Search results for: gene regulation

Authors: Arushi S. Goyal, Jo Aggarwal, Ravi Jasuja

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Statement of the Problem: Vapes have always been marketed as safer alternatives to traditional cigarettes; however, research suggests that perceived safety of e-cigarettes use may be overstated. While the addictive properties of nicotine have garnered significant scientific interest, the adverse effects of ‘inert’ ingredients in vapes are being investigated only recently. Seemingly harmless components in vapes such as propylene glycol have been shown to damage astrocytes and oligodendrocytes, and certain flavorings are causatively associated with neuroinflammation. With ease of concealment and varied aromas, vape usage amongst high school students continues unabated in countries like India, which have instituted comprehensive bans on e-cigarettes. With overt government ban, there is paucity of public data on determinants of teenage vaping patterns and parental engagement in curbing this debilitating dependency. Additionally, the large body of peer reviewed studies on vaping have been primarily conducted in Western countries. Accordingly, the purpose of this study was to examine the factors affecting the causes and attitudes towards vaping among adolescents in urban India, as well as the gaps in parental awareness. We posit that this study would lay out a reusable framework for extending the studies across conservative societies where adolescents support vaping behavior even with strong governmental policies. Methodology & Theoretical Orientation: Two surveys were used to collect data. Participants from eight private schools in Bangalore completed an online survey. The first survey sampled adolescents aged 14-18, while the second surveyed the parents of children in the same age group from the same schools. Informed consent was obtained from all participants, and all data collected was anonymous. Results: We find substantial discordance in self-reported vape use by the adolescents and the parents’ knowledge of their child’s exposure to vaping. Over one fifth of respondents (22.4%) reported using e-cigarettes, while only 5% of parents reported that their children used e-cigarettes. Even though over 70% of adolescents believe that vaping is addictive, only 22.8% of respondents were aware of the components, or the extent of its impact. While peer pressure is often perceived to be the enabling factor, curiosity was reported as the primary reason for the initiation. Adolescents who vape saw regulations on sales and marketing as the most effective deterrent. In contrast, parents and other students leaned on school infrastructure to intervene. There has been a significant increase in vaping and a substantial discordance between parental perceptions and adolescent vaping. Conclusion & Significance: Despite a complete ban, vapes continue to be easily accessible. The data suggests that an open discussion about the adverse health consequences of untested, “seemingly inert” ingredients in these unregulated vape liquids would galvanize the student community by demystifying vaping. While increased regulation against the sale of vapes deters open use, increased parental involvement could enable open dialog with children and assist in reducing the prevalence of vaping. A reduction in vaping could have a considerable impact on the health and educational outcomes for the youth of India.

Keywords: adolescent, e-cigarettes, health consequences, India, parental awareness, vapes

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Authors: Leigha C. Crout

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This paper will investigate the categorical disregard of the People’s Republic of China (PRC) in establishing and maintaining a baseline standard of civil guarantees for economic migrant women and their dependents. In light of the relative forward strides in terms of policy facilitating the ascension of female workers in China, this oft-invisible subgroup of women remains neglected from the modern-day “iron rice bowl” of the self-identified communist state. This study is being undertaken to rectify the absence of data on this subject and provide a baseline for future studies on the matter, as the human rights of migrants has become an established facet of transnational dialogue and debate. The basic methodology of this research will consist of the evaluation of China’s compliance with its own national guidelines, and the eight international human rights law treaties it has ratified. Data will be extracted and cross-checked from a number of relevant sources to monitor the extent of compliance, including but by no means limited to the United Nations Human Rights Council (UNHRC) Universal Periodic Review (UPR) reports and responses, submissions and responses of international human rights treaty bodies, local and international nongovernmental organizations (NGOs) and their annual reports, and articles and commentaries authored by specialists on the modern state and implementation of Chinese law. Together, these data will illuminate the vast network of compliance that has forced many migrant women to work within situations of extreme economic precarity. The structure will proceed as follows: first, an outline of the current status of migrant workers and the enforcement of stipulated protections will be provided; next, the analysis of the oft-debated regulations directing and the outline of mandatory services guaranteed to external and internal migrants; and finally, a conclusion incorporating various recommendations to improve transparency and gradually decrease the amount of migrant work turned forced labor that typifies the economic migrant experience, especially in the case of women. The internal and international migrant workers in China are bound by different and uncomplimentary systems. The first, which governs Chinese citizens moving to different regions or provinces to find more sustainable employment (internal migrants), is called the hukou (or huji) residency system. This law enforces strict regulation of the movement of peoples, while ensuring that residents of urban areas receive preferential benefits to those received by their so-called “agricultural” resident counterparts. Given the overwhelming presence of the Communist Party of China throughout the vast state, the management of internal migrants and the disregard for foreign domestic workers is, at minimum, a surprising oversight. This paper endeavors to provide a much-needed foundation for future commentary and discussion on the treatment of female migrant workers and their families in the People’s Republic of China.

Keywords: female migrant worker’s rights, the People’s Republic of China, forced labor, Hukou residency system

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Authors: Soma Ghosh, Balaram Mohapatra, Pinaki Sar, Abhijeet Mukherjee

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Microbial role in arsenic (As) mobilization in the groundwater aquifers of Brahmaputra river basin (BRB) in India, severely threatened by high concentrations of As, remains largely unknown. The present study, therefore, is a molecular and ecophysiological characterization of an indigenous bacterium strain IIIJ3-1 isolated from As contaminated groundwater of BRB and application of this strain in several microcosm set ups differing in their organic carbon (OC) source and terminal electron acceptors (TEA), to understand its role in As dissolution under aerobic and anaerobic conditions. Strain IIIJ3-1 was found to be a new facultative anaerobic, gram-positive, endospore-forming strain capable of arsenite (As3+) oxidation and dissimilatory arsenate (As5+) reduction. The bacterium exhibited low genomic (G+C)% content (45 mol%). Although, its 16S rRNA gene sequence revealed a maximum similarity of 99% with Bacillus cereus ATCC 14579(T) but the DNA-DNA relatedness of their genomic DNAs was only 49.9%, which remains well below the value recommended to delimit different species. Abundance of fatty acids iC17:0, iC15:0 and menaquinone (MK) 7 though corroborates its taxonomic affiliation with B. cereus sensu-lato group, presence of hydroxy fatty acids (HFAs), C18:2, MK5 and MK6 marked its uniqueness. Besides being highly As resistant (MTC=10mM As3+, 350mM As5+), metabolically diverse, efficient aerobic As3+ oxidizer; it exhibited near complete dissimilatory reduction of As5+ (1 mM). Utilization of various carbon sources with As5+ as TEA revealed lactate to serve as the best electron donor. Aerobic biotransformation assay yielded a lower Km for As3+ oxidation than As5+ reduction. Arsenic homeostasis was found to be conferred by the presence of arr, arsB, aioB, and acr3(1) genes. Scanning electron microscopy (SEM) coupled with energy dispersive X-ray (EDX) analysis of this bacterium revealed reduction in cell size upon exposure to As and formation of As-rich electron opaque dots following growth with As3+. Incubation of this strain with sediment (sterilised) collected from BRB aquifers under varying OC, TEA and redox conditions revealed that the strain caused highest As mobilization from solid to aqueous phase under anaerobic condition with lactate and nitrate as electron donor and acceptor, respectively. Co-release of highest concentrations of oxalic acid, a well known bioweathering agent, considerable fold increase in viable cell counts and SEM-EDX and X-ray diffraction analysis of the sediment after incubation under this condition indicated that As release is consequent to microbial bioweathering of the minerals. Co-release of other elements statistically proves decoupled release of As with Fe and Zn. Principle component analysis also revealed prominent role of nitrate under aerobic and/or anaerobic condition in As release by strain IIIJ3-1. This study, therefore, is the first to isolate, characterize and reveal As mobilization property of a strain belonging to the Bacillus cereus sensu lato group isolated from highly As contaminated aquifers of Brahmaputra River Basin.

