Search results for: cellular uptake
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1395

Search results for: cellular uptake

1035 Functional Aspects of Carbonic Anhydrase

Authors: Bashistha Kumar Kanth, Seung Pil Pack

Abstract:

Carbonic anhydrase is ubiquitously distributed in organisms, and is fundamental to many eukaryotic biological processes such as photosynthesis, respiration, CO2 and ion transport, calcification and acid–base balance. However, CA occurs across the spectrum of prokaryotic metabolism in both the archaea and bacteria domains and many individual species contain more than one class. In this review, various roles of CA involved in cellular mechanism are presented to find out the CA functions applicable for industrial use.

Keywords: carbonic anhydrase, mechanism, CO2 sequestration, respiration

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1034 Cost Benefit Analysis: Evaluation among the Millimetre Wavebands and SHF Bands of Small Cell 5G Networks

Authors: Emanuel Teixeira, Anderson Ramos, Marisa Lourenço, Fernando J. Velez, Jon M. Peha

Abstract:

This article discusses the benefit cost analysis aspects of millimetre wavebands (mmWaves) and Super High Frequency (SHF). The devaluation along the distance of the carrier-to-noise-plus-interference ratio with the coverage distance is assessed by considering two different path loss models, the two-slope urban micro Line-of-Sight (UMiLoS) for the SHF band and the modified Friis propagation model, for frequencies above 24 GHz. The equivalent supported throughput is estimated at the 5.62, 28, 38, 60 and 73 GHz frequency bands and the influence of carrier-to-noise-plus-interference ratio in the radio and network optimization process is explored. Mostly owing to the lessening caused by the behaviour of the two-slope propagation model for SHF band, the supported throughput at this band is higher than at the millimetre wavebands only for the longest cell lengths. The benefit cost analysis of these pico-cellular networks was analysed for regular cellular topologies, by considering the unlicensed spectrum. For shortest distances, we can distinguish an optimal of the revenue in percentage terms for values of the cell length, R ≈ 10 m for the millimeter wavebands and for longest distances an optimal of the revenue can be observed at R ≈ 550 m for the 5.62 GHz. It is possible to observe that, for the 5.62 GHz band, the profit is slightly inferior than for millimetre wavebands, for the shortest Rs, and starts to increase for cell lengths approximately equal to the ratio between the break-point distance and the co-channel reuse factor, achieving a maximum for values of R approximately equal to 550 m.

Keywords: millimetre wavebands, SHF band, SINR, cost benefit analysis, 5G

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1033 Community Involvement in Reducing Maternal and Perinatal Mortality in Cross River State, Nigeria: 'The Saving Mother Giving Life' Strategic Approach in Cross River State

Authors: Oluwayemisi Femi-Pius, Kazeem Arogundade, Eberechukwu Eke, Jimmy Eko

Abstract:

Introduction: Globally, community involvement in improving their own health has been widely adopted as a strategy in Sub-Saharan Africa principally to ensure equitable access to essential health care as well as improve the uptake of maternal and newborn health services especially in poor-resource settings. Method: The Saving Mother Giving Life (SMGL) Initiative implemented by Pathfinder International with funding support from USAID conducted a Health Facility Assessment (HFA) and found out that maternal mortality ratio in Cross River State was 812 per 100,000 live birth and perinatal mortality was 160 per 1000 live birth. To reduce maternal and perinatal mortality, Pathfinder International mobilized, selected and trained community members as community volunteers, traditional birth attendants, and emergency transport service volunteer drivers mainly to address the delay in decision making and reaching the health facility among pregnant women. Results: The results showed that maternal mortality ratio in Cross River State decrease by 25% from 812 per 100,000 live birth at baseline to 206 per 100,000 live birth at June 2018 and perinatal mortality reduced by 35% from 160 per 100,000 at baseline to 58 per 1000 live birth at June 2018. Data also show that ANC visit increased from 7,451 to 11,344; institutional delivery increased from 8,931 at baseline to 10,784 in June 2018. There was also a remarkable uptake of post-partum family planning from 0 at baseline to 233 in June 2018. Conclusion: There is clear evidence that community involvement yields positive maternal outcomes and is pivotal for sustaining most health interventions.

Keywords: maternal mortality, Nigeria, pathfinder international, perinatal mortality, saving mother giving life

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1032 The Use of Rule-Based Cellular Automata to Track and Forecast the Dispersal of Classical Biocontrol Agents at Scale, with an Application to the Fopius arisanus Fruit Fly Parasitoid

Authors: Agboka Komi Mensah, John Odindi, Elfatih M. Abdel-Rahman, Onisimo Mutanga, Henri Ez Tonnang

Abstract:

Ecosystems are networks of organisms and populations that form a community of various species interacting within their habitats. Such habitats are defined by abiotic and biotic conditions that establish the initial limits to a population's growth, development, and reproduction. The habitat’s conditions explain the context in which species interact to access resources such as food, water, space, shelter, and mates, allowing for feeding, dispersal, and reproduction. Dispersal is an essential life-history strategy that affects gene flow, resource competition, population dynamics, and species distributions. Despite the importance of dispersal in population dynamics and survival, understanding the mechanism underpinning the dispersal of organisms remains challenging. For instance, when an organism moves into an ecosystem for survival and resource competition, its progression is highly influenced by extrinsic factors such as its physiological state, climatic variables and ability to evade predation. Therefore, greater spatial detail is necessary to understand organism dispersal dynamics. Understanding organisms dispersal can be addressed using empirical and mechanistic modelling approaches, with the adopted approach depending on the study's purpose Cellular automata (CA) is an example of these approaches that have been successfully used in biological studies to analyze the dispersal of living organisms. Cellular automata can be briefly described as occupied cells by an individual that evolves based on proper decisions based on a set of neighbours' rules. However, in the ambit of modelling individual organisms dispersal at the landscape scale, we lack user friendly tools that do not require expertise in mathematical models and computing ability; such as a visual analytics framework for tracking and forecasting the dispersal behaviour of organisms. The term "visual analytics" (VA) describes a semiautomated approach to electronic data processing that is guided by users who can interact with data via an interface. Essentially, VA converts large amounts of quantitative or qualitative data into graphical formats that can be customized based on the operator's needs. Additionally, this approach can be used to enhance the ability of users from various backgrounds to understand data, communicate results, and disseminate information across a wide range of disciplines. To support effective analysis of the dispersal of organisms at the landscape scale, we therefore designed Pydisp which is a free visual data analytics tool for spatiotemporal dispersal modeling built in Python. Its user interface allows users to perform a quick and interactive spatiotemporal analysis of species dispersal using bioecological and climatic data. Pydisp enables reuse and upgrade through the use of simple principles such as Fuzzy cellular automata algorithms. The potential of dispersal modeling is demonstrated in a case study by predicting the dispersal of Fopius arisanus (Sonan), endoparasitoids to control Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) in Kenya. The results obtained from our example clearly illustrate the parasitoid's dispersal process at the landscape level and confirm that dynamic processes in an agroecosystem are better understood when designed using mechanistic modelling approaches. Furthermore, as demonstrated in the example, the built software is highly effective in portraying the dispersal of organisms despite the unavailability of detailed data on the species dispersal mechanisms.

