Search results for: face detection
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Paper Count: 5827

Search results for: face detection

37 Role of Toll Like Receptor-2 in Female Genital Tuberculosis Disease Infection and Its Severity

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

Abstract:

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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36 Preparation and Struggle of Two Generations for Future Care: A Study of Intergenerational Care Planning among Mainland Immigrant Ageing Families in Hong Kong

Authors: Xue Bai, Ranran He, Chang Liu

Abstract:

Care planning before the onset of intensive care needs can benefit older adults’ psychological well-being and increases families’ ability to manage caregiving crises and cope with care transitions. Effective care planning requires collaborative ‘team-work’ in families. However, future care planning has not been substantially examined in intergenerational or family contexts, let alone among immigrant families who have to face particular challenges in parental caregiving. From a family systems perspective, this study intends to explore the extent, processes, and contents of intergenerational care planning of Mainland immigrant ageing families in Hong Kong and to examine the intergenerational congruence and discrepancies in the care planning process. Adopting a qualitative research design, semi-structured in-depth interviews were conducted with 17 adult child-older parent pairs and another 33 adult children. In total, 50 adult children who migrated to Hong Kong after the age of 18 with more than three years’ work experience in Hong Kong had at least one parent aged over 55 years old who was not a Hong Kong resident and considered his/herself as the primary caregiver of the parent were recruited. Seventeen ageing parents of the recruited adult children were invited for dyadic interviews. Scarcity of caregiving resources in the context of cross-border migration, intergenerational discrepancies in care planning stages, both generations’ struggle and ambivalence toward filial care, intergenerational transmission of care values, and facilitating role of accumulated family capital in care preparation were primary themes concluded from participants’ narratives. Compared with ageing parents, immigrant adult children generally displayed lower levels of care planning. Although with a strong awareness of parents’ future care needs, few adult children were found engaged in concrete planning activities. This is largely due to their uncertainties toward future life and career, huge work and living pressure, the relatively good health status of their parents, and restrictions of public welfare policies in the receiving society. By contrast, children’s cross-border migration encouraged ageing parents to have early and clear preparation for future care. Ageing parents mostly expressed low filial care expectations when realizing the scarcity of family caregiving resources in the cross-border context. Even though they prefer in-person support from children, most of them prepare themselves for independent ageing to prioritize the next generation’s needs or choose to utilize paid services, welfare systems, friend networks, or extended family networks in their sending society. Adult children were frequently found caught in the dilemma of desiring to provide high quality and in-person support for their parents but lacking sufficient resources. Notably, a salient pattern of intergenerational transmission in terms of family and care values and ideal care arrangement emerged from intergenerational care preparation. Moreover, the positive role of accumulated family capital generated by a reunion in care preparation and joint decision-making were also identified. The findings of the current study will enhance professionals’ and service providers’ awareness of intergenerational care planning in cross-border migration contexts, inform services to alleviate unpreparedness for elderly care and intergenerational discrepancies concerning care arrangements and broaden family services to encompass intergenerational care planning interventions. Acknowledgment: This study is supported by a General Research Grant from the Research Grants Council of the HKSAR, China (Project Number: 15603818).

Keywords: intergenerational care planning, mainland immigrants in Hong Kong, migrant family, older adults

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35 Differential Expression Profile Analysis of DNA Repair Genes in Mycobacterium Leprae by qPCR