Keywords: anaerobic microcosm, arsenic rich electron opaque dots, Arsenic release, Bacillus strain IIIJ3-1

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Authors: P. J. Sweeney, T. Ahtoniemi, J. Puoliväli, T. Laitinen, K. Lehtimäki, A. Nurmi, D. Wells

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Duchenne muscular dystrophy (DMD) is caused by an X linked mutation in the dystrophin gene; lack of dystrophin causes a progressive muscle necrosis which leads to a progressive decrease in mobility in those suffering from the disease. The MDX mouse, a mutant mouse model which displays a frank dystrophinopathy, is currently widely employed in pre clinical efficacy models for treatments and therapies aimed at DMD. In general the end-points examined within this model have been based on invasive histopathology of muscles and serum biochemical measures like measurement of serum creatine kinase (sCK). It is established that a “critical period” between 4 and 6 weeks exists in the MDX mouse when there is extensive muscle damage that is largely sub clinical but evident with sCK measurements and histopathological staining. However, a full characterization of the MDX model remains largely incomplete especially with respect to the ability to aggravate of the muscle damage beyond the critical period. The purpose of this study was to attempt to aggravate the muscle damage in the MDX mouse and to create a wider, more readily translatable and discernible, therapeutic window for the testing of potential therapies for DMD. The study consisted of subjecting 15 male mutant MDX mice and 15 male wild-type mice to an intense chronic exercise regime that consisted of bi-weekly (two times per week) treadmill sessions over a 12 month period. Each session was 30 minutes in duration and the treadmill speed was gradually built up to 14m/min for the entire session. Baseline plasma creatine kinase (pCK), treadmill training performance and locomotor activity were measured after the “critical period” at around 10 weeks of age and again at 14 weeks of age, 6 months, 9 months and 12 months of age. In addition, kinematic gait analysis was employed using a novel analysis algorithm in order to compare changes in gait and fine motor skills in diseased exercised MDX mice compared to exercised wild type mice and non exercised MDX mice. In addition, a morphological and metabolic profile (including lipid profile), from the muscles most severely affected, the gastrocnemius muscle and the tibialis anterior muscle, was also measured at the same time intervals. Results indicate that by aggravating or exacerbating the underlying muscle damage in the MDX mouse by exercise a more pronounced and severe phenotype in comes to light and this can be picked up earlier by kinematic gait analysis. A reduction in mobility as measured by open field is not apparent at younger ages nor during the critical period, but changes in gait are apparent in the mutant MDX mice. These gait changes coincide with pronounced morphological and metabolic changes by non-invasive anatomical MRI and proton spectroscopy (1H-MRS) we have reported elsewhere. Evidence of a progressive asymmetric pathology in imaging parameters as well as in the kinematic gait analysis was found. Taken together, the data show that chronic exercise regime exacerbates the muscle damage beyond the critical period and the ability to measure through non-invasive means are important factors to consider when performing preclinical efficacy studies in the MDX mouse.

Keywords: Gait, muscular dystrophy, Kinematic analysis, neuromuscular disease

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Authors: Bhargavi G. Iyer, Ojaswi Bhagat

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It is often noted that the value of a novel idea lies in its successful implementation. However, successful implementation requires the nurturing and encouragement of innovation. Nascent competitors are a true representation of innovation in any given industry. A nascent competitor is an entity whose prospective innovation poses a future threat to an incumbent dominant competitor. While a nascent competitor benefits in several ways, it is also exposed significantly and is at greater risk of facing the brunt of exclusionary practises and abusive conduct by dominant incumbent competitors in the industry. This research paper aims to explore the risks and threats faced by nascent competitors and analyse the benefits they accrue as well as the advantages they proffer to the economy; through an analytical, critical study. In such competitive market environments, a rise of the acquisitions of nascent competitors by the incumbent dominants is observed. Therefore, this paper will examine the dynamics of nascent acquisition. Further, this paper hopes to specifically delve into the role of antitrust bodies in regulating nascent acquisition. This paper also aspires to deal with the question how to distinguish harmful from harmless acquisitions in order to facilitate ideal enforcement practice. This paper proposes mechanisms of scrutiny in order to ensure healthy market practises and efficient merger control in the context of nascent acquisitions. Taking into account the scope and nature of the topic, as well as the resources available and accessible, a combination of the methods of doctrinal research and analytical research were employed, utilising secondary sources in order to assess and analyse the subject of research. While legally evaluating the Killer Acquisition theory and the Nascent Potential Acquisition theory, this paper seeks to critically survey the precedents and instances of nascent acquisitions. In addition to affording a compendious account of the legislative framework and regulatory mechanisms in the United States, the United Kingdom, and the European Union; it hopes to suggest an internationally practicable legal foundation for domestic legislation and enforcement to adopt. This paper hopes to appreciate the complexities and uncertainties with respect to nascent acquisitions and attempts to suggest viable and plausible policy measures in antitrust law. It additionally attempts to examine the effects of such nascent acquisitions upon the consumer and the market economy. This paper weighs the argument of shifting the evidentiary burden on to the merging parties in order to improve merger control and regulation and expounds on its discovery of the strengths and weaknesses of the approach. It is posited that an effective combination of factual, legal, and economic analysis of both the acquired and acquiring companies possesses the potential to improve ex post and ex ante merger review outcomes involving nascent companies; thus, preventing anti-competitive practises. This paper concludes with an analysis of the possibility and feasibility of industry-specific identification of anti-competitive nascent acquisitions and implementation of measures accordingly.

Keywords: acquisition, antitrust law, exclusionary practises merger control, nascent competitor

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Authors: Olga Kokoulina

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Today, algorithms are assuming a leading role in various areas of decision-making. Prompted by a promise to provide increased economic efficiency and fuel solutions for pressing societal challenges, algorithmic decision-making is often celebrated as an impartial and constructive substitute for human adjudication. But in the face of this implied objectivity and efficiency, the application of algorithms is also marred with mounting concerns about embedded biases, discrimination, and exclusion. In Europe, vigorous debates on risks and adverse implications of algorithmic decision-making largely revolve around the potential of data protection laws to tackle some of the related issues. For example, one of the often-cited venues to mitigate the impact of potentially unfair decision-making practice is a so-called 'right to explanation'. In essence, the overall right is derived from the provisions of the General Data Protection Regulation (‘GDPR’) ensuring the right of data subjects to access and mandating the obligation of data controllers to provide the relevant information about the existence of automated decision-making and meaningful information about the logic involved. Taking corresponding rights and obligations in the context of the specific provision on automated decision-making in the GDPR, the debates mainly focus on efficacy and the exact scope of the 'right to explanation'. In essence, the underlying logic of the argued remedy lies in a transparency imperative. Allowing data subjects to acquire as much knowledge as possible about the decision-making process means empowering individuals to take control of their data and take action. In other words, forewarned is forearmed. The related discussions and debates are ongoing, comprehensive, and, often, heated. However, they are also frequently misguided and isolated: embracing the data protection law as ultimate and sole lenses are often not sufficient. Mandating the disclosure of technical specifications of employed algorithms in the name of transparency for and empowerment of data subjects potentially encroach on the interests and rights of IPR holders, i.e., business entities behind the algorithms. The study aims at pushing the boundaries of the transparency debate beyond the data protection regime. By systematically analysing legal requirements and current judicial practice, it assesses the limits of the transparency requirement and right to access posed by intellectual property law, namely by copyrights and trade secrets. It is asserted that trade secrets, in particular, present an often-insurmountable obstacle for realising the potential of the transparency requirement. In reaching that conclusion, the study explores the limits of protection afforded by the European Trade Secrets Directive and contrasts them with the scope of respective rights and obligations related to data access and portability enshrined in the GDPR. As shown, the far-reaching scope of the protection under trade secrecy is evidenced both through the assessment of its subject matter as well as through the exceptions from such protection. As a way forward, the study scrutinises several possible legislative solutions, such as flexible interpretation of the public interest exception in trade secrets as well as the introduction of the strict liability regime in case of non-transparent decision-making.

Keywords: algorithms, public interest, trade secrets, transparency

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Authors: Nahla Elsherbiny, Ahmed Ahmed, Hamada Mohammed, Mohamed Ali

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Background: The enterococci may be considered as opportunistic agents particularly in immunocompromised patients. It is one of the top three pathogens causing many healthcare associated infections (HAIs). Resistance to several commonly used antimicrobial agents is a remarkable characteristic of most species which may carry various genes contributing to virulence. Objectives: to determine the prevalence of enterococci species in different intensive care units (ICUs) causing health care-associated infections (HAIs), intestinal carriage and environmental contamination. Also, to study the antimicrobial susceptibility pattern of the isolates with special reference to vancomycin resistance. In addition to phenotypic and genotypic detection of gelatinase, cytolysin and biofilm formation among isolates. Patients and Methods: This study was carried out in the infection control laboratory at Assiut University Hospitals over a period of one year. Clinical samples were collected from 285 patients with various (HAIs) acquired after admission to different ICUs. Rectal swabs were taken from 14 cases for detection of enterococci carriage. In addition, 1377 environmental samples were collected from the surroundings of the patients. Identification was done by conventional bacteriological methods and confirmed by analytical profile index (API). Antimicrobial sensitivity testing was performed by Kirby Bauer disc diffusion method and detection of vancomycin resistance was done by agar screen method. For the isolates, phenotypic detection of cytolysin, gelatinase production and detection of biofilm by tube method, Congo red method and microtiter plate. We performed polymerase chain reaction (PCR) for detection of some virulence genes (gelE, cylA, vanA, vanB and esp). Results: Enterococci caused 10.5% of the HAIs. Respiratory tract infection was the predominant type (86.7%). The commonest species were E.gallinarum (36.7%), E.casseliflavus (30%), E.faecalis (30%), and E.durans (3.4 %). Vancomycin resistance was detected in a total of 40% (12/30) of those isolates. The risk factors associated with acquiring vancomycin resistant enterococci (VRE) were immune suppression (P= 0.031) and artificial feeding (P= 0.008). For the rectal swabs, enterococci species were detected in 71.4% of samples with the predominance of E. casseliflavus (50%). Most of the isolates were vancomycin resistant (70%). Out of a total 1377 environmental samples, 577 (42%) samples were contaminated with different microorganisms. Enterococci were detected in 1.7% (10/577) of total contaminated samples, 50% of which were vancomycin resistant. All isolates were resistant to penicillin, ampicillin, oxacillin, ciprofloxacin, amikacin, erythromycin, clindamycin and trimethoprim-sulfamethaxazole. For the remaining antibiotics, variable percentages of resistance were reported. Cytolysin and gelatinase were detected phenotypically in 16% and 48 % of the isolates respectively. The microtiter plate method showed the highest percentages of detection of biofilm among all isolated species (100%). The studied virulence genes gelE, esp, vanA and vanB were detected in 62%, 12%, 2% and 12% respectively, while cylA gene was not detected in any isolates. Conclusions: A significant percentage of enterococci was isolated from patients and environments in the ICUs. Many virulence factors were detected phenotypically and genotypically among isolates. The high percentage of resistance, coupled with the risk of cross transmission to other patients make enterococci infections a significant infection control issue in hospitals.