Keywords: cellular automata, fuzzy logic, landscape, spatiotemporal

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1031 Speeding Up Lenia: A Comparative Study Between Existing Implementations and CUDA C++ with OpenGL Interop

Authors: L. Diogo, A. Legrand, J. Nguyen-Cao, J. Rogeau, S. Bornhofen

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Lenia is a system of cellular automata with continuous states, space and time, which surprises not only with the emergence of interesting life-like structures but also with its beauty. This paper reports ongoing research on a GPU implementation of Lenia using CUDA C++ and OpenGL Interoperability. We demonstrate how CUDA as a low-level GPU programming paradigm allows optimizing performance and memory usage of the Lenia algorithm. A comparative analysis through experimental runs with existing implementations shows that the CUDA implementation outperforms the others by one order of magnitude or more. Cellular automata hold significant interest due to their ability to model complex phenomena in systems with simple rules and structures. They allow exploring emergent behavior such as self-organization and adaptation, and find applications in various fields, including computer science, physics, biology, and sociology. Unlike classic cellular automata which rely on discrete cells and values, Lenia generalizes the concept of cellular automata to continuous space, time and states, thus providing additional fluidity and richness in emerging phenomena. In the current literature, there are many implementations of Lenia utilizing various programming languages and visualization libraries. However, each implementation also presents certain drawbacks, which serve as motivation for further research and development. In particular, speed is a critical factor when studying Lenia, for several reasons. Rapid simulation allows researchers to observe the emergence of patterns and behaviors in more configurations, on bigger grids and over longer periods without annoying waiting times. Thereby, they enable the exploration and discovery of new species within the Lenia ecosystem more efficiently. Moreover, faster simulations are beneficial when we include additional time-consuming algorithms such as computer vision or machine learning to evolve and optimize specific Lenia configurations. We developed a Lenia implementation for GPU using the C++ and CUDA programming languages, and CUDA/OpenGL Interoperability for immediate rendering. The goal of our experiment is to benchmark this implementation compared to the existing ones in terms of speed, memory usage, configurability and scalability. In our comparison we focus on the most important Lenia implementations, selected for their prominence, accessibility and widespread use in the scientific community. The implementations include MATLAB, JavaScript, ShaderToy GLSL, Jupyter, Rust and R. The list is not exhaustive but provides a broad view of the principal current approaches and their respective strengths and weaknesses. Our comparison primarily considers computational performance and memory efficiency, as these factors are critical for large-scale simulations, but we also investigate the ease of use and configurability. The experimental runs conducted so far demonstrate that the CUDA C++ implementation outperforms the other implementations by one order of magnitude or more. The benefits of using the GPU become apparent especially with larger grids and convolution kernels. However, our research is still ongoing. We are currently exploring the impact of several software design choices and optimization techniques, such as convolution with Fast Fourier Transforms (FFT), various GPU memory management scenarios, and the trade-off between speed and accuracy using single versus double precision floating point arithmetic. The results will give valuable insights into the practice of parallel programming of the Lenia algorithm, and all conclusions will be thoroughly presented in the conference paper. The final version of our CUDA C++ implementation will be published on github and made freely accessible to the Alife community for further development.

Keywords: artificial life, cellular automaton, GPU optimization, Lenia, comparative analysis.

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1030 Control of Oil Content of Fried Zucchini Slices by Partial Predrying and Process Optimization

Authors: E. Karacabey, Ş. G. Özçelik, M. S. Turan, C. Baltacıoğlu, E. Küçüköner

Abstract:

Main concern about deep-fat-fried food materials is their high final oil contents absorbed during frying process and/or after cooling period, since diet including high content of oil is accepted unhealthy by consumers. Different methods have been evaluated to decrease oil content of fried food stuffs. One promising method is partially drying of food material before frying. In the present study it was aimed to control and decrease the final oil content of zucchini slices by means of partial drying and to optimize process conditions. Conventional oven drying was used to decrease moisture content of zucchini slices at a certain extent. Process performance in terms of oil uptake was evaluated by comparing oil content of predried and then fried zucchini slices with those determined for directly fried ones. For predrying and frying processes, oven temperature and weight loss and frying oil temperature and time pairs were controlled variables, respectively. Zucchini slices were also directly fried for sensory evaluations revealing preferred properties of final product in terms of surface color, moisture content, texture and taste. These properties of directly fried zucchini slices taking the highest score at the end of sensory evaluation were determined and used as targets in optimization procedure. Response surface methodology was used for process optimization. The properties, determined after sensory evaluation, were selected as targets; meanwhile oil content was aimed to be minimized. Results indicated that final oil content of zucchini slices could be reduced from 58% to 46% by controlling conditions of predrying and frying processes. As a result, it was suggested that predrying could be one choose to reduce oil content of fried zucchini slices for health diet. This project (113R015) has been supported by TUBITAK.

Keywords: health process, optimization, response surface methodology, oil uptake, conventional oven

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1029 How Participatory Climate Information Services Assist Farmers to Uptake Rice Disease Forecasts and Manage Diseases in Advance: Evidence from Coastal Bangladesh

Authors: Moriom Akter Mousumi, Spyridon Paparrizos, Fulco Ludwig

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Rice yield reduction due to climate change-induced disease occurrence is becoming a great concern for coastal farmers of Bangladesh. The development of participatory climate information services (CIS) based on farmers’ needs could implicitly facilitate farmers to get disease forecasts and make better decisions to manage diseases. Therefore, this study aimed to investigate how participatory climate information services assist coastal rice farmers to take up rice disease forecasts and better manage rice diseases by improving their informed decision-making. Through participatory approaches, we developed a tailor-made agrometeorological service through the DROP app to forecast rice diseases and manage them in advance. During farmers field schools (FFS) we communicated 7-day disease forecasts during face-to-face weekly meetings using printed paper and, messenger app derived from DROP app. Results show that the majority of the farmers understand disease forecasts through visualization, symbols, and text. The majority of them use disease forecast information directly from the DROP app followed by face-to-face meetings, messenger app, and printed paper. Farmers participation and engagement during capacity building training at FFS also assist them in making more informed decisions and improved management of diseases using both preventive measures and chemical measures throughout the rice cultivation period. We conclude that the development of participatory CIS and the associated capacity-building and training of farmers has increased farmers' understanding and uptake of disease forecasts to better manage of rice diseases. Participatory services such as the DROP app offer great potential as an adaptation option for climate-smart rice production under changing climatic conditions.