Authors: Mukul Sharma, Madhusmita Das, Sundeep Chaitanya Vedithi

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Leprosy is a chronic human disease caused by Mycobacterium leprae, that cannot be cultured in vitro. Though treatable with multidrug therapy (MDT), recently, bacteria reported resistance to multiple antibiotics. Targeting DNA replication and repair pathways can serve as the foundation of developing new anti-leprosy drugs. Due to the absence of an axenic culture medium for the propagation of M. leprae, studying cellular processes, especially those belonging to DNA repair pathways, is challenging. Genomic understanding of M. Leprae harbors several protein-coding genes with no previously assigned function known as 'hypothetical proteins'. Here, we report identification and expression of known and hypothetical DNA repair genes from a human skin biopsy and mouse footpads that are involved in base excision repair, direct reversal repair, and SOS response. Initially, a bioinformatics approach was employed based on sequence similarity, identification of known protein domains to screen the hypothetical proteins in the genome of M. leprae, that are potentially related to DNA repair mechanisms. Before testing on clinical samples, pure stocks of bacterial reference DNA of M. leprae (NHDP63 strain) was used to construct standard graphs to validate and identify lower detection limit in the qPCR experiments. Primers were designed to amplify the respective transcripts, and PCR products of the predicted size were obtained. Later, excisional skin biopsies of newly diagnosed untreated, treated, and drug resistance leprosy cases from SIHR & LC hospital, Vellore, India were taken for the extraction of RNA. To determine the presence of the predicted transcripts, cDNA was generated from M. leprae mRNA isolated from clinically confirmed leprosy skin biopsy specimen across all the study groups. Melting curve analysis was performed to determine the integrity of the amplification and to rule out primer‑dimer formation. The Ct values obtained from qPCR were fitted to standard curve to determine transcript copy number. Same procedure was applied for M. leprae extracted after processing a footpad of nude mice of drug sensitive and drug resistant strains. 16S rRNA was used as positive control. Of all the 16 genes involved in BER, DR, and SOS, differential expression pattern of the genes was observed in terms of Ct values when compared to human samples; this was because of the different host and its immune response. However, no drastic variation in gene expression levels was observed in human samples except the nth gene. The higher expression of nth gene could be because of the mutations that may be associated with sequence diversity and drug resistance which suggests an important role in the repair mechanism and remains to be explored. In both human and mouse samples, SOS system – lexA and RecA, and BER genes AlkB and Ogt were expressing efficiently to deal with possible DNA damage. Together, the results of the present study suggest that DNA repair genes are constitutively expressed and may provide a reference for molecular diagnosis, therapeutic target selection, determination of treatment and prognostic judgment in M. leprae pathogenesis.

Keywords: DNA repair, human biopsy, hypothetical proteins, mouse footpads, Mycobacterium leprae, qPCR

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34 Implementation of Cord- Blood Derived Stem Cells in the Regeneration of Two Experimental Models: Carbon Tetrachloride and S. Mansoni Induced Liver Fibrosis

Authors: Manal M. Kame, Zeinab A. Demerdash, Hanan G. El-Baz, Salwa M. Hassan, Faten M. Salah, Wafaa Mansour, Olfat Hammam

Abstract:

Cord blood (CB) derived Unrestricted Somatic Stem Cells (USSCs) with their multipotentiality hold great promise in liver regeneration. This work aims at evaluation of the therapeutic potentiality of USSCs in two experimental models of chronic liver injury induced either by S. mansoni infection in balb/c mice or CCL4 injection in hamsters. Isolation, propagation, and characterization of USSCs from CB samples were performed. USSCs were induced to differentiate into osteoblasts, adipocytes and hepatocyte-like cells. Cells of the third passage were transplanted in two models of liver fibrosis: (1) Twenty hamsters were induced to liver fibrosis by repeated i. p. injection of 100 μl CCl4 /hamster for 8 weeks. This model was designed as; 10 hamsters with liver fibrosis and treated with i.h. injection of 3x106 USSCs (USSCs transplanted group), 10 hamsters with liver fibrosis (pathological control group), and 10 hamsters with healthy livers (normal control group). (2) Murine chronics S.mansoni model: twenty mice were induced to liver fibrosis with S. mansoni ceracariae (60 cercariae/ mouse) using the tail immersion method and left for 12 weeks. This model was designed as; 10 mice with liver fibrosis were transplanted with i. v. injection of 1×106 USCCs (USSCs transplanted group). Other 2 groups were designed as in hamsters model. Animals were sacrificed 12 weeks after USSCs transplantation, and their liver sections were examined for detection of human hepatocyte-like cells by immunohistochemistry staining. Moreover, liver sections were examined for fibrosis level, and fibrotic indices were calculated. Sera of sacrificed animals were tested for liver functions. CB USSCs, with fibroblast-like morphology, expressed high levels of CD44, CD90, CD73 and CD105 and were negative for CD34, CD45, and HLA-DR. USSCs showed high expression of transcripts for Oct4 and Sox2 and were in vitro differentiated into osteoblasts, adipocytes. In both animal models, in vitro induced hepatocyte-like cells were confirmed by cytoplasmic expression of glycogen, alpha-fetoprotein, and cytokeratin18. Livers of USSCs transplanted group showed engraftment with human hepatocyte-like cells as proved by cytoplasmic expression of human alpha-fetoprotein, cytokeratin18, and OV6. In addition, livers of this group showed less fibrosis than the pathological control group. Liver functions in the form of serum AST & ALT level and serum total bilirubin level were significantly lowered in USSCs transplanted group than pathological control group (p < 0.001). Moreover, the fibrotic index was significantly lower (p< 0.001) in USSCs transplanted group than pathological control group. In addition liver sections, of i. v. injection of 1×106 USCCs of mice, stained with either H&E or sirius red showed diminished granuloma size and a relative decrease in hepatic fibrosis. Our experimental liver fibrosis models transplanted with CB-USSCs showed liver engraftment with human hepatocyte-like cells as well as signs of liver regeneration in the form of improvement in liver function assays and fibrosis level. These data provide hope that human CB- derived USSCs are introduced as multipotent stem cells with great potentiality in regenerative medicine & strengthens the concept of cellular therapy for the treatment of liver fibrosis.