Keywords: antimicrobial resistance, enterococci, ICUs, virulence factors

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Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

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Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

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Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri

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The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.

Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq

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Authors: Jaco Oosthuizen, Nerina C. Van Der Merwe

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The National Health Laboratory Services in Bloemfontien has been the diagnostic testing facility for 1830 patients for familial breast cancer since 1997. From the cohort, 540 were comprehensively screened using High-Resolution Melting Analysis or Next Generation Sequencing for the presence of point mutations and/or indels. Approximately 90% of these patients stil remain undiagnosed as they are BRCA1/2 negative. Multiplex ligation-dependent probe amplification was initially added to screen for copy number variation detection, but with the introduction of next generation sequencing in 2017, was substituted and is currently used as a confirmation assay. The aim was to investigate the viability of utilizing internationally designed copy number variation detection assays based on mostly European/Caucasian genomic data for use within a South African context. The multiplex ligation-dependent probe amplification technique is based on the hybridization and subsequent ligation of multiple probes to a targeted exon. The ligated probes are amplified using conventional polymerase chain reaction, followed by fragment analysis by means of capillary electrophoresis. The experimental design of the assay was performed according to the guidelines of MRC-Holland. For BRCA1 (P002-D1) and BRCA2 (P045-B3), both multiplex assays were validated, and results were confirmed using a secondary probe set for each gene. The next generation sequencing technique is based on target amplification via multiplex polymerase chain reaction, where after the amplicons are sequenced parallel on a semiconductor chip. Amplified read counts are visualized as relative copy numbers to determine the median of the absolute values of all pairwise differences. Various experimental parameters such as DNA quality, quantity, and signal intensity or read depth were verified using positive and negative patients previously tested internationally. DNA quality and quantity proved to be the critical factors during the verification of both assays. The quantity influenced the relative copy number frequency directly whereas the quality of the DNA and its salt concentration influenced denaturation consistency in both assays. Multiplex ligation-dependent probe amplification produced false positives due to ligation failure when ligation was inhibited due to a variant present within the ligation site. Next generation sequencing produced false positives due to read dropout when primer sequences did not meet optimal multiplex binding kinetics due to population variants in the primer binding site. The analytical sensitivity and specificity for the South African population have been proven. Verification resulted in repeatable reactions with regards to the detection of relative copy number differences. Both multiplex ligation-dependent probe amplification and next generation sequencing multiplex panels need to be optimized to accommodate South African polymorphisms present within the genetically diverse ethnic groups to reduce the false copy number variation positive rate and increase performance efficiency.

Keywords: familial breast cancer, multiplex ligation-dependent probe amplification, next generation sequencing, South Africa

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Authors: Aurora Sheehy, Caitlin Prince

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Within current orthodox medical protocols, trauma and mental health issues are deemed to reside within the realm of cognitive or psychological therapists and are marginalised in these areas, in part due to limited drugs option available, mostly manipulating neurotransmitters or sedating patients to reduce symptoms. By contrast, this research presents examples from the clinical practice of how trauma can be assessed and treated physiologically. Adverse Childhood Experiences (ACEs) are a tally of different types of abuse and neglect. It has been used as a measurable and reliable predictor of the likelihood of the development of autoimmune disease. It is a direct way to demonstrate reliably the health impact of traumatic life experiences. A second assessment tool is Allostatic Load, which refers to the cumulative effects that chronic stress has on mental and physical health. It records the decline of an individual’s physiological capacity to cope with their experience. It uses a specific grouping of serum testing and physical measures. It includes an assessment of neuroendocrine, cardiovascular, immune and metabolic systems. Allostatic load demonstrates the health impact that trauma has throughout the body. It forms part of an initial intake assessment in clinical practice and could also be used in research to evaluate treatment. Examining medicinal plants for their physiological, neurological and somatic effects through the lens of Polyvagal theory offers new opportunities for trauma treatments. In situations where Polyvagal theory recommends activities and exercises to enable parasympathetic activation, many herbs that affect Effector Memory T (TEM) cells also enact these responses. Traditional or Indigenous European herbs show the potential to support the polyvagal tone, through multiple mechanisms. As the ventral vagal nerve reaches almost every major organ, plants that have actions on these tissues can be understood via their polyvagal actions, such as monoterpenes as agents to improve respiratory vagal tone, cyanogenic glycosides to reset polyvagal tone, volatile oils rich in phenyl methyl esters improve both sympathetic and parasympathetic tone, bitters activate gut function and can strongly promote parasympathetic regulation. Auricular Acupuncture uses a system of somatotopic mapping of the auricular surface overlaid with an image of an inverted foetus with each body organ and system featured. Given that the concha of the auricle is the only place on the body where the Vagus Nerve neurons reach the surface of the skin, several investigators have evaluated non-invasive, transcutaneous electrical nerve stimulation (TENS) at auricular points. Drawn from an interdisciplinary evidence base and developed through clinical practice, these assessment and treatment tools are examples of practitioners in the field innovating out of necessity for the best outcomes for patients. This paper draws on case studies to direct future research.

Keywords: polyvagal, auricular acupuncture, trauma, herbs

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Authors: Netalie Shloim, Brian Brown, Siobhan Hugh-Jones, Jane Plastow, Diana Setiyawati, Anna Madill

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In June 2021, the World Health Organization launched its guidance and technical packages on community mental health services, stressing a human rights-based approach to care. This initiative stems from an increasing acknowledgment of the role mental health plays in achieving the Sustainable Development Goals. Nevertheless, mental health remains a relatively neglected research area and the estimates for untreated mental disorders in low-and-middle-income countries (LMICs) are as high as 78% for adults. Moreover, the development sector and research programs too often side-line mental health as a privilege in the face of often immediate threats to life and livelihood. As a way of addressing this problem, this study aimed to examine past or ongoing GCRF projects to see if there were opportunities where mental health impact could have been achieved without compromising a study's main aim and without overburdening a project. Projects funded by the UKRI Global Challenges Research Fund (GCRF) were analyzed. This program was initiated in 2015 to support cutting-edge research that addresses the challenges faced by developing countries. By the end of May 2020, a total of 15,279 projects were funded of which only 3% had an explicit mental health focus. A sample of 36 non-mental-health-focused projects was then sampled for diversity across research council, challenge portfolio and world region. Each of these 36 projects was coded by two coders for opportunities to embed mental health impact. To facilitate coding, the literature was inspected for dimensions relevant to LMIC settings. Three main psychological and three main social dimensions were identified: promote a positive sense of self; promote positive emotions, safe expression and regulation of challenging emotions, coping strategies, and help-seeking; facilitate skills development; and facilitate community-building; preserve sociocultural identity; support community mobilization. Coding agreement was strong on missed opportunities for mental health impact on the three social dimensions: support community mobilization (92%), facilitate community building (83%), preserve socio-cultural identity (70%). Coding agreement was reasonably strong on missed opportunities for mental health impact on the three psychological dimensions: promote positive emotions (67%), facilitate skills development (61%), positive sense of self (58%). In order of frequency, the agreed perceived opportunities from the highest to lowest are: support community mobilization, facilitate community building, facilitate skills development, promote a positive sense of self, promote positive emotions, preserve sociocultural identity. All projects were considered to have an opportunity to support community mobilization and to facilitate skills development by at least one coder. Findings provided support that there were opportunities to embed mental health impact in research across the range of development sectors and identifies what kind of missed opportunities are most frequent. Hence, mainstreaming mental health has huge potential to tackle the lack of priority and funding it has attracted traditionally. The next steps are to understand the barriers to mainstreaming mental health and to work together to overcome them.