Keywords: participatory climate service, disease forecast, disease management, informed decision making, coastal Bangladesg

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1028 Identifying and Optimizing the Critical Excipients in Moisture Activated Dry Granulation Process for Two Anti TB Drugs of Different Aqueous Solubilities

Authors: K. Srujana, Vinay U. Rao, M. Sudhakar

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Isoniazide (INH) a freely water soluble and pyrazinamide (Z) a practically water insoluble first line anti tubercular (TB) drugs were identified as candidates for optimizing the Moisture Activated Dry Granulation (MADG) process. The work focuses on identifying the effect of binder type and concentration as well as the effect of magnesium stearate level on critical quality attributes of Disintegration time (DT) and in vitro dissolution test when the tablets are processed by the MADG process. Also, the level of the drug concentration, binder concentration and fluid addition during the agglomeration stage of the MADG process was evaluated and optimized. For INH, it was identified that for tablets with HPMC as binder at both 2% w/w and 5% w/w level and Magnesium stearate upto 1%w/w as lubrication the DT is within 1 minute and the dissolution rate is the fastest (> 80% in 15 minutes) as compared to when PVP or pregelatinized starch is used as binder. Regarding the process, fast disintegrating and rapidly dissolving tablets are obtained when the level of drug, binder and fluid uptake in agglomeration stage is 25% w/w 0% w/w binder and 0.033%. w/w. At the other 2 levels of these three ingredients, the DT is significantly impacted and dissolution is also slower. For pyrazinamide,it was identified that for the tablets with 2% w/w level of each of PVP as binder and Cross Caramellose Sodium disintegrant the DT is within 2 minutes and the dissolution rate is the fastest(>80 in 15 minutes)as compared to when HPMC or pregelatinized starch is used as binder. This may be attributed to the fact that PVP may be acting as a solubilizer for the practically insoluble Pyrazinamide. Regarding the process,fast dispersing and rapidly disintegrating tablets are obtained when the level of drug, binder and fluid uptake in agglomeration stage is 10% w/w,25% w/w binder and 1% w/w.At the other 2 levels of these three ingredients, the DT is significantly impacted and dissolution is comparatively slower and less complete.

Keywords: agglomeration stage, isoniazide, MADG, moisture distribution stage, pyrazinamide

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1027 Boosting the Agrophysiological Performance of Chickpea Crop (Cicer Arietinum L.) Under Low-P Soil Conditions with the Co-application of Bacterial Consortium (Phosphate Solubilizing Bacteria and Rhizobium) and P-Fertilizers (RP and TSP Forms)

Authors: Rym Saidi, Pape Alioune Ndiaye, Ibnyasser Ammar, Zineb Rchiad, Khalid Daoui, Issam Kadmiri Meftahi, Adnane Bargaz

Abstract:

Chickpea (Cicer arietinum L.) is an important leguminous crop grown worldwide and plays a significant role in humans’ dietary consumption. Alongside nitrogen (N), low phosphorus (P) availability within agricultural soils is one of the major factors limiting chickpea growth and productivity. The combined application of beneficial bacterial inoculants and Rock P-fertilizer could boost chickpea performance and productivity, increasing P-utilization efficiency and minimizing nutrient losses under P-deficiency conditions. A greenhouse experiment was conducted to evaluate the response of chickpeas to two P-fertilizer forms (RP and TSP) under N2-fixer and P-solubilizer consortium inoculation to improve biological N fixation and P nutrition under P-deficient conditions. Under inoculation, chickpea chlorophyll content and chlorophyll fluorescence (RP+I and TSP+I) were increased compared to uninoculated treatments. The RP+I treatment increased both shoot and root dry weights by 48,80% and 72,68%, respectively, compared to the uninoculated RP fertilized control. Indeed, the bacterial consortium contributed to enhancing root morphological traits (e.g., root volume, surface area, and diameter) of all inoculated treatments versus the uninoculated treatments. Furthermore, soil available P and root inorganic P were significantly improved in RP+I by 162,84% and 73,24%, respectively, compared to uninoculated RP control. Our research outcomes suggest that the co-inoculation of chickpeas with N2-fixing, and P-solubilizing bacteria improves biomass yield and nutrient uptake. Eventually, enhancing chickpea agrophysiological performance, especially in restricted P-availability conditions.

Keywords: chickpea, consortium, beneficial bacterial inoculants, phosphorus deficiency, rock p-fertilizer, nutrient uptake

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1026 Enhancement of Radiosensitization by Aptamer 5TR1-Functionalized AgNCs for Triple-Negative Breast Cancer

Authors: Xuechun Kan, Dongdong Li, Fan Li, Peidang Liu

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Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer with a poor prognosis, and radiotherapy is one of the main treatment methods. However, due to the obvious resistance of tumor cells to radiotherapy, high dose of ionizing radiation is required during radiotherapy, which causes serious damage to normal tissues near the tumor. Therefore, how to improve radiotherapy resistance and enhance the specific killing of tumor cells by radiation is a hot issue that needs to be solved in clinic. Recent studies have shown that silver-based nanoparticles have strong radiosensitization, and silver nanoclusters (AgNCs) also provide a broad prospect for tumor targeted radiosensitization therapy due to their ultra-small size, low toxicity or non-toxicity, self-fluorescence and strong photostability. Aptamer 5TR1 is a 25-base oligonucleotide aptamer that can specifically bind to mucin-1 highly expressed on the membrane surface of TNBC 4T1 cells, and can be used as a highly efficient tumor targeting molecule. In this study, AgNCs were synthesized by DNA template based on 5TR1 aptamer (NC-T5-5TR1), and its role as a targeted radiosensitizer in TNBC radiotherapy was investigated. The optimal DNA template was first screened by fluorescence emission spectroscopy, and NC-T5-5TR1 was prepared. NC-T5-5TR1 was characterized by transmission electron microscopy, ultraviolet-visible spectroscopy and dynamic light scattering. The inhibitory effect of NC-T5-5TR1 on cell activity was evaluated using the MTT method. Laser confocal microscopy was employed to observe NC-T5-5TR1 targeting 4T1 cells and verify its self-fluorescence characteristics. The uptake of NC-T5-5TR1 by 4T1 cells was observed by dark-field imaging, and the uptake peak was evaluated by inductively coupled plasma mass spectrometry. The radiation sensitization effect of NC-T5-5TR1 was evaluated through cell cloning and in vivo anti-tumor experiments. Annexin V-FITC/PI double staining flow cytometry was utilized to detect the impact of nanomaterials combined with radiotherapy on apoptosis. The results demonstrated that the particle size of NC-T5-5TR1 is about 2 nm, and the UV-visible absorption spectrum detection verifies the successful construction of NC-T5-5TR1, and it shows good dispersion. NC-T5-5TR1 significantly inhibited the activity of 4T1 cells and effectively targeted and fluoresced within 4T1 cells. The uptake of NC-T5-5TR1 reached its peak at 3 h in the tumor area. Compared with AgNCs without aptamer modification, NC-T5-5TR1 exhibited superior radiation sensitization, and combined radiotherapy significantly inhibited the activity of 4T1 cells and tumor growth in 4T1-bearing mice. The apoptosis level of NC-T5-5TR1 combined with radiation was significantly increased. These findings provide important theoretical and experimental support for NC-T5-5TR1 as a radiation sensitizer for TNBC.