Keywords: cord blood, liver fibrosis, stem cells, transplantation

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33 Early Predictive Signs for Kasai Procedure Success

Authors: Medan Isaeva, Anna Degtyareva

Abstract:

Context: Biliary atresia is a common reason for liver transplants in children, and the Kasai procedure can potentially be successful in avoiding the need for transplantation. However, it is important to identify factors that influence surgical outcomes in order to optimize treatment and improve patient outcomes. Research aim: The aim of this study was to develop prognostic models to assess the outcomes of the Kasai procedure in children with biliary atresia. Methodology: This retrospective study analyzed data from 166 children with biliary atresia who underwent the Kasai procedure between 2002 and 2021. The effectiveness of the operation was assessed based on specific criteria, including post-operative stool color, jaundice reduction, and bilirubin levels. The study involved a comparative analysis of various parameters, such as gestational age, birth weight, age at operation, physical development, liver and spleen sizes, and laboratory values including bilirubin, ALT, AST, and others, measured pre- and post-operation. Ultrasonographic evaluations were also conducted pre-operation, assessing the hepatobiliary system and related quantitative parameters. The study was carried out by two experienced specialists in pediatric hepatology. Comparative analysis and multifactorial logistic regression were used as the primary statistical methods. Findings: The study identified several statistically significant predictors of a successful Kasai procedure, including the presence of the gallbladder and levels of cholesterol and direct bilirubin post-operation. A detectable gallbladder was associated with a higher probability of surgical success, while elevated post-operative cholesterol and direct bilirubin levels were indicative of a reduced chance of positive outcomes. Theoretical importance: The findings of this study contribute to the optimization of treatment strategies for children with biliary atresia undergoing the Kasai procedure. By identifying early predictive signs of success, clinicians can modify treatment plans and manage patient care more effectively and proactively. Data collection and analysis procedures: Data for this analysis were obtained from the health records of patients who received the Kasai procedure. Comparative analysis and multifactorial logistic regression were employed to analyze the data and identify significant predictors. Question addressed: The study addressed the question of identifying predictive factors for the success of the Kasai procedure in children with biliary atresia. Conclusion: The developed prognostic models serve as valuable tools for early detection of patients who are less likely to benefit from the Kasai procedure. This enables clinicians to modify treatment plans and manage patient care more effectively and proactively. Potential limitations of the study: The study has several limitations. Its retrospective nature may introduce biases and inconsistencies in data collection. Being single centered, the results might not be generalizable to wider populations due to variations in surgical and postoperative practices. Also, other potential influencing factors beyond the clinical, laboratory, and ultrasonographic parameters considered in this study were not explored, which could affect the outcomes of the Kasai operation. Future studies could benefit from including a broader range of factors.