Keywords: GCRF, mental health, psychosocial wellbeing, LMIC

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Authors: Shpetim Rezniqi

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Financial Policies in the process of global crisis the current crisis has swept the world with special emphasis, most developed countries, those countries which have most gross -product world and you have a high level of living.Even those who are not experts can describe the consequences of the crisis to see the reality that is seen, but how far will it go this crisis is impossible to predict. Even the biggest experts have conjecture and large divergence, but agree on one thing: - The devastating effects of this crisis will be more severe than ever before and can not be predicted.Long time, the world was dominated economic theory of free market laws. With the belief that the market is the regulator of all economic problems. The market, as river water will flow to find the best and will find the necessary solution best. Therefore much less state market barriers, less state intervention and market itself is an economic self-regulation. Free market economy became the model of global economic development and progress, it transcends national barriers and became the law of the development of the entire world economy. Globalization and global market freedom were principles of development and international cooperation. All international organizations like the World Bank, states powerful economic, development and cooperation principles laid free market economy and the elimination of state intervention. The less state intervention much more freedom of action was this market- leading international principle. We live in an era of financial tragic. Financial markets and banking in particular economies are in a state of thy good, US stock markets fell about 40%, in other words, this time, was one of the darkest moments 5 since 1920. Prior to her rank can only "collapse" of the stock of Wall Street in 1929, technological collapse of 2000, the crisis of 1973 after the Yom Kippur war, while the price of oil quadrupled and famous collapse of 1937 / '38, when Europe was beginning World war II In 2000, even though it seems like the end of the world was the corner, the world economy survived almost intact. Of course, that was small recessions in the United States, Europe, or Japan. Much more difficult the situation was at crisis 30s, or 70s, however, succeeded the world. Regarding the recent financial crisis, it has all the signs to be much sharper and with more consequences. The decline in stock prices is more a byproduct of what is really happening. Financial markets began dance of death with the credit crisis, which came as a result of the large increase in real estate prices and household debt. It is these last two phenomena can be matched very well with the gains of the '20s, a period during which people spent fists as if there was no tomorrow. All is not away from the mouth of the word recession, that fact no longer a sudden and abrupt. But as much as the financial markets melt, the greater is the risk of a problematic economy for years to come. Thus, for example, the banking crisis in Japan proved to be much more severe than initially expected, partly because the assets which were based more loans had, especially the land that falling in value. The price of land in Japan is about 15 years that continues to fall. (ADRI Nurellari-Published in the newspaper "Classifieds"). At this moment, it is still difficult to çmosh to what extent the crisis has affected the economy and what would be the consequences of the crisis. What we know is that many banks will need more time to reduce the award of credit, but banks have this primary function, this means huge loss.

Keywords: globalisation, finance, crisis, recomandation, bank, credits

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Authors: Michael Picard

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This proposal offers to shed light on the changing legal geography of the global waste economy. Global waste recycling has become a multi-billion-dollar industry. NASDAQ predicts the emergence of a worldwide 1,296G$ waste management market between 2017 and 2022. Underlining this evolution, a new generation of preferential waste-trade agreements has emerged in the Pacific. In the last decade, Japan has concluded a series of bilateral treaties with Asian countries, and most recently with China. An agreement between Tokyo and Beijing was formalized on 7 May 2008, which forged an economic partnership on waste transfer and mining. The agreement set up International Recycling Zones, where certified recycling plants in China process industrial waste imported from Japan. Under the joint venture, Chinese companies salvage the embedded value from Japanese industrial discards, reprocess them and send them back to Japanese manufacturers, such as Mitsubishi and Panasonic. This circular economy is designed to convert surplus garbage into surplus value. Ever since the opening of Sino-Japanese eco-parks, millions of tons of plastic and e-waste have been exported from Japan to China every year. Yet, quite unexpectedly, China has recently closed its waste market to imports, jeopardizing Japan’s billion-dollar exports to China. China notified the WTO that, by the end of 2017, it would no longer accept imports of plastics and certain metals. Given China’s share of Japanese waste exports, a complete closure of China’s market would require Japan to find new uses for its recyclable industrial trash generated domestically every year. It remains to be seen how China will effectively implement its ban on waste imports, considering the economic interests at stake. At this stage, what remains to be clarified is whether China's ban on waste imports will negatively affect the recycling trade between Japan and China. What is clear, though, is the rapid transformation in the legal geography of waste mining in East-Asia. For decades, East-Asian waste trade had been tied up in an ‘ecologically unequal exchange’ between the Japanese core and the Chinese periphery. This global unequal waste distribution could be measured by the Environmental Stringency Index, which revealed that waste regulation was 39% weaker in the Global South than in Japan. This explains why Japan could legally export its hazardous plastic and electronic discards to China. The asymmetric flow of hazardous waste between Japan and China carried the colonial heritage of international law. The legal geography of waste distribution was closely associated to the imperial construction of an ecological trade imbalance between the Japanese source and the Chinese sink. Thus, China’s recent decision to ban hazardous waste imports is a sign of a broader ecological shift. As a global economic superpower, China announced to the world it would no longer be the planet’s junkyard. The policy change will have profound consequences on the global circulation of waste, re-routing global waste towards countries south of China, such as Vietnam and Malaysia. By the time the Berlin Conference takes place in May 2018, the presentation will be able to assess more accurately the effect of the Chinese ban on the transboundary movement of waste in Asia.

Keywords: Asia, ecological unequal exchange, global waste trade, legal geography

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Authors: Hülya Torun, Ondřej Novák, Jaromír Mikulík, Miroslav Strnad, Faik A. Ayaz

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World climate is changing. Millions of people in the world still face chronic undernourishment for conducting a healthy life and the world’s population is growing steadily. To meet this growing demand, agriculture and food systems must adapt to the adverse effects of climate change and become more resilient, productive and sustainable. From this perspective, to determine tolerant cultivars for undesirable environmental conditions will be necessary food production for sustainable development. Among abiotic stresses, soil salinity is one of the most detrimental global fact restricting plant sources. Development of salt-tolerant lines is required in order to increase the crop productivity and quality in salt-treated lands. Therefore, the objective of this study was to investigate the morphological and physiological responses of barley cultivars accessions to salinity stress by NaCl. For this purpose, it was aimed to determine the crosstalk between some endogenous phytohormones and exogenous salicylic acid (SA) in two different vegetative parts (leaves and roots) of barley (Hordeum vulgare L.; Poaceae; 2n=14; Ince-04) which is detected salt-tolerant. The effects of SA on growth parameters, leaf relative water content (RWC), endogenous phytohormones; including indole-3-acetic acid (IAA), cytokinins (CKs), abscisic acid (ABA), jasmonic acid (JA) and ethylene were investigated in barley cultivars under salinity stress. SA was applied to 17-day-old seedlings of barley in two different ways including before (pre-treated for 24 h) and simultaneously with NaCl stress treatment. NaCl (0, 150, 300 mM) exposure in the hydrophonic system was associated with a rapid decrease in growth parameters and RWC, which is an indicator of plant water status, resulted in a strong up-regulation of ABA as a stress indicator. Roots were more dramatically affected than leaves. Water conservation in 150 mM NaCl treated-barley plants did not change, but decreased in 300 mM NaCl treated plants. Pre- and simultaneously treatment of SA did not significantly alter growth parameters and RWC. ABA, JA and ethylene are known to be related with stress. In the present work, ethylene also increased, similarly to ABA, but not with the same intensity. While ABA and ethylene increased by the increment of salt concentrations, JA levels rapidly decreased especially in roots. Both pre- and simultaneously SA applications alleviated salt-induced decreases in 300 mM NaCl resulted in the increment of ABA levels. CKs and IAA are related to cell growth and development. At high salinity (300 mM NaCl), CKs (cZ+cZR) contents increased in both vegetative organs while IAA levels stayed at the same level with control groups. However, IAA increased and cZ+cZR rapidly decreased in leaves of barley plants with SA treatments before salt applications (in pre- SA treated groups). Simultaneously application of SA decreased CKs levels in both leaves and roots of the cultivar. Due to increasing concentrations of NaCl in association with decreasing ABA, JA and ethylene content and increments in CKs and IAA were recorded with SA treatments. As results of the study, in view of all the phytohormones that we tested, exogenous SA induced greater tolerance to salinity particularly when applied before salinity stress.