Keywords: 5TR1 aptamer, silver nanoclusters, radio sensitization, triple-negative breast cancer

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1025 In Situ Volume Imaging of Cleared Mice Seminiferous Tubules Opens New Window to Study Spermatogenic Process in 3D

Authors: Lukas Ded

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Studying the tissue structure and histogenesis in the natural, 3D context is challenging but highly beneficial process. Contrary to classical approach of the physical tissue sectioning and subsequent imaging, it enables to study the relationships of individual cellular and histological structures in their native context. Recent developments in the tissue clearing approaches and microscopic volume imaging/data processing enable the application of these methods also in the areas of developmental and reproductive biology. Here, using the CLARITY tissue procedure and 3D confocal volume imaging we optimized the protocol for clearing, staining and imaging of the mice seminiferous tubules isolated from the testes without cardiac perfusion procedure. Our approach enables the high magnification and fine resolution axial imaging of the whole diameter of the seminiferous tubules with possible unlimited lateral length imaging. Hence, the large continuous pieces of the seminiferous tubule can be scanned and digitally reconstructed for the study of the single tubule seminiferous stages using nuclear dyes. Furthermore, the application of the antibodies and various molecular dyes can be used for molecular labeling of individual cellular and subcellular structures and resulting 3D images can highly increase our understanding of the spatiotemporal aspects of the seminiferous tubules development and sperm ultrastructure formation. Finally, our newly developed algorithms for 3D data processing enable the massive parallel processing of the large amount of individual cell and tissue fluorescent signatures and building the robust spermatogenic models under physiological and pathological conditions.

Keywords: CLARITY, spermatogenesis, testis, tissue clearing, volume imaging

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1024 Improving Quality of Family Planning Services in Pakistan

Authors: Mohammad Zakir, Saamia Shams

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Background: Provision of quality family planning services remarkably contribute towards increased uptake of modern contraceptive methods and have important implications on reducing fertility rates. The quality of care in family planning has beneficial impact on reproductive health of women, yet little empirical evidence is present to show the relationship between the impact of adequate training of Community Mid Wives (CMW) and quality family planning services. Aim: This study aimed to enhance the knowledge and counseling skills of CMWs in improving the access to quality client-centered family planning services in Pakistan. Methodology: A quasi-experimental longitudinal study using Initial Quality Assurance Scores-Training-Post Training Quality Assurance Scores design with a non- equivalent control group was adopted to compare a set of experimental CMWs that received four days training package including Family Planning Methods, Counselling, Communication skills and Practical training on IUCD insertion with a set of comparison CMWs that did not receive any intervention. A sample size of 100 CMW from Suraj Social Franchise (SSF) private providers was recruited from both urban and rural Pakistan. Results: Significant improvement in the family planning knowledge and counseling skills (p< 0.001) of the CMWs was evident in the experimental group as compared to comparison group with p > 0.05. Non- significant association between pre-test level family planning knowledge and counseling skills was observed in both the groups (p>0.05). Conclusion: The findings demonstrate that adequate training is an important determinant of quality of family planning services received by clients. Provider level training increases the likelihood of contraceptives uptake and decreases the likelihood of both unintended and unwanted pregnancies. Enhancing quality of family planning services may significantly help reduce the fertility and improve the reproductive health indicators of women in Pakistan.

Keywords: community mid wives, family planning services, quality of care, training

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1023 Metal Contents in Bird Feathers (Columba livia) from Mt Etna Volcano: Volcanic Plume Contribution and Biological Fractionation

Authors: Edda E. Falcone, Cinzia Federico, Sergio Bellomo, Lorenzo Brusca, Manfredi Longo, Walter D’Alessandro

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Although trace metals are an essential element for living beings, they can become toxic at high concentrations. Their potential toxicity is related not only to the total content in the environment but mostly upon their bioavailability. Volcanoes are important natural metal emitters and they can deeply affect the quality of air, water and soils, as well as the human health. Trace metals tend to accumulate in the tissues of living organisms, depending on the metal contents in food, air and water and on the exposure time. Birds are considered as bioindicators of interest, because their feathers directly reflects the metals uptake from the blood. Birds are exposed to the atmospheric pollution through the contact with rainfall, dust, and aerosol, and they accumulate metals over the whole life cycle. We report on the first data combining the rainfall metal content in three different areas of Mt Etna, variably fumigated by the volcanic plume, and the metal contents in the feathers of pigeons, collected in the same areas. Rainfall samples were collected from three rain gauges placed at different elevation on the Eastern flank of the volcano, the most exposed to airborne plume, filtered, treated with HNO₃ Suprapur-grade and analyzed for Fe, Cr, Co, Ni, Se, Zn, Cu, Sr, Ba, Cd and As by ICP-MS technique, and major ions by ion chromatography. Feathers were collected from single individuals, in the same areas where the rain gauges were installed. Additionally, some samples were collected in an urban area, poorly interested by the volcanic plume. The samples were rinsed in MilliQ water and acetone, dried at 50°C until constant weight and digested in a mixture of 2:1 HNO₃ (65%) - H₂O₂ (30%) Suprapur-grade for 25-50 mg of sample, in a bath at near-to-boiling temperature. The solutions were diluted up to 20 ml prior to be analyzed by ICP-MS. The rainfall samples most contaminated by the plume were collected at close distance from the summit craters (less than 6 km), and show lower pH values and higher concentrations for all analyzed metals relative to those from the sites at lower elevation. Analyzed samples are enriched in both metals directly emitted by the volcanic plume and transported by acidic gases (SO₂, HCl, HF), and metals leached from the airborne volcanic ash. Feathers show different patterns in the different sites related to the exposure to natural or anthropogenic pollutants. They show abundance ratios similar to rainfall for lithophile elements (Ba, Sr), whereas are enriched in Zn and Se, known for their antioxidant properties, probably as adaptive response to oxidative stress induced by toxic metal exposure. The pigeons revealed a clear heterogeneity of metal uptake in the different parts of the volcano, as an effect of volcanic plume impact. Additionally, some physiological processes can modify the fate of some metals after uptake and this offer some insights for translational studies.

Keywords: bioindicators, environmental pollution, feathers, trace metals, volcanic plume

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1022 Microstructural Evolution of Maraging Steels from Powder Particles to Additively Manufactured Samples

Authors: Seyedamirreza Shamsdini, Mohsen Mohammadi

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In this research, 18Ni-300 maraging steel powder particles are investigated by studying particle size distribution along with their morphology and grain structure. The powder analysis shows mostly spherical morphologies with cellular structures. A laser-based additive manufacturing process, selective laser melting (SLM) is used to produce samples for further investigation of mechanical properties and microstructure. Several uniaxial tensile tests are performed on the as-built parts to evaluate the mechanical properties. The macroscopic properties, as well as microscopic studies, are then investigated on the printed parts. Hardness measurements, as well as porosity levels, are measured for each sample and are correlated with microstructures through electron microscopy techniques such as Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The grain structure is studied for the as-printed specimens and compared to the powder particle microstructure. The cellular structure of the printed samples is observed to have dendritic forms with dendrite width dimensions similar to the powder particle cells. The process parameter is changed, and the study is performed for different powder layer thickness, and the resultant mechanical properties and grain structure are shown to be similar. A phase study is conducted both on the powder and the printed samples using X-Ray Diffraction (XRD) techniques, and the austenite phase is observed to at first decrease due to the manufacturing process and again during the uniaxial tensile deformation. The martensitic structure is formed in the first stage based on the heating cycles of the manufacturing process and the remaining austenite is shown to be transformed to martensite due to different deformation mechanisms.

