different countries’ samples to build the ML models. The reason that different countries’ samples have been included is to generalize the large group of TB isolates from different regions in the world. This supports the model to train different behaviors of the TB bacteria and makes the model robust. The model training has been considering three pieces of information that have been extracted from the WGS data to train the model. These are all variants that have been found within the candidate genes (F1), predetermined resistance-associated variants (F2), and only resistance-associated gene information for the particular drug. Two major datasets have been constructed using these three information. F1 and F2 information have been considered as two independent datasets, and the third information is used as a class to label the two datasets. Five machine learning algorithms have been considered to train the model. These are Support Vector Machine (SVM), Random forest (RF), Logistic regression (LR), Gradient Boosting, and Ada boost algorithms. The models have been trained on the datasets F1, F2, and F1F2 that is the F1 and the F2 dataset merged. Additionally, an ensemble approach has been used to train the model. The ensemble approach has been considered to run F1 and F2 datasets on gradient boosting algorithm and use the output as one dataset that is called F1F2 ensemble dataset and train a model using this dataset on the five algorithms. As the experiment shows, the ensemble approach model that has been trained on the Gradient Boosting algorithm outperformed the rest of the models. In conclusion, this study suggests the ensemble approach, that is, the RF + Gradient boosting model, to predict the antibiotic resistance phenotypes of TB isolates by outperforming the rest of the models.

Keywords: machine learning, MTB, WGS, drug resistant TB

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Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

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Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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