Keywords: Barley, Hordeum vulgare, phytohormones, salicylic acid, salinity

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Authors: Swati Gautam, Salman Akhtar, S. P. Jaiswar, Amita Jain

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Background: FGTB is now a major global health problem mostly in developing countries including India. In humans, Mycobacterium Tuberculosis (M.tb) is a causating agent of infection. High index of suspicion is required for early diagnosis due to asymptomatic presentation of FGTB disease. In macrophages Toll Like Receptor-2 (TLR-2) is one which mediated host’s immune response to M.tb. The expression of TLR-2 on macrophages is important to determine the fate of innate immune responses to M.tb. TLR-2 have two work. First its high expression on macrophages worsen the outer of infection and another side, it maintains M.tb to its dormant stage avoids activation of M.tb from latent phase. Single Nucleotide Polymorphism (SNP) of TLR-2 gene plays an important role in susceptibility to TB among different populations and subsequently, in the development of infertility. Methodology: This Case-Control study was done in the Department of Obs and Gynae and Department of Microbiology at King George’s Medical University, U.P, Lucknow, India. Total 300 subjects (150 Cases and 150 Controls) were enrolled in the study. All subjects were enrolled only after fulfilling the given inclusion and exclusion criteria. Inclusion criteria: Age 20-35 years, menstrual-irregularities, positive on Acid-Fast Bacilli (AFB), TB-PCR, (LJ/MGIT) culture in Endometrial Aspiration (EA). Exclusion criteria: Koch’s active, on ATT, PCOS, and Endometriosis fibroid women, positive on Gonococal and Chlamydia. Blood samples were collected in EDTA tubes from cases and healthy control women (HCW) and genomic DNA extraction was carried out by salting-out method. Genotyping of TLR2 genetic variants (Arg753Gln and Arg677Trp) were performed by using single amplification refractory mutation system (ARMS) PCR technique. PCR products were analyzed by electrophoresis on 1.2% agarose gel and visualized by gel-doc. Statistical analysis of the data was performed using the SPSS 16.3 software and computing odds ratio (OR) with 95% CI. Linkage Disequiliribium (LD) analysis was done by SNP stats online software. Results: In TLR-2 (Arg753Gln) polymorphism significant risk of FGTB observed with GG homozygous mutant genotype (OR=13, CI=0.71-237.7, p=0.05), AG heterozygous mutant genotype (OR=13.7, CI=0.76-248.06, p=0.03) however, G allele (OR=1.09, CI=0.78-1.52, p=0.67) individually was not associated with FGTB. In TLR-2 (Arg677Trp) polymorphism a significant risk of FGTB observed with TT homozygous mutant genotype (OR= 0.020, CI=0.001-0.341, p < 0.001), CT heterozygous mutant genotype (OR=0.53, CI=0.33-0.86, p=0.014) and T allele (OR=0.463, CI=0.32-0.66, p < 0.001). TT mutant genotype was only found in FGTB cases and frequency of CT heterozygous more in control group as compared to FGTB group. So, CT genotype worked as protective mutation for FGTB susceptibility group. In haplotype analysis of TLR-2 genetic variants, four possible combinations, i.e. (G-T, A-C, G-C, and A-T) were obtained. The frequency of haplotype A-C was significantly higher in FGTB cases (0.32). Control group did not show A-C haplotype and only found in FGTB cases. Conclusion: In conclusion, study showed a significant association with both genetic variants of TLR-2 of FGTB disease. Moreover, the presence of specific associated genotype/alleles suggest the possibility of disease severity and clinical approach aimed to prevent extensive damage by disease and also helpful for early detection of disease.

Keywords: ARMS, EDTA, FGTB, TLR

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Authors: Qudsiya Baig

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Empathy yields an unparalleled therapeutic value within patient physician interactions. Medical research is inundated with evidence to support that a physician’s ability to empathise with patients leads to a greater willingness to report symptoms, an improvement in diagnostic accuracy and safety, and a better adherence and satisfaction with treatment plans. Furthermore, the Institute of Medicine states that empathy leads to a more patient-centred care, which is one of the six main goals of a 21st century health system. However, there is a paradox between the theoretical significance of empathy and its presence, or lack thereof, in clinical practice. Recent studies have reported that empathy declines amongst students and physicians over time. The three most impactful contributors to this decline are: (1) disagreements over the definitions of empathy making it difficult to implement it into practice (2) poor consideration or regulation of empathy leading to burnout and thus, abandonment altogether, and (3) the lack of diversity in the curriculum and the influence of medical culture, which prioritises science over patient experience, limiting some physicians from using ‘too much’ empathy in the fear of losing clinical objectivity. These issues were investigated by conducting a fully inductive thematic narrative analysis of patient narratives in the UK to evaluate the behaviours and attitudes that patients associate with empathy. The principal enquiries underpinning this study included uncovering the factors that affected experience of empathy within provider-patient interactions and to analyse their effects on patient care. This research contributes uniquely to this discourse by examining the phenomenon of empathy directly from patients’ experiences, which were systematically extracted from a repository of online patient narratives of care titled ‘CareOpinion UK’. Narrative analysis was specifically chosen as the methodology to examine narratives from a phenomenological lens to focus on the particularity and context of each story. By enquiring beyond the superficial who-whatwhere, the study of narratives prescribed meaning to illness by highlighting the everyday reality of patients who face the exigent life circumstances created by suffering, disability, and the threat of life. The following six themes were found to be the most impactful in influencing the experience of empathy: dismissive behaviours, judgmental attitudes, undermining patients’ pain or concerns, holistic care and failures and successes of communication or language. For each theme there were overarching themes relating to either a failure to understand the patient’s perspective or a success in taking a person-centred approach. An in-depth analysis revealed that a lack of empathy was greatly associated with an emotive-cognitive imbalance, which disengaged physicians with their patients’ emotions. This study hereby concludes that competent providers require a combination of knowledge, skills, and more importantly empathic attitudes to help create a context for effective care. The crucial elements of that context involve (a) identifying empathy clues within interactions to engage with patients’ situations, (b) attributing a perspective to the patient through perspective-taking and (c) adapting behaviour and communication according to patient’s individual needs. Empathy underpins that context, as does an appreciation of narrative, and the two are interrelated.

Keywords: empathy, narratives, person-centred, perspective, perspective-taking

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Authors: A. G. Back

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Urban planning is a relevant tool to address, in a systemic way, several sectoral policies capable of linking the urban agenda with the reduction of socio-environmental risks. The Sao Paulo’s master plan (2014) presents innovations capable of promoting the transition to sustainability in the urban space, with a view to its regulatory instruments related to i) promotion of density in the axes of mass transport involving the mixture of commercial, residential, services, and leisure uses (principles related to the compact city); ii) vulnerabilities reduction based on housing policies including regular sources of funds for social housing and land reservation in urbanized areas; iii) reserve of green areas in the city to create parks and environmental regulations for new buildings focused on reducing the effects of heat island and improving urban drainage. However, its long-term implementation involves distributive conflicts and can undergo changes in different political, economic, and social contexts over time. Thus, the main objective of this paper is to identify and analyze the dynamics of conflicts of interest between social groups in the implementation of Sao Paulo’s urban development policy, particularly in relation to recent attempts at a (re) interpretation of the Master Plan guidelines, in view of the proposals for revision of the urban zoning law. In this sense, we seek to identify the demands, narratives of urban actors, including the real estate market, middle-class neighborhood associations ('not in my backyard' movements), and social housing rights movements. And we seek to analyze the power resources that these actors mobilize to influence the decision-making process, involving five categories: social capital, political access; discursive resource; media, juridical resource. The major findings of this research suggest that the interests and demands of the real estate market do not always prevail in urban regulation. After all, other actors also press for the definition of urban law with interests opposite to those of the real estate market. This is the case of associations of middle-class neighborhoods, which work to protect the characteristics of the locality, acting, in general, to prevent constructive and population densification in neighborhoods well located near the center, in São Paulo. One of the main demands of these “not in my backyard” movements is the delimitation of exclusively residential areas in the central region of the city, which is not only contrary to the interests of the real state market but also contrary to the principles of the compact city. On the other hand, social housing rights movements have also made progress in delimiting special areas of social interest in well-located and valued areas in the city dedicated to building social housing, also contrary to the interests of the real estate market. An urban development that follows the principles of the compact city must take into account the insertion of low-income populations in well-located regions; otherwise, such a development model may continue to push the less favored to the peripheries towards the preservation areas and/or risk areas.

Keywords: interest groups, Sao Paulo, sustainable urban development, urban policies implementation

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Authors: Prajna Paramita Naik, Sujit Kumar Bhutia