Keywords: biliary atresia, kasai operation, prognostic model, native liver survival

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32 Use of Machine Learning Algorithms to Pediatric MR Images for Tumor Classification

Authors: I. Stathopoulos, V. Syrgiamiotis, E. Karavasilis, A. Ploussi, I. Nikas, C. Hatzigiorgi, K. Platoni, E. P. Efstathopoulos

Abstract:

Introduction: Brain and central nervous system (CNS) tumors form the second most common group of cancer in children, accounting for 30% of all childhood cancers. MRI is the key imaging technique used for the visualization and management of pediatric brain tumors. Initial characterization of tumors from MRI scans is usually performed via a radiologist’s visual assessment. However, different brain tumor types do not always demonstrate clear differences in visual appearance. Using only conventional MRI to provide a definite diagnosis could potentially lead to inaccurate results, and so histopathological examination of biopsy samples is currently considered to be the gold standard for obtaining definite diagnoses. Machine learning is defined as the study of computational algorithms that can use, complex or not, mathematical relationships and patterns from empirical and scientific data to make reliable decisions. Concerning the above, machine learning techniques could provide effective and accurate ways to automate and speed up the analysis and diagnosis for medical images. Machine learning applications in radiology are or could potentially be useful in practice for medical image segmentation and registration, computer-aided detection and diagnosis systems for CT, MR or radiography images and functional MR (fMRI) images for brain activity analysis and neurological disease diagnosis. Purpose: The objective of this study is to provide an automated tool, which may assist in the imaging evaluation and classification of brain neoplasms in pediatric patients by determining the glioma type, grade and differentiating between different brain tissue types. Moreover, a future purpose is to present an alternative way of quick and accurate diagnosis in order to save time and resources in the daily medical workflow. Materials and Methods: A cohort, of 80 pediatric patients with a diagnosis of posterior fossa tumor, was used: 20 ependymomas, 20 astrocytomas, 20 medulloblastomas and 20 healthy children. The MR sequences used, for every single patient, were the following: axial T1-weighted (T1), axial T2-weighted (T2), FluidAttenuated Inversion Recovery (FLAIR), axial diffusion weighted images (DWI), axial contrast-enhanced T1-weighted (T1ce). From every sequence only a principal slice was used that manually traced by two expert radiologists. Image acquisition was carried out on a GE HDxt 1.5-T scanner. The images were preprocessed following a number of steps including noise reduction, bias-field correction, thresholding, coregistration of all sequences (T1, T2, T1ce, FLAIR, DWI), skull stripping, and histogram matching. A large number of features for investigation were chosen, which included age, tumor shape characteristics, image intensity characteristics and texture features. After selecting the features for achieving the highest accuracy using the least number of variables, four machine learning classification algorithms were used: k-Nearest Neighbour, Support-Vector Machines, C4.5 Decision Tree and Convolutional Neural Network. The machine learning schemes and the image analysis are implemented in the WEKA platform and MatLab platform respectively. Results-Conclusions: The results and the accuracy of images classification for each type of glioma by the four different algorithms are still on process.

Keywords: image classification, machine learning algorithms, pediatric MRI, pediatric oncology

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31 Fine Characterization of Glucose Modified Human Serum Albumin by Different Biophysical and Biochemical Techniques at a Range