Keywords: additive manufacturing, maraging steel, mechanical properties, microstructure

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1021 Inhalable Lipid-Coated-Chitosan Nano-Embedded Microdroplets of an Antifungal Drug for Deep Lung Delivery

Authors: Ranjot Kaur, Om P. Katare, Anupama Sharma, Sarah R. Dennison, Kamalinder K. Singh, Bhupinder Singh

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Respiratory microbial infections being among the top leading cause of death worldwide are difficult to treat as the microbes reside deep inside the airways, where only a small fraction of drug can access after traditional oral or parenteral routes. As a result, high doses of drugs are required to maintain drug levels above minimum inhibitory concentrations (MIC) at the infection site, unfortunately leading to severe systemic side-effects. Therefore, delivering antimicrobials directly to the respiratory tract provides an attractive way out in such situations. In this context, current study embarks on the systematic development of lung lia pid-modified chitosan nanoparticles for inhalation of voriconazole. Following the principles of quality by design, the chitosan nanoparticles were prepared by ionic gelation method and further coated with major lung lipid by precipitation method. The factor screening studies were performed by fractional factorial design, followed by optimization of the nanoparticles by Box-Behnken Design. The optimized formulation has a particle size range of 170-180nm, PDI 0.3-0.4, zeta potential 14-17, entrapment efficiency 45-50% and drug loading of 3-5%. The presence of a lipid coating was confirmed by FESEM, FTIR, and X-RD. Furthermore, the nanoparticles were found to be safe upto 40µg/ml on A549 and Calu-3 cell lines. The quantitative and qualitative uptake studies also revealed the uptake of nanoparticles in lung epithelial cells. Moreover, the data from Spraytec and next-generation impactor studies confirmed the deposition of nanoparticles in lower airways. Also, the interaction of nanoparticles with DPPC monolayers signifies its biocompatibility with lungs. Overall, the study describes the methodology and potential of lipid-coated chitosan nanoparticles in futuristic inhalation nanomedicine for the management of pulmonary aspergillosis.

Keywords: dipalmitoylphosphatidylcholine, nebulization, DPPC monolayers, quality-by-design

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1020 Hydration Matters: Impact on 3 km Running Performance in Trained Male Athletes Under Heat Conditions

Authors: Zhaoqi He

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Research Context: Endurance performance in hot environments is influenced by the interplay of hydration status and physiological responses. This study aims to investigate how dehydration, up to 2.11% body weight loss, affects the 3 km running performance of trained male athletes under conditions mimicking high temperatures. Methodology: In a randomized crossover design, five male athletes participated in two trials – euhydrated (EU) and dehydrated (HYPO). Both trials included a 70-minute preload run at 55-60% VO2max in 32°C and 50% humidity, followed by a 3-kilometer time trial. Fluid intake was restricted in HYPO to induce a 2.11% body weight loss. Physiological metrics, including heart rate, core temperature, and oxygen uptake, were measured, along with perceptual metrics like perceived exertion and thirst sensation. Findings: The 3-kilometer run completion times showed no significant differences between EU and HYPO trials (p=0.944). Physiological indicators, including heart rate, core temperature, and oxygen uptake, did not significantly vary (p>0.05). Thirst sensation was markedly higher in HYPO (p=0.013), confirming successful induction of dehydration. Other perceptual metrics and gastrointestinal comfort remained consistent. Conclusion: Contrary to the hypothesis, the study reveals that dehydration, inducing up to 2.11% body weight loss, does not significantly impair 3 km running performance in trained male athletes under hot conditions. Thirst sensation was notably higher in the dehydrated state, emphasizing the importance of considering perceptual factors in hydration strategies. The findings suggest that trained runners can maintain performance despite moderate dehydration, highlighting the need for nuanced hydration guidelines in hot-weather running.

Keywords: hypohydration, euhydration, hot environment, 3km running time trial, endurance performance, trained athletes, perceptual metrics, dehydration impact, physiological responses, hydration strategies

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1019 EDTA Assisted Phytoremediation of Cadmium by Enhancing Growth and Antioxidant Defense System in Brassica napus L.

Authors: Mujahid Farid, Shafaqat Ali, Muhammad Bilal Shakoor

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Heavy metals pollution of soil is a prevalent global problem and oilseed rape (Brassica napus L.) are considered useful for the restoration of metal contaminated soils. Phytoextraction is an in-situ environment-friendly technique for the clean-up of contaminated soils. Response to cadmium (Cd) toxicity in combination with a chelator, Ethylenediamminetetraacetic acid (EDTA) was studied in oilseed rape grown hydroponically in greenhouse conditions under three levels of Cd (0, 10, and 50 µM) and two levels of EDTA (0 and 2.5 mM). Cd decreased plant growth, biomass and chlorophyll concentrations while the application of EDTA enhanced plant growth by reducing Cd-induced effects in Cd-stressed plants. Significant decrease in photosynthetic parameters was found by the Cd alone. Addition of EDTA improved the net photosynthetic and gas exchange capacity of plants under Cd stress. Cd at 10 and 50 μM significantly increased electrolyte leakage, the production of hydrogen peroxidase (H2O2) and malondialdehyde (MDA) and a significant reduction was observed in the activities of catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX), and superoxide dismutase under Cd stress plants. Application of EDTA at the rate of 2.5 mM alone and with combination of Cd increased the antioxidant enzymes activities and reduced the electrolyte leakage and production of H2O2 and MDA. Oilseed rape (Brassica napus L.) actively accumulated Cd in roots, stems and leaves and the addition of EDTA boosted the uptake and accumulation of Cd in oil seed rape by dissociating Cd in culture media. The present results suggest that under 8 weeks Cd-induced stress, application of EDTA significantly improve plant growth, chlorophyll content, photosynthetic, gas exchange capacity, improving enzymes activities and increased the metal uptake in roots, stems and leaves of oilseed rape (Brassica napus L.) respectively.

Keywords: antioxidant enzymes, cadmium, chelator, EDTA, growth, oilseed rape

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1018 “Chasing Hope”: Parents’ Perspectives on Complementary and Alternative Interventions for Autism Spectrum Disorder Children in Kazakhstan

Authors: Sofiya An, Akbota Kanderzhanova, Assel Akhmetova, Faye Foster, Chee K. Chan

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Healthcare, education and social support for children with autism in Kazakhstan has been evolving and transforming over the last three decades. There is still limited knowledge of the use of complementary and alternative medicine by families caring for autistic children in this post-Soviet region. An exploratory qualitative focus group study of Kazakhstani families was carried out to capture and understand their experiences of using complementary and alternative (CAM) medicine. A total of six focus groups were conducted in five cities across the country including Nur-Sultan, Almaty, Kyzylorda, Karaganda and Taraz. The perceived factors driving the availability, choice, and use of complementary and alternative medicine by families of autistic children in the country were distilled and evaluated. The data collected was analyzed using a framework analysis and themes and subthemes were developed. Two major themes stood out. The first was the “unmet needs”, which relates to the predisposing factors that motivate parents to CAM uptake, and the second was the “chasing hope”, which relates to the enabling factors that facilitate parents’ uptake of CAM. Fear of missing out (FOMO) is a latent underlying motivation underscoring these two themes as well. Parents of autism spectrum disorder (ASD) children in Kazakhstan have to deal with many challenges when seeking treatment for their children with ASD. They are prepared and resort to try out whatever CAM interventions available. The motivation and rationale of choice of use is driven by the lack of options and the hope of any potential positive outcome rather than from rational decisions based on efficacy or the evidence-based data of CAM. Parents get desperate and are willing to try CAM regardless of and independent of their cultural and belief systems and they do not want to miss out just in case it might work. This study also gives an international and cross-cultural perspective on the motives, choice and practice of parents with ASD children using CAM in Kazakhstan, a Central Asian country.