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Background: Regardless of the development multimodal treatment strategies, oral squamous cell carcinoma (OSCC) is often associated with a high rate of recurrence, metastasis and chemo- and radio- resistance. The present study inspected the relevance of CD44, ABCB1 and ADAM17 expression as a putative stem cell compartment in oral squamous cell carcinoma (OSCC) and deciphered the role of autophagy in regulating the expression of aforementioned proteins, stemness and chemoresistance. Methods: A retrospective analysis of CD44, ABCB1 and ADAM17 expression with respect to the various clinicopathological factors of sixty OSCC patients were determined via immunohistochemistry. The correlation among CD44, ABCB1 and ADAM17 expression was established. Sphere formation assay, flow cytometry and fluorescence microscopy were conducted to elucidate the stemness and chemoresistance nature of established cisplatin-resistant oral cancer cells (FaDu). The pattern of expression of CD44, ABCB1 and ADAM17 in parental (FaDu-P) and resistant FaDu cells (FaDu-CDDP-R) were investigated through fluorescence microscopy. Western blot analysis of autophagy marker proteins was performed to compare the status of autophagy in parental and resistant FaDu cell. To investigate the role of autophagy in chemoresistance and stemness, sphere formation assay, immunofluorescence and Western blot analysis was performed post transfection with siATG14 and the level of expression of autophagic proteins, mitochondrial protein and stemness-associated proteins were analyzed. The statistical analysis was performed by GraphPad Prism 4.0 software. p-value was defined as follows: not significant (n.s.): p > 0.05;*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001; ****: p ≤ 0.0001 were considered statistically significant. Results: In OSCC, high CD44, ABCB1 and ADAM17 expression were significantly correlated with higher tumor grades and poor differentiation. However, the expression of these proteins was not related to the age and sex of OSCC patients. Moreover, the expression of CD44, ABCB1 and ADAM17 were positively correlated with each other. In vitro and OSCC tissue double labeling experiment data showed that CD44+ cells were highly associated with ABCB1 and ADAM17 expression. Further, FaDu-CDDP-R cells showed higher sphere forming capacity along with increased fraction of the CD44+ population and β-catenin expression FaDu-CDDP-R cells also showed accelerated expression of CD44, ABCB1 and ADAM17. A comparatively higher autophagic flux was observed in FaDu-CDDP-R against FaDu-P cells. The expression of mitochondrial proteins was noticeably reduced in resistant cells as compared to parental cells indicating the occurrence of autophagy-mediated mitochondrial degradation in oral cancer. Moreover, inhibition of autophagy was coupled with the decreased formation of orospheres suggesting autophagy-mediated stemness in oral cancer. Blockade of autophagy was also found to induce the restoration of mitochondrial proteins in FaDu-CDDP-R cells indicating the involvement of mitophagy in chemoresistance. Furthermore, a reduced expression of CD44, ABCB1 and ADAM17 was also observed in ATG14 deficient cells FaDu-P and FaDu-CDDP-R cells. Conclusion: The CD44+ ⁄ABCB1+ ⁄ADAM17+ expression in OSCC might be associated with chemoresistance and a putative CSC compartment. Further, the present study highlights the contribution of mitophagy in chemoresistance and confirms the potential involvement of autophagic regulation in acquisition of stem-like characteristics in OSCC.

Keywords: ABCB1, ADAM17, autophagy, CD44, chemoresistance, mitophagy, OSCC, stemness

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Authors: S. N. T. I. Sampath, J. M. S. Jayasinghe, A. P. Attanayake, V. Karunaratne

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Diabetes mellitus is one of the major public health posers throughout the world today that incidence and associated with increasing mortality. Insufficient regulation of the blood glucose level might be serious effects for health and its necessity to identify new therapeutics that have ability to reduce hyperglycaemic condition in the human body. Even though synthetic antidiabetic drugs are more effective to control diabetes mellitus, there are considerable side effects have been reported. Thus, there is an increasing demand for searching new natural products having high antidiabetic activity with lesser side effects. The purposes of the present study were to evaluate different physical parameters and in-vitro and in-vivo antidiabetic potential of the selected combined medicinal plant extracts mixture composed of leaves of Murraya koenigii, cloves of Allium sativum, fruits of Garcinia queasita and seeds of Piper nigrum. The selected plants parts were mixed and ground together and extracted sequentially into the hexane, ethyl acetate and methanol. Solvents were evaporated and they were further dried by freeze-drying to obtain a fine powder of each extract. Various physical parameters such as moisture, total ash, acid insoluble ash and water soluble ash were evaluated using standard test procedures. In-vitro antidiabetic activity of combined plant extracts mixture was screened using enzyme assays such as α-amylase inhibition assay and α-glucosidase inhibition assay. The acute anti-hyperglycaemic activity was performed using oral glucose tolerance test for the streptozotocin induced diabetic Wistar rats to find out in-vivo antidiabetic activity of combined plant extracts mixture and it was assessed through total oral glucose tolerance curve (TAUC) values. The percentage of moisture content, total ash content, acid insoluble ash content and water soluble ash content were ranged of 7.6-17.8, 8.1-11.78, 0.019-0.134 and 6.2-9.2 respectively for the plant extracts and those values were less than standard values except the methanol extract. The hexane and ethyl acetate extracts exhibited highest α-amylase (IC50 = 25.7 ±0.6; 27.1 ±1.2 ppm) and α-glucosidase (IC50 = 22.4 ±0.1; 33.7 ±0.2 ppm) inhibitory activities than methanol extract (IC50 = 360.2 ±0.6; 179.6 ±0.9 ppm) when compared with the acarbose positive control (IC50 = 5.7 ±0.4; 17.1 ±0.6 ppm). The TAUC values for hexane, ethyl acetate, and methanol extracts and glibenclamide (positive control) treated rats were 8.01 ±0.66; 8.05 ±1.07; 8.40±0.50; 5.87 ±0.93 mmol/L.h respectively, whereas in diabetic control rats the TAUC value was 13.22 ±1.07 mmol/L.h. Administration of plant extracts treated rats significantly suppressed (p<0.05) the rise in plasma blood glucose levels compared to control rats but less significant than glibenclamide. The obtained results from in-vivo and in-vitro antidiabetic study showed that the hexane and ethyl acetate extracts of selected combined plant mixture might be considered as a potential source to isolate natural antidiabetic agents and physical parameters of hexane and ethyl acetate extracts will helpful to develop antidiabetic drug with further standardize properties.

Keywords: diabetes mellitus, in-vitro antidiabetic assays, medicinal plants, standardization

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Authors: Ting Wu, Feiting Hu, Xinyue Zhang, Shuxin Tang, Xiaoyun Xu

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As a by-product of yuba, processing wastewater of Yuba (PWY) contains many bioactive components such as soybean isoflavones, soybean polysaccharides and soybean oligosaccharides, which is a good source of prebiotics and has a potential of high value utilization. The use of Lactobacillus plantarum to ferment PWY can be considered as a potential biogenic element, which can regulate the balance of intestinal microbiota. In this study, firstly, Lactobacillus plantarum was used to ferment PWY to improve its content of active components and antioxidant activity. Then, the health effect of fermented processing wastewater of yuba (FPWY) was measured in vitro. Finally, microencapsulation technology was used applied to improve the sustained release of FPWY and reduce the loss of active components in the digestion process, as well as to improving the activity of FPWY. The main results are as follows: (1) FPWY presented a good antioxidant capacity with DPPH free radical scavenging ability (0.83 ± 0.01 mmol Trolox/L), ABTS free radical scavenging ability (7.47 ± 0.35 mmol Trolox/L) and iron ion reducing ability (1.11 ± 0.07 mmol Trolox/L). Compared with non-fermented processing wastewater of yuba (NFPWY), there was no significant difference in the content of total soybean isoflavones, but the content of glucoside soybean isoflavones decreased, and aglyconic soybean isoflavones increased significantly. After fermentation, PWY can effectively reduce the soluble monosaccharides, disaccharides and oligosaccharides, such as glucose, fructose, galactose, trehalose, stachyose, maltose, raffinose and sucrose. (2) FPWY can significantly enhance the growth of beneficial bacteria such as Bifidobacterium, Ruminococcus and Akkermansia, significantly inhibit the growth of harmful bacteria E.coli, regulate the structure of intestinal microbiota, and significantly increase the content of short-chain fatty acids such as acetic acid, propionic acid, butyric acid, isovaleric acid. Higher amount of lactic acid in the gut can be further broken down into short chain fatty acids. (3) In order to improve the stability of soybean isoflavones in FPWY during digestion, sodium alginate and chitosan were used as wall materials for embedding. The FPWY freeze-dried powder was embedded by the method of acute-coagulation bath. The results show that when the core wall ratio is 3:1, the concentration of chitosan is 1.5%, the concentration of sodium alginate is 2.0%, and the concentration of calcium is 3%, the embossing rate is 53.20%. In the simulated in vitro digestion stage, the release rate of microcapsules reached 59.36% at the end of gastric digestion and 82.90% at the end of intestinal digestion. Therefore, the core materials with good sustained-release performance of microcapsules were almost all released. The structural analysis results of FPWY microcapsules show that the microcapsules have good mechanical properties. Its hardness, springness, cohesiveness, gumminess, chewiness and resilience were 117.75± 0.21 g, 0.76±0.02, 0.54±0.01, 63.28±0.71 g·sec, 48.03±1.37 g·sec, 0.31±0.01, respectively. Compared with the unembedded FPWY, the infrared spectrum results showed that the microcapsules had embedded effect on the FPWY freeze-dried powder.