Authors: Neelofar, Khursheed Alam, Jamal Ahmad

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Protein modification in diabetes mellitus may lead to early glycation products (EGPs) or amadori product as well as advanced glycation end products (AGEs). Early glycation involves the reaction of glucose with N-terminal and lysyl side chain amino groups to form Schiff’s base which undergoes rearrangements to form more stable early glycation product known as Amadori product. After Amadori, the reactions become more complicated leading to the formation of advanced glycation end products (AGEs) that interact with various AGE receptors, thereby playing an important role in the long-term complications of diabetes. Millard reaction or nonenzymatic glycation reaction accelerate in diabetes due to hyperglycation and alter serum protein’s structure, their normal functions that lead micro and macro vascular complications in diabetic patients. In this study, Human Serum Albumin (HSA) with a constant concentration was incubated with different concentrations of glucose at 370C for a week. At 4th day, Amadori product was formed that was confirmed by colorimetric method NBT assay and TBA assay which both are authenticate early glycation product. Conformational changes in native as well as all samples of Amadori albumin with different concentrations of glucose were investigated by various biophysical and biochemical techniques. Main biophysical techniques hyperchromacity, quenching of fluorescence intensity, FTIR, CD and SDS-PAGE were used. Further conformational changes were observed by biochemical assays mainly HMF formation, fructoseamine, reduction of fructoseamine with NaBH4, carbonyl content estimation, lysine and arginine residues estimation, ANS binding property and thiol group estimation. This study find structural and biochemical changes in Amadori modified HSA with normal to hyperchronic range of glucose with respect to native HSA. When glucose concentration was increased from normal to chronic range biochemical and structural changes also increased. Highest alteration in secondary and tertiary structure and conformation in glycated HSA was observed at the hyperchronic concentration (75mM) of glucose. Although it has been found that Amadori modified proteins is also involved in secondary complications of diabetes as AGEs but very few studies have been done to analyze the conformational changes in Amadori modified proteins due to early glycation. Most of the studies were found on the structural changes in Amadori protein at a particular glucose concentration but no study was found to compare the biophysical and biochemical changes in HSA due to early glycation with a range of glucose concentration at a constant incubation time. So this study provide the information about the biochemical and biophysical changes occur in Amadori modified albumin at a range of glucose normal to chronic in diabetes. Although many implicates currently in use i.e. glycaemic control, insulin treatment and other chemical therapies that can control many aspects of diabetes. However, even with intensive use of current antidiabetic agents more than 50 % of diabetic patient’s type 2 suffers poor glycaemic control and 18 % develop serious complications within six years of diagnosis. Experimental evidence related to diabetes suggests that preventing the nonenzymatic glycation of relevant proteins or blocking their biological effects might beneficially influence the evolution of vascular complications in diabetic patients or quantization of amadori adduct of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes. So this research work may be helpful for the same.

Keywords: diabetes mellitus, glycation, albumin, amadori, biophysical and biochemical techniques

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30 Musictherapy and Gardentherapy: A Systemic Approach for the Life Quality of the PsychoPhysical Disability

Authors: Adriana De Serio, Donato Forenza

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Aims. In this experimental research the Authors present the methodological plan “Musictherapy and Gardentherapy” that they created interconnected with the garden landscape ecosystems and aimed at PsychoPhysical Disability (MusGarPPhyD). In the context of the environmental education aimed at spreading the landscape culture and its values, it’s necessary to develop a solid perception of the environment sustainability to implement a multidimensional approach that pays attention to the conservation and enhancement of gardens and natural environments. The result is an improvement in the life quality also in compliance with the objectives of the European Agenda 2030. The MusGarPPhyD can help professionals such as musictherapists and environmental and landscape researchers strengthen subjects' motivation to learn to deal with the psychophysical discomfort associated with disability and to cope with the distress and the psychological fragility and the loneliness and the social seclusion and to promote productive social relationships. Materials and Methods. The MusGarPPhyD was implemented in multiple spaces. The musictherapy treatments took place first inside residential therapeutic centres and then in the garden landscape ecosystem. Patients: twenty, set in two groups. Weekly-sessions (50’) for three months. Methodological phases: - Phase P1. MusicTherapy treatments for each group in the indoor spaces. - Phase P2. MusicTherapy sessions inside the gardens. After each Phase, P1 and P2: - a Questionnaire for each patient (ten items / liking-indices) was administrated at t0 time, during the treatment and at tn time at the end of the treatment. - Monitoring of patients' behavioral responses through assessment scales, matrix, table and graph system. MusicTherapy methodology: pazient Sonorous-Musical Anamnesis, Musictherapy Assessment Document, Observation Protocols, Bodily-Environmental-Rhythmical-Sonorous-Vocal-Energy production first indoors and then outside, sonorous-musical instruments and edible instruments made by the Author/musictherapist with some foods; Administration of Patient-Environment-Music Index at time to and tn, to estimate the patient’s behavior evolution, Musictherapeutic Advancement Index. Results. The MusGarPPhyD can strengthen the individual sense of identity and improve the psychophysical skills and the resilience to face and to overcome the difficulties caused by the congenital /acquired disability. The multi-sensory perceptions deriving from contact with the plants in the gardens improve the psychological well-being and regulate the physiological parameters such as blood pressure, cardiac and respiratory rhythm, reducing the cholesterol levels. The secretions of the peptide hormones endorphins and the endogenous opioids enkephalins increase and bring a state of patient’s tranquillity and a better mood. The subjects showed a preference for musictherapy treatments within a setting made up of gardens and peculiar landscape systems. This resulted in greater health benefits. Conclusions. The MusGarPPhyD contributes to reduce psychophysical tensions, anxiety, depression and stress, facilitating the connections between the cerebral hemispheres, thus also improving intellectual performances, self-confidence, motor skills and social interactions. Therefore it is necessary to design hospitals, rehabilitation centers, nursing homes, surrounded by gardens. Ecosystems of natural and urban parks and gardens create fascinating skyline and mosaics of landscapes rich in beauty and biodiversity. The MusGarPPhyD is useful for the health management promoting patient’s psychophysical activation, better mood/affective-tone and relastionships and contributing significantly to improving the life quality.