Keywords: autism spectrum disorder, Central Asia, complementary and alternative medicine, cross-cultural perspective, qualitative research

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1017 Conceptualizing Health-Seeking Behavior among Adolescents and Youth with Substance Use Disorder in Urban Kwazulu-Natal. A Candidacy Framework Analysis

Authors: Siphesihle Hlongwane

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Background: Globally, alcohol consumption, smoking, and the use of illicit drugs kill more than 11.8 million people each year. In sub-Saharan Africa, substance abuse is responsible for more than 6.4% of all deaths recorded and about 4.7% of all Disability Adjusted Life Years (DALYs), with numbers still expected to grow if no drastic measures are taken to curb and address drug use. In a setting where substance use is rife, understanding contextual factors that influence an individual’s perceived eligibility to seek rehabilitation is paramount. Using the candidacy framework, we unpack how situational factors influence an individual’s perceived eligibility for healthcare uptake in adolescents and youth with substance use disorder (SUD). Methods: The candidacy framework is concerned with how people consider their eligibility for accessing a health service. The study collected and analyzed primary qualitative data to answer the research question. Data were collected between January and July 2022 on participants aged between 18 and 35 for drug users and 18 to 60 for family members. Participants include 20 previous and current drug users and 20 family members that experience the effects of addiction. A pre-drafted semi-structured interview guide was administered to a conveniently sampled population supplemented with a referral sampling method. Data were thematically analyzed using the NVivo 12pro software to manage the data. Findings: Our findings show that people with substance use disorders are aware of their drug use habits and acknowledge their candidacy for health services. Candidacy for health services is also acknowledged by those around them, such as family members and peers, and as such, information on the navigation of health services for drug users is shared by those who have attended health services, those affected by drug use, and this includes health service research by family members to identify accessible health services. While participants reported willingness to quit drug use if assistance is provided, the permeability of health care services is hindered by both individual determinations to quit drug use from long-time use and the availability of health services for drug users, such as rehabilitation centers. Our findings also show that drug users are conscious and can articulate their ailments; however, the hunt for the next dose of drugs and long waiting cues for health service acquisition overshadows their claim to health services. Participants reported a mixture of treatments prescribed, with some more gruesome than others prescribed, thus serving as both a facilitator and barrier for health service uptake. Despite some unorthodox forms of treatments prescribed in health care, the majority of those who enter treatment complete the process of treatment, although some are met with setbacks and sometimes relapse after treatment has finished. Conclusion: Drug users are able to ascertain their candidacy for health services; however, individual and environmental characteristics relating to drug use hinder the use of health services. Drug use interventions need to entice health service uptake as a way to improve candidacy for health use.

Keywords: substance use disorder, rehabilitation, drug use, relapse, South Africa, candidacy framework

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1016 Engineering Ligand-Free Biodegradable-Based Nanoparticles for Cell Attachment and Growth

Authors: Simone F. Medeiros, Isabela F. Santos, Rodolfo M. Moraes, Jaspreet K. Kular, Marcus A. Johns, Ram Sharma, Amilton M. Santos

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Tissue engineering aims to develop alternatives to treat damaged tissues by promoting their regeneration. Its basic principle is to place cells on a scaffold capable of promoting cell functions, and for this purpose, polymeric nanoparticles have been successfully used due to the ability of some macro chains to mimic the extracellular matrix and influence cell functions. In general, nanoparticles require surface chemical modification to achieve cell adhesion, and recent advances in their synthesis include methods for modifying the ligand density and distribution onto nanoparticles surface. However, this work reports the development of biodegradable polymeric nanoparticles capable of promoting cellular adhesion without any surface chemical modification by ligands. Biocompatible and biodegradable nanoparticles based on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBHV) were synthesized by solvent evaporation method. The produced nanoparticles were small in size (85 and 125 nm) and colloidally stable against time in aqueous solution. Morphology evaluation showed their spherical shape with small polydispersity. Human osteoblast-like cells (MG63) were cultured in the presence of PHBHV nanoparticles, and growth kinetics were compared to those grown on tissue culture polystyrene (TCPS). Cell attachment on non-tissue culture polystyrene (non-TCPS) pre-coated with nanoparticles was assessed and compared to attachment on TCPS. These findings reveal the potential of PHBHV nanoparticles for cell adhesion and growth, without requiring a matrix ligand to support cells, to be used as scaffolds, in tissue engineering applications.

Keywords: tissue engineering, PHBHV, stem cells, cellular attachment

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1015 Quantum Dot – DNA Conjugates for Biological Applications

Authors: A. Banerjee, C. Grazon, B. Nadal, T. Pons, Y. Krishnan, B. Dubertret

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Quantum Dots (QDs) have emerged as novel fluorescent probes for biomedical applications. The photophysical properties of QDs such as broad absorption, narrow emission spectrum, reduced blinking, and enhanced photostability make them advantageous over organic fluorophores. However, for some biological applications, QDs need to be first targeted to specific intracellular locations. It parallel, base pairing properties and biocompatibility of DNA has been extensively used for biosensing, targetting and intracellular delivery of numerous bioactive agents. The combination of the photophysical properties of QDs and targettability of DNA has yielded fluorescent, stable and targetable nanosensors. QD-DNA conjugates have used in drug delivery, siRNA, intracellular pH sensing and several other applications; and continue to be an active area of research. In this project, a novel method to synthesise QD-DNA conjugates and their applications in bioimaging are investigated. QDs are first solubilized in water using a thiol based amphiphilic co-polymer and, then conjugated to amine functionalized DNA using a heterobifunctional linker. The conjugates are purified by size exclusion chromatography and characterized by UV-Vis absorption and fluorescence spectroscopy, electrophoresis and microscopy. Parameters that influence the conjugation yield such as reducing agents, the excess of salt and pH have been investigated in detail. In optimized reaction conditions, up to 12 single-stranded DNA (15 mer length) can be conjugated per QD. After conjugation, the QDs retain their colloidal stability and high quantum yield; and the DNA is available for hybridization. The reaction has also been successfully tested on QDs emitting different colors and on Gold nanoparticles and therefore highly generalizable. After extensive characterization and robust synthesis of QD-DNA conjugates in vitro, the physical properties of these conjugates in cellular milieu are being invistigated. Modification of QD surface with DNA appears to remarkably alter the fate of QD inside cells and can have potential implications in therapeutic applications.