Keywords: processing wastewater of yuba, lactobacillus plantarum, intestinal microbiota, microcapsule

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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Authors: Jacqueline Eleonora Ek, Michael Spiteri, Chris Giordimaina, Pierre Agius

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Background: Malta is known for being a key player as a frontline country with regard to irregular immigration from Africa to Europe. Every year the island experiences an influx of migrants as boat movement across the Mediterranean continues to be a humanitarian challenge. Irregular immigration and applying for asylum is both a lengthy and mentally demanding process. Those doing so are often faced with multiple challenges, which can adversely affect their mental health. Between January and August 2020, Malta disembarked 2 162 people rescued at sea, 463 of them between July & August. Given the small size of the Maltese islands, this regulation places a disproportionately large burden on the country, creating a backlog in the processing of asylum applications resulting in increased time periods of detention. These delays reverberate throughout multiple management pathways resulting in prolonged periods of detention and challenging access to health services. Objectives: To better understand the spatial dimensions of this humanitarian crisis, this study aims to assess disparities in the acute medical management of migrants presenting to the emergency department (ED) with acute mental health presentations as compared to that of local and non-local residents. Method: In this retrospective study, 17795 consecutive ED attendances were reviewed to look for acute mental health presentations. These were further evaluated to assess discrepancies in transportation routes to hospital, nature of presenting complaint, effects of language barriers, use of CT brain, treatment given at ED, availability of psychiatric reviews, and final admission/discharge plans. Results: Of the ED attendances, 92.3% were local residents, and 7.7% were non-locals. Of the non-locals, 13.8% were migrants, and 86.2% were other-non-locals. Acute mental health presentations were seen in 1% of local residents; this increased to 20.6% in migrants. 56.4% of migrants attended with deliberate self-harm; this was lower in local residents, 28.9%. Contrastingly, in local residents, the most common presenting complaint was suicidal thought/ low mood 37.3%, the incidence was similar in migrants at 33.3%. The main differences included 12.8% of migrants presenting with refused oral intake while only 0.6% of local residents presented with the same complaints. 7.7% of migrants presented with a reduced level of consciousness, no local residents presented with this same issue. Physicians documented a language barrier in 74.4% of migrants. 25.6% were noted to be completely uncommunicative. Further investigations included the use of a CT scan in 12% of local residents and in 35.9% of migrants. The most common treatment administered to migrants was supportive fluids 15.4%, the most common in local residents was benzodiazepines 15.1%. Voluntary psychiatric admissions were seen in 33.3% of migrants and 24.7% of locals. Involuntary admissions were seen in 23% of migrants and 13.3% of locals. Conclusion: Results showed multiple disparities in health management. A meeting was held between entities responsible for migrant health in Malta, including the emergency department, primary health care, migrant detention services, and Malta Red Cross. Currently, national quality-improvement initiatives are underway to form new pathways to improve patient-centered care. These include an interpreter unit, centralized handover sheets, and a dedicated migrant health service.

Keywords: emergency department, communication, health, migration

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Authors: Alena Nesterenko, Svetlana Petrikova

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Research evaluation is one of the most important elements of self-regulation and development of researchers as it is impartial and independent process of assessment. The method of expert evaluations as a scientific instrument solving complicated non-formalized problems is firstly a scientifically sound way to conduct the assessment which maximum effectiveness of work at every step and secondly the usage of quantitative methods for evaluation, assessment of expert opinion and collective processing of the results. These two features distinguish the method of expert evaluations from long-known expertise widespread in many areas of knowledge. Different typical problems require different types of expert evaluations methods. Several issues which arise with these methods are experts’ selection, management of assessment procedure, proceeding of the results and remuneration for the experts. To address these issues an on-line system was created with the primary purpose of development of a versatile application for many workgroups with matching approaches to scientific work management. Online documentation assessment and statistics system allows: - To realize within one platform independent activities of different workgroups (e.g. expert officers, managers). - To establish different workspaces for corresponding workgroups where custom users database can be created according to particular needs. - To form for each workgroup required output documents. - To configure information gathering for each workgroup (forms of assessment, tests, inventories). - To create and operate personal databases of remote users. - To set up automatic notification through e-mail. The next stage is development of quantitative and qualitative criteria to form a database of experts. The inventory was made so that the experts may not only submit their personal data, place of work and scientific degree but also keywords according to their expertise, academic interests, ORCID, Researcher ID, SPIN-code RSCI, Scopus AuthorID, knowledge of languages, primary scientific publications. For each project, competition assessments are processed in accordance to ordering party demands in forms of apprised inventories, commentaries (50-250 characters) and overall review (1500 characters) in which expert states the absence of conflict of interest. Evaluation is conducted as follows: as applications are added to database expert officer selects experts, generally, two persons per application. Experts are selected according to the keywords; this method proved to be good unlike the OECD classifier. The last stage: the choice of the experts is approved by the supervisor, the e-mails are sent to the experts with invitation to assess the project. An expert supervisor is controlling experts writing reports for all formalities to be in place (time-frame, propriety, correspondence). If the difference in assessment exceeds four points, the third evaluation is appointed. As the expert finishes work on his expert opinion, system shows contract marked ‘new’, managers commence with the contract and the expert gets e-mail that the contract is formed and ready to be signed. All formalities are concluded and the expert gets remuneration for his work. The specificity of interaction of the examination officer with other experts will be presented in the report.

Keywords: expertise, management of research evaluation, method of expert evaluations, research evaluation

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Authors: Rebekah Granger Ellis, Richard B. Speaker, Pat Austin

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This study used two psychological scales to examine the level of social and emotional intelligence and moral judgment of over 500 gifted and talented high school students in various academic and creative arts programs in a large metropolitan area in the southeastern United States. For decades, numerous models and programs purporting to encourage socio-affective characteristics of adolescent development have been explored in curriculum theory and design. Socio-affective merges social, emotional, and moral domains. It encompasses interpersonal relations and social behaviors; development and regulation of emotions; personal and gender identity construction; empathy development; moral development, thinking, and judgment. Examining development in these socio-affective domains can provide insight into why some gifted and talented adolescents are not successful in adulthood despite advanced IQ scores. Particularly whether nonintellectual characteristics of gifted and talented individuals, such as emotional, social and moral capabilities, are as advanced as their intellectual abilities and how these are related to each other. Unique characteristics distinguish gifted and talented individuals; these may appear as strengths, but there is the potential for problems to accompany them. Although many thrive in their school environments, some gifted students struggle rather than flourish. In the socio-affective domain, these adolescents face special intrapersonal, interpersonal, and environmental problems. Gifted individuals’ cognitive, psychological, and emotional development occurs asynchronously, in multidimensional layers at different rates and unevenly across ability levels. Therefore, it is important to examine the long-term effects of participation in various gifted and talented programs on the socio-affective development of gifted and talented adolescents. This quasi-experimental longitudinal study examined students in several gifted and talented education programs (creative arts school, urban charter schools, and suburban public schools) for (1) socio-affective development level and (2) whether a particular gifted and talented program encourages developmental growth. The following research questions guided the study: (1) How do academically and artistically talented gifted 10th and 11th grade students perform on psychometric scales of social and emotional intelligence and moral judgment? Do they differ from their age or grade normative sample? Are their gender differences among gifted students? (2) Does school environment impact 10th and 11th grade gifted and talented students’ socio-affective development? Do gifted adolescents who participate in a particular school gifted program differ in their developmental profiles of social and emotional intelligence and moral judgment? Students’ performances on psychometric instruments were compared over time and by type of program. Participants took pre-, mid-, and post-tests over the course of an academic school year with Defining Issues Test (DIT-2) assessing moral judgment and BarOn EQ-I: YV assessing social and emotional intelligence. Based on these assessments, quantitative differences in growth on psychological scales (individual and school) were examined. Change scores between schools were also compared. If a school showed change, artifacts (culture, curricula, instructional methodology) provided insight as to environmental qualities that produced this difference.

Keywords: gifted and talented education, moral development, socio-affective development, socio-affective education

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Authors: Harry Einhorn

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This paper aims to explore the integration of Buddhism into educational settings with the goal of fostering the holistic development of children. By incorporating Buddhist principles and practices, educators can create a nurturing environment that cultivates wisdom, compassion, and ethical values in children. The teachings of Buddhism provide valuable insights into mindfulness, compassion, and critical thinking, which can be adapted and applied to educational curricula to enhance children's intellectual, emotional, and moral growth. One of the fundamental aspects of Buddhist philosophy that is particularly relevant to education is the concept of mindfulness. By introducing mindfulness practices, such as meditation and breathing exercises, children can learn to cultivate present-moment awareness, develop emotional resilience, and enhance their ability to concentrate and focus. These skills are essential for effective learning and can contribute to reducing stress and promoting overall well-being in children. Mindfulness practices can also teach children how to manage their emotions and thoughts, promoting self-regulation and creating a positive classroom environment. In addition to mindfulness, Buddhism emphasizes the cultivation of compassion and empathy toward all living beings. Integrating teachings on kindness, empathy, and ethical behavior into the educational framework can help children develop a deep sense of interconnectedness and social responsibility. By engaging children in activities that promote empathy and encourage acts of kindness, such as community service projects and cooperative learning, educators can foster the development of compassionate individuals who are actively engaged in creating a more harmonious and compassionate society. Moreover, Buddhist teachings encourage critical thinking and inquiry, which are crucial skills for intellectual development. By introducing children to fundamental Buddhist concepts such as impermanence, interdependence, and the nature of suffering, educators can engage them in philosophical reflections and broaden their perspectives on life. These teachings promote open-mindedness, curiosity, and a deeper understanding of the interconnectedness of all things. Through the exploration of these concepts, children can develop critical thinking skills and gain insights into the complexities of the world, enabling them to navigate challenges with wisdom and discernment. While integrating Buddhism into education requires sensitivity, cultural awareness, and respect for diverse beliefs and backgrounds, it holds great potential for nurturing the holistic development of children. By incorporating mindfulness practices, fostering compassion and empathy, and promoting critical thinking, Buddhism can contribute to the creation of a more compassionate, inclusive, and harmonious educational environment. This integration can shape well-rounded individuals who are equipped with the necessary skills and qualities to navigate the complexities of the modern world with wisdom, compassion, and resilience. In conclusion, the integration of Buddhism into education offers a valuable framework for cultivating wisdom, compassion, and ethical values in children. By incorporating mindfulness, compassion, and critical thinking into educational practices, educators can create a supportive environment that promotes children's holistic development. By nurturing these qualities, Buddhism can help shape individuals who are not only academically proficient but also morally and ethically responsible, contributing to a more compassionate and harmonious society.