Keywords: musictherapy, gardentherapy, disability, life quality

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29 Raman Spectral Fingerprints of Healthy and Cancerous Human Colorectal Tissues

Authors: Maria Karnachoriti, Ellas Spyratou, Dimitrios Lykidis, Maria Lambropoulou, Yiannis S. Raptis, Ioannis Seimenis, Efstathios P. Efstathopoulos, Athanassios G. Kontos

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Colorectal cancer is the third most common cancer diagnosed in Europe, according to the latest incidence data provided by the World Health Organization (WHO), and early diagnosis has proved to be the key in reducing cancer-related mortality. In cases where surgical interventions are required for cancer treatment, the accurate discrimination between healthy and cancerous tissues is critical for the postoperative care of the patient. The current study focuses on the ex vivo handling of surgically excised colorectal specimens and the acquisition of their spectral fingerprints using Raman spectroscopy. Acquired data were analyzed in an effort to discriminate, in microscopic scale, between healthy and malignant margins. Raman spectroscopy is a spectroscopic technique with high detection sensitivity and spatial resolution of few micrometers. The spectral fingerprint which is produced during laser-tissue interaction is unique and characterizes the biostructure and its inflammatory or cancer state. Numerous published studies have demonstrated the potential of the technique as a tool for the discrimination between healthy and malignant tissues/cells either ex vivo or in vivo. However, the handling of the excised human specimens and the Raman measurement conditions remain challenging, unavoidably affecting measurement reliability and repeatability, as well as the technique’s overall accuracy and sensitivity. Therefore, tissue handling has to be optimized and standardized to ensure preservation of cell integrity and hydration level. Various strategies have been implemented in the past, including the use of balanced salt solutions, small humidifiers or pump-reservoir-pipette systems. In the current study, human colorectal specimens of 10X5 mm were collected from 5 patients up to now who underwent open surgery for colorectal cancer. A novel, non-toxic zinc-based fixative (Z7) was used for tissue preservation. Z7 demonstrates excellent protein preservation and protection against tissue autolysis. Micro-Raman spectra were recorded with a Renishaw Invia spectrometer from successive random 2 micrometers spots upon excitation at 785 nm to decrease fluorescent background and secure avoidance of tissue photodegradation. A temperature-controlled approach was adopted to stabilize the tissue at 2 °C, thus minimizing dehydration effects and consequent focus drift during measurement. A broad spectral range, 500-3200 cm-1,was covered with five consecutive full scans that lasted for 20 minutes in total. The average spectra were used for least square fitting analysis of the Raman modes.Subtle Raman differences were observed between normal and cancerous colorectal tissues mainly in the intensities of the 1556 cm-1 and 1628 cm-1 Raman modes which correspond to v(C=C) vibrations in porphyrins, as well as in the range of 2800-3000 cm-1 due to CH2 stretching of lipids and CH3 stretching of proteins. Raman spectra evaluation was supported by histological findings from twin specimens. This study demonstrates that Raman spectroscopy may constitute a promising tool for real-time verification of clear margins in colorectal cancer open surgery.