Keywords: bioimaging, cellular targeting, drug delivery, photostability

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1014 Phi Thickening Induction as a Response to Abiotic Stress in the Orchid Miltoniopsis

Authors: Nurul Aliaa Idris, David A. Collings

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Phi thickenings are specialized secondary cell wall thickenings that are found in the cortex of the roots in a wide range of plant species, including orchids. The role of phi thickenings in the root is still under debate through research have linked environmental conditions, particularly abiotic stresses such as water stress, heavy metal stress and salinity to their induction in the roots. It has also been suggested that phi thickenings may act as a barrier to regulate solute uptake, act as a physical barrier against fungal hyphal penetration due to its resemblance to the Casparian strip and play a mechanical role to support cortical cells. We have investigated phi thickening function in epiphytic orchids of the genus Miltoniopsis through induction experiment against factors such as soil compaction and water stress. The permeability of the phi thickenings in Miltoniopsis was tested through uptake experiments using the fluorescent tracer dyes Calcofluor white, Lucifer yellow and Propidium iodide then viewed with wide-field or confocal microscopy. To test whether phi thickening may prevent fungal colonization in the root cell, fungal re-infection experiment was conducted by inoculating isolated symbiotic fungus to sterile in vitro Miltoniopsis explants. As the movement of fluorescent tracers through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonization of cortical cells, the phi thickenings in Miltoniopsis do not function as a barrier. Phi thickenings were found to be absent in roots grown on agar and remained absent when plants were transplanted to moist soil. However, phi thickenings were induced when plants were transplanted to well-drained media, and by the application of water stress in all soils tested. It is likely that phi thickenings stabilize the root cortex during dehydration. Nevertheless, the varied induction responses present in different plant species suggest that the phi thickenings may play several adaptive roles, instead of just one, depending on species.

Keywords: abiotic stress, Miltoniopsis, orchid, phi thickening

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1013 Raman Spectroscopic of Cardioprotective Mechanism During the Metabolic Inhibition of Heart Cells

Authors: A. Almohammedi, A. J. Hudson, N. M. Storey

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Following ischaemia/reperfusion injury, as in a myocardial infraction, cardiac myocytes undergo oxidative stress which leads to several potential outcomes including; necrotic or apoptotic cell death or dysregulated calcium homeostasis or disruption of the electron transport chain. Several studies have shown that nitric oxide donors protect cardiomyocytes against ischemia and reperfusion. However until present, the mechanism of cardioprotective effect of nitric oxide donor in isolated ventricular cardiomyocytes is not fully understood and has not been investigated before using Raman spectroscopy. For these reasons, the aim of this study was to develop a novel technique, pre-resonance Raman spectroscopy, to investigate the mechanism of cardioprotective effect of nitric oxide donor in isolated ventricular cardiomyocytes exposed to metabolic inhibition and re-energisation. The results demonstrated the first time that Raman microspectroscopy technique has the capability to monitor the metabolic inhibition of cardiomyocytes and to monitor the effectiveness of cardioprotection by nitric oxide donor prior to metabolic inhibition of cardiomyocytes. Metabolic inhibition and reenergisation were used in this study to mimic the low and high oxygen levels experienced by cells during ischaemic and reperfusion treatments. A laser wavelength of 488 nm used in this study has been found to provide the most sensitive means of observe the cellular mechanisms of myoglobin during nitric oxide donor preconditioning, metabolic inhibition and re-energisation and did not cause any damage to the cells. The data also highlight the considerably different cellular responses to metabolic inhibition to ischaemia. Moreover, the data has been shown the relationship between the release of myoglobin and chemical ischemia where that the release of myoglobin from the cell only occurred if a cell did not recover contractility.

Keywords: ex vivo biospectroscopy, Raman spectroscopy, biophotonics, cardiomyocytes, ischaemia / reperfusion injury, cardioprotection, nitric oxide donor

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1012 Expert Opinions about Barriers to Physical Activity among Ghanaian Adults with Type 2 Diabetes Mellitus: A Qualitative Descriptive Study

Authors: Mohammed Amin, Debra Kerr, Yacoba Atiase, Andrea Driscoll

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Background: Physical activity (PA) is a major component of diabetes self-care management; although this is often stressed to patients, many adults with Type 2 Diabetes Mellitus (T2DM)lead sedentary lifestyles, and barriers exist for uptake of PA. Aim: To explore opinions of healthcare professionals about barriers to PA for adults with T2DM in Ghana. Methods: This qualitative descriptive study included 13healthcare professionals (3 Physiotherapists, 3 exercise physiologists, 3 nurses, and 4 physicians) who provide care to individuals with T2DM in Ghana. Data was collected by semi-structured interviews. The social cognitive theory guided the design of the interview schedule. Data was analysed using thematic analysis. Results: Four main themeswere identified: 1) Individual-related factors, 2) Interpersonal factors, 3) Environment-related factors, and 4) Health system-related factors. Fear of injury, existing co-morbidities, and lack of time make it difficult for people with T2DM to engage in PA. Lack of family support, fear of social ridicule, and cultural beliefs prevent uptake of PA. Poorly designed community spaces, including safe walkways and lack of exercise facilities, inhibit PA participation. Few physical therapists and physical therapy centres exist to support PA participation among people with T2DM. Some nurses and doctors lack adequate knowledge to deliver proper PA education to clients, thereby making clients lack the needed support. Conclusion: Adults with T2DM may be restricted from partaking in PA arising from personal and external factors. This study has identified that barriers to exercise need consideration when designing PA programs that aim to improve health outcomes for people with T2DM in Ghana.

Keywords: type 2 diabetes, physical activity, exercise, ghana, qualitative, barriers

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1011 Variation in N₂ Fixation and N Contribution by 30 Groundnut (Arachis hypogaea L.) Varieties Grown in Blesbokfontein Mpumalanga Province, South Africa

Authors: Titus Y. Ngmenzuma, Cherian. Mathews, Feilx D. Dakora

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In Africa, poor nutrient availability, particularly N and P, coupled with low soil moisture due to erratic rainfall, constitutes the major crop production constraints. Although inorganic fertilizers are an option for meeting crop nutrient requirements for increased grain yield, the high cost and scarcity of inorganic inputs make them inaccessible to resource-poor farmers in Africa. Because crops grown on such nutrient-poor soils are micronutrient deficient, incorporating N₂-fixing legumes into cropping systems can sustainably improve crop yield and nutrient accumulation in the grain. In Africa, groundnut can easily form an effective symbiosis with native soil rhizobia, leading to marked N contribution in cropping systems. In this study, field experiments were conducted at Blesbokfontein in Mpumalanga Province to assess N₂ fixation and N contribution by 30 groundnut varieties during the 2018/2019 planting season using the ¹⁵N natural abundance technique. The results revealed marked differences in shoot dry matter yield, symbiotic N contribution, soil N uptake and grain yield among the groundnut varieties. The percent N derived from fixation ranged from 37 to 44% for varieties ICGV131051 and ICGV13984. The amount of N-fixed ranged from 21 to 58 kg/ha for varieties Chinese and IS-07273, soil N uptake from 31 to 80 kg/ha for varieties IS-07947 and IS-07273, and grain yield from 193 to 393 kg/ha for varieties ICGV15033 and ICGV131096, respectively. Compared to earlier studies on groundnut in South Africa, this study has shown low N₂ fixation and N contribution to the cropping systems, possibly due to environmental factors such as low soil moisture. Because the groundnut varieties differed in their growth, symbiotic performance and grain yield, more field testing is required over a range of differing agro-ecologies to identify genotypes suitable for different cropping environments