Keywords: Buddhism, education, children, mindfulness

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Authors: Omer Zvi Shaked, Amir Tirosh

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Type 1 Diabetes (T1DM) is an incurable chronic illness with no known preventive measures. Excess to insulin therapy can lead to hypoglycemia with neuro-glycogenic symptoms such as shakiness, nausea, sweating, irritability, fatigue, excessive thirst or hunger, weakness, seizure, and coma. Severe Hypoglycemia (SH) is also considered a most aversive event since it may put patients at risk for injury and death, which matches the criteria of a traumatic event. SH has a ranging prevalence of 20%, which makes it a primary medical Issue. One of the results of SH is an intense emotional fear reaction resembling the form of post-traumatic stress symptoms (PTS), causing many patients to avoid insulin therapy and social activities in order to avoid the possibility of hypoglycemia. As a result, they are at risk for irreversible health deterioration and medical complications. Fear of Hypoglycemia (FOH) is, therefore, a major disturbance for T1DM patients. FOH differs from prevalent post-traumatic stress reactions to other forms of traumatic events since the threat to life continuously exists in the patient's body. That is, it is highly probable that orthodox interventions may not be sufficient for helping patients after SH to regain healthy social function and proper medical treatment. Accordingly, the current presentation will demonstrate the results of a study conducted among T1DM patients after SH. The study was designed in two stages. First, a preliminary qualitative phenomenological study among ten patients after SH was conducted. Analysis revealed that after SH, patients confuse between stress symptoms and Hypoglycemia symptoms, divide life before and after the event, report a constant sense of fear, a loss of freedom, a significant decrease in social functioning, a catastrophic thinking pattern, a dichotomous split between the self and the body, and internalization of illness identity, a loss of internal locus of control, a damaged self-representation, and severe loneliness for never being understood by others. The second stage was a two steps study of intervention among five patients after SH. The first part of the intervention included three months of therapeutic 3rd wave CBT therapy. The contents of the therapeutic process were: acceptance of fear and tolerance to stress; cognitive de-fusion combined with emotional self-regulation; the adoption of an active position relying on personal values; and self-compassion. Then, the intervention included a one-week practical real-time 24/7 support by trained medical personnel, alongside a gradual exposure to increased insulin therapy in a protected environment. The results of the intervention are a decrease in stress symptoms, increased social functioning, increased well-being, and decreased avoidance of medical treatment. The presentation will discuss the unique emotional state of T1DM patients after SH. Then, the presentation will discuss the effectiveness of the intervention for patients with chronic conditions after a traumatic event. The presentation will make evident the unique situation of illness-related PTSD. The presentation will also demonstrate the requirement for multi-professional collaboration between social work and medical care for populations with chronic medical conditions. Limitations of the study and recommendations for further research will be discussed.

Keywords: type 1 diabetes, chronic illness, post-traumatic stress, illness-related PTSD

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Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

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Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

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Authors: Irina Mizeva, Elena Potapova, Viktor Dremin, Mikhail Mezentsev, Valeri Shupletsov

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The local tissue perfusion is regulated by the microvascular tone which is under the control of a number of physiological mechanisms. Laser Doppler flowmetry (LDF) together with wavelet analyses is the most commonly used technique to study the regulatory mechanisms of cutaneous microcirculation. External factors such as temperature, local pressure of the probe on the skin, etc. influence on the blood flow characteristics and are used as physiological tests to evaluate microvascular regulatory mechanisms. Local probe pressure influences on the microcirculation parameters measured by optical methods: diffuse reflectance spectroscopy, fluorescence spectroscopy, and LDF. Therefore, further study of probe pressure effects can be useful to improve the reliability of optical measurement. During pressure tests variation of the mean perfusion measured by means of LDF usually is estimated. An additional information concerning the physiological mechanisms of the vascular tone regulation system in response to local pressure can be obtained using spectral analyses of LDF samples. The aim of the present work was to develop protocol and algorithm of data processing appropriate for study physiological response to the local pressure test. Involving 6 subjects (20±2 years) and providing 5 measurements for every subject we estimated intersubject and-inter group variability of response of both averaged and oscillating parts of the LDF sample on external surface pressure. The final purpose of the work was to find special features which further can be used in wider clinic studies. The cutaneous perfusion measurements were carried out by LAKK-02 (SPE LAZMA Ltd., Russia), the skin loading was provided by the originally designed device which allows one to distribute the pressure around the LDF probe. The probe was installed on the dorsal part of the distal finger of the index figure. We collected measurements continuously for one hour and varied loading from 0 to 180mmHg stepwise with a step duration of 10 minutes. Further, we post-processed the samples using the wavelet transform and traced the energy of oscillations in five frequency bands over time. Weak loading leads to pressure-induced vasodilation, so one should take into account that the perfusion measured under pressure conditions will be overestimated. On the other hand, we revealed a decrease in endothelial associated fluctuations. Further loading (88 mmHg) induces amplification of pulsations in all frequency bands. We assume that such loading leads to a higher number of closed capillaries, higher input of arterioles in the LDF signal and as a consequence more vivid oscillations which mainly are formed in arterioles. External pressure higher than 144 mmHg leads to the decrease of oscillating components, after removing the loading very rapid restore of the tissue perfusion takes place. In this work, we have demonstrated that local skin loading influence on the microcirculation parameters measured by optic technique; this should be taken into account while developing portable electronic devices. The proposed protocol of local loading allows one to evaluate PIV as far as to trace dynamic of blood flow oscillations. This study was supported by the Russian Science Foundation under project N 18-15-00201.

Keywords: blood microcirculation, laser Doppler flowmetry, pressure-induced vasodilation, wavelet analyses blood

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Authors: Xu Qian

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This abstract explores the importance of building resilience within families and offers evidence-based strategies for promoting positive parenting and enhancing emotional well-being. It emphasizes the role of effective communication, conflict resolution, and fostering a supportive environment to strengthen family bonds and promote healthy child development. Introduction: The well-being and resilience of families play a crucial role in fostering healthy child development and promoting overall emotional well-being. This abstract highlights the significance of nurturing resilient families and provides evidence-based strategies for positive parenting. By focusing on effective communication, conflict resolution, and creating a supportive environment, families can strengthen their bonds and enhance emotional well-being for both parents and children. Methods: This abstract draws upon a comprehensive review of existing research and literature on resilient families, positive parenting, and emotional well-being. The selected studies employ various methodologies, including surveys, interviews, and longitudinal observations, to investigate the factors contributing to family resilience and the strategies that promote positive parenting practices. The findings from these studies serve as the foundation for the strategies discussed in this abstract. Results: The results of the reviewed studies demonstrate that effective communication within families is a key factor in building resilience and promoting emotional well-being. Open and honest communication allows family members to express their thoughts, feelings, and concerns, fostering trust and understanding. Conflict resolution skills, such as active listening, compromise, and problem-solving, are vital in managing conflicts constructively and preventing negative consequences on family dynamics and children's well-being. Creating a supportive environment that nurtures emotional well-being is another critical aspect of promoting resilient families. This includes providing emotional support, setting clear boundaries, and promoting positive discipline strategies. Research indicates that consistent and responsive parenting approaches contribute to improved self-regulation skills, emotional intelligence, and overall mental health in children. Discussion: The discussion centers on the implications of these findings for promoting positive parenting and emotional well-being. It emphasizes the need for parents to prioritize self-care and seek support when facing challenges. Parental well-being directly influences the quality of parenting and the overall family environment. By attending to their own emotional needs, parents can better meet the needs of their children and create a nurturing atmosphere. Furthermore, the importance of fostering resilience in children is highlighted. Resilient children are better equipped to cope with adversity, adapt to change, and thrive in challenging circumstances. By cultivating resilience through supportive relationships, encouragement of independence, and providing opportunities for growth, parents can foster their children's ability to bounce back from setbacks and develop essential life skills. Conclusion: In conclusion, nurturing resilient families is crucial for positive parenting and enhancing emotional well-being. This abstract presents evidence-based strategies that emphasize effective communication, conflict resolution, and creating a supportive environment. By implementing these strategies, parents can strengthen family bonds, promote healthy child development, and enhance overall family resilience. Investing in resilient families not only benefits individual family members but also contributes to the well-being of the broader community.

Keywords: childrearing families, family education, children's mental health, positive parenting, emotional health

Procedia PDF Downloads 87