Keywords: colorectal cancer, Raman spectroscopy, malignant margins, spectral fingerprints

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28 Using the UK as a Case Study to Assess the Current State of Large Woody Debris Restoration as a Tool for Improving the Ecological Status of Natural Watercourses Globally

Authors: Isabelle Barrett

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Natural watercourses provide a range of vital ecosystem services, notably freshwater provision. They also offer highly heterogeneous habitat which supports an extreme diversity of aquatic life. Exploitation of rivers, changing land use and flood prevention measures have led to habitat degradation and subsequent biodiversity loss; indeed, freshwater species currently face a disproportionate rate of extinction compared to their terrestrial and marine counterparts. Large woody debris (LWD) encompasses the trees, large branches and logs which fall into watercourses, and is responsible for important habitat characteristics. Historically, natural LWD has been removed from streams under the assumption that it is not aesthetically pleasing and is thus ecologically unfavourable, despite extensive evidence contradicting this. Restoration efforts aim to replace lost LWD in order to reinstate habitat heterogeneity. This paper aims to assess the current state of such restoration schemes for improving fluvial ecological health in the UK. A detailed review of the scientific literature was conducted alongside a meta-analysis of 25 UK-based projects involving LWD restoration. Projects were chosen for which sufficient information was attainable for analysis, covering a broad range of budgets and scales. The most effective strategies for river restoration encompass ecological success, stakeholder engagement and scientific advancement, however few projects surveyed showed sensitivity to all three; for example, only 32% of projects stated biological aims. Focus tended to be on stakeholder engagement and public approval, since this is often a key funding driver. Consequently, there is a tendency to focus on the aesthetic outcomes of a project, however physical habitat restoration does not necessarily lead to direct biodiversity increases. This highlights the significance of rivers as highly heterogeneous environments with multiple interlinked processes, and emphasises a need for a stronger scientific presence in project planning. Poor scientific rigour means monitoring is often lacking, with varying, if any, definitions of success which are rarely pre-determined. A tendency to overlook negative or neutral results was apparent, with unjustified focus often put on qualitative results. The temporal scale of monitoring is typically inadequate to facilitate scientific conclusions, with only 20% of projects surveyed reporting any pre-restoration monitoring. Furthermore, monitoring is often limited to a few variables, with biotic monitoring often fish-focussed. Due to their longer life cycles and dispersal capability, fish are usually poor indicators of environmental change, making it difficult to attribute any changes in ecological health to restoration efforts. Although the potential impact of LWD restoration may be positive, this method of restoration could simply be making short-term, small-scale improvements; without addressing the underlying symptoms of degradation, for example water quality, the issue cannot be fully resolved. Promotion of standardised monitoring for LWD projects could help establish a deeper understanding of the ecology surrounding the practice, supporting movement towards adaptive management in which scientific evidence feeds back to practitioners, enabling the design of more efficient projects with greater ecological success. By highlighting LWD, this study hopes to address the difficulties faced within river management, and emphasise the need for a more holistic international and inter-institutional approach to tackling problems associated with degradation.

Keywords: biological monitoring, ecological health, large woody debris, river management, river restoration

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27 Research Cooperation between of Ukraine in Terms of Food Chain Safety Control in the Frame of MICRORISK Project

Authors: Kinga Wieczorek, Elzbieta Kukier, Remigiusz Pomykala, Beata Lachtara, Renata Szewczyk, Krzysztof Kwiatek, Jacek Osek

Abstract:

The MICRORISK project (Research cooperation in assessment of microbiological hazard and risk in the food chain) was funded by the European Commission under the FP7 PEOPLE 2012 IRSES call within the International Research Staff Exchange Scheme of Marie Curie Action and realized during years from 2014 to 2015. The main aim of the project was to establish a cooperation between the European Union (EU) and the third State in the area important from the public health point of view. The following organizations have been engaged in the activity: National Veterinary Research Institute (NVRI) in Pulawy, Poland (coordinator), French Agency for Food, Environmental and Occupational Health & Safety (ANSES) in Maisons Alfort, France, National Scientific Center Institute of Experimental and Clinical Veterinary Medicine (NSC IECVM), Kharkov and State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise (SSRILDVSE) Kijev Ukraine. The results of the project showed that Ukraine used microbiological criteria in accordance with Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs. Compliance concerns both the criteria applicable at the stage of food safety (retail trade), as well as evaluation criteria and process hygiene in food production. In this case, the Ukrainian legislation also provides application of the criteria that do not have counterparts in the food law of the European Union, and are based on the provisions of Ukrainian law. Partial coherence of the Ukrainian and EU legal requirements in terms of microbiological criteria for food and feed concerns microbiological parameters such as total plate count, coliforms, coagulase-positive Staphylococcus spp., including S. aureus. Analysis of laboratory methods used for microbiological hazards control in food production chain has shown that most methods used in the EU are well-known by Ukrainian partners, and many of them are routinely applied as the only standards in the laboratory practice or simultaneously used with Ukrainian methods. The area without any legislation, where the EU regulation and analytical methods should be implemented is the area of Shiga toxin producing E. coli, including E. coli O157 and staphylococcal enterotoxin detection. During the project, the analysis of the existing Ukrainian and EU data concerning the prevalence of the most important food-borne pathogens on different stages of food production chain was performed. Particularly, prevalence of Salmonella spp., Campylobacter spp., L. monocytogenes as well as clostridia was examined. The analysis showed that poultry meat still appears to be the most important food-borne source of Campylobacter and Salmonella in the UE. On the other hand, L. monocytogenes were seldom detected above the legal safety limit (100 cfu/g) among the EU countries. Moreover, the analysis revealed the lack of comprehensive data regarding the prevalence of the most important food-borne pathogens in Ukraine. The results of the MICRORISK project are networking activities among researches originations participating in the tasks will help with a better recognition of each other regarding very important, from the public health point of view areas such as microbiological hazards in the food production chain and finally will help to improve food quality and safety for consumers.

Keywords: cooperation, European Union, food chain safety, food law, microbiological risk, Microrisk, Poland, Ukraine

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26 Clinico-pathological Study of Xeroderma Pigmentosa: A Case Series of Eight Cases

Authors: Kakali Roy, Sahana P. Raju, Subhra Dhar, Sandipan Dhar

Abstract:

Introduction: Xeroderma pigmentosa (XP) is a rare inherited (autosomal recessive) disease resulting from impairment in DNA repair that involves recognition and repair of ultraviolet radiation (UVR) induced DNA damage in the nucleotide excision repair pathway. Which results in increased photosensitivity, UVR induced damage to skin and eye, increased susceptibility of skin and ocular cancer, and progressive neurodegeneration in some patients. XP is present worldwide, with higher incidence in areas having frequent consanguinity. Being extremely rare, there is limited literature on XP and associated complications. Here, the clinico-pathological experience (spectrum of clinical presentation, histopathological findings of malignant skin lesions, and progression) of managing 8 cases of XP is presented. Methodology: A retrospective study was conducted in a pediatric tertiary care hospital in eastern India during a ten-year period from 2013 to 2022. A clinical diagnosis was made based on severe sun burn or premature photo-aging and/or onset of cutaneous malignancies at early age (1st decade) in background of consanguinity and autosomal recessive inheritance pattern in family. Results: The mean age of presentation was 1.2 years (range of 7month-3years), while three children presented during their infancy. Male to female ratio was 5:3, and all were born of consanguineous marriage. They presented with dermatological manifestations (100%) followed by ophthalmic (75%) and/or neurological symptoms (25%). Patients had normal skin at birth but soon developed extreme sensitivity to UVR in the form of exaggerated sun tanning, burning, and blistering on minimal sun exposure, followed by abnormal skin pigmentation like freckles and lentiginosis. Subsequently, over time there was progressive xerosis, atrophy, wrinkling, and poikiloderma. Six patients had varied degree