Keywords: ¹⁵N natural abundance, percent N derived from fixation, amount of N-fixed, grain yield

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1010 Enhancing Mitochondrial Activity and Metabolism in Aging Female Germ Cells: Synergistic Effects of Dual ROCK and ROS Inhibition

Authors: Kuan-Hao Tsui, Li-Te Lin, Chia-Jung Li

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The combination of Y-27632 and Vitamin C significantly enhances the quality of aging germ cells by reducing reactive oxygen species (ROS) production, restoring mitochondrial membrane potential balance, and promoting mitochondrial fusion. The age-related decline in oocyte quality contributes to reduced fertility, increased aneuploidy, and diminished embryo quality, with mitochondrial dysfunction in both oocytes and granulosa cells being a key factor in this decline. Experiments on aging germ cells investigated the effects of the Y-27632 and Vitamin C combination. In vivo studies involved aged mice to assess oocyte maturation and ROS accumulation during culture. The assessment included mitochondrial activity, ROS levels, mitochondrial membrane potential, and mitochondrial dynamics. Cellular energy metabolism and ATP production were also measured. The combination treatment effectively addressed mitochondrial dysfunction and regulated cellular energy metabolism, promoting oxygen respiration and increasing ATP production. In aged mice, this supplement treatment enhanced in vitro oocyte maturation and prevented ROS accumulation in aging oocytes during culture. While these findings are promising, further research is needed to explore the long-term effects and potential side effects of the Y-27632 and Vitamin C combination. Additionally, translating these findings to human subjects requires careful consideration. Overall, the study suggests that the Y-27632 and Vitamin C combination could be a promising intervention to mitigate aging-related dysfunction in germ cells, potentially enhancing oocyte quality, particularly in the context of in vitro fertilization.

Keywords: ovarian aging, supplements, ROS, mitochondria

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1009 PhenoScreen: Development of a Systems Biology Tool for Decision Making in Recurrent Urinary Tract Infections

Authors: Jonathan Josephs-Spaulding, Hannah Rettig, Simon Graspeunter, Jan Rupp, Christoph Kaleta

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Background: Recurrent urinary tract infections (rUTIs) are a global cause of emergency room visits and represent a significant burden for public health systems. Therefore, metatranscriptomic approaches to investigate metabolic exchange and crosstalk between uropathogenic Escherichia coli (UPEC), which is responsible for 90% of UTIs, and collaborating pathogens of the urogenital microbiome is necessary to better understand the pathogenetic processes underlying rUTIs. Objectives: This study aims to determine the level in which uropathogens optimize the host urinary metabolic environment to succeed during invasion. By developing patient-specific metabolic models of infection, these observations can be taken advantage of for the precision treatment of human disease. Methods: To date, we have set up an rUTI patient cohort and observed various urine-associated pathogens. From this cohort, we developed patient-specific metabolic models to predict bladder microbiome metabolism during rUTIs. This was done by creating an in silico metabolomic urine environment, which is representative of human urine. Metabolic models of uptake and cross-feeding of rUTI pathogens were created from genomes in relation to the artificial urine environment. Finally, microbial interactions were constrained by metatranscriptomics to indicate patient-specific metabolic requirements of pathogenic communities. Results: Metabolite uptake and cross-feeding are essential for strain growth; therefore, we plan to design patient-specific treatments by adjusting urinary metabolites through nutritional regimens to counteract uropathogens by depleting essential growth metabolites. These methods will provide mechanistic insights into the metabolic components of rUTI pathogenesis to provide an evidence-based tool for infection treatment.

Keywords: recurrent urinary tract infections, human microbiome, uropathogenic Escherichia coli, UPEC, microbial ecology

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1008 Nanoparticle Induced Neurotoxicity Mediated by Mitochondria

Authors: Nandini Nalika, Suhel Parvez

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Nanotechnology has emerged to play a vital role in developing all through the industrial world with an immense production of nanomaterials including nanoparticles (NPs). Many toxicological studies have confirmed that due to unique small size and physico-chemical properties of NPs (1-100nm), they can be potentially hazardous. Metallic NPs of small size have been shown to induce higher levels of cellular oxidative stress and can easily pass through the Blood Brain Barrier (BBB) and significantly accumulate in brain. With the wide applications of titanium dioxide nanoparticles (TNPs) in day-to-day life in form of cosmetics, paints, sterilisation and so on, there is growing concern regarding the deleterious effects of TNPs on central nervous system and mitochondria appear to be important cellular organelles targeted to the pro-oxidative effects of NPs and an important source that contribute significantly for the production of reactive oxygen species after some toxicity or an injury. The aim of our study was to elucidate the effect of TNPs in anatase form with different concentrations (5-50 µg/ml) following with various oxidative stress markers in isolated brain mitochondria as an in vitro model. Oxidative stress was determined by measuring the different oxidative stress markers like lipid peroxidation as well as the protein carbonyl content which was found to be significantly increased. Reduced glutathione content and major glutathione metabolizing enzymes were also modulated signifying the role of glutathione redox cycle in the pathophysiology of TNPs. The study also includes the mitochondrial enzymes (Complex 1, Complex II, complex IV, Complex V ) and the enzymes showed toxicity in a relatively short time due to the effect of TNPs. The study provide a range of concentration that were toxic to the neuronal cells and data pointing to a general toxicity in brain mitochondria by TNPs, therefore, it is in need to consider the proper utilization of NPs in the environment.

Keywords: mitochondria, nanoparticles, brain, in vitro

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1007 Synthesis, Characterization and Applications of Novel Hydrogels Based On Chitosan Derivatives

Authors: Mahmoud H. Aboul-Ela, Riham R. Mohamed, Magdy W. Sabaa

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Synthesis of cross-linked hydrogels composed of trimethyl chitosan (TMC) and poly(vinyl alcohol) (PVA) in different weight ratios in presence of glutaraldehyde as cross-linking agent. Characterization of the prepared hydrogels was done using FTIR, XRD, SEM and TGA. The prepared hydrogels were investigated as adsorbent materials for some transition metal ions from their aqueous solutions. Moreover, the swell ability of the prepared hydrogels was also investigated in both acidic and alkaline pHs, as well as in simulated body fluid (SBF).

Keywords: trimethyl chitosan, hydrogels, metal uptake, superabsorbent materials

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1006 The Evaluation of Adjuvant Effects of CD154 in a Subunit Vaccine against Classical Swine Fever Virus

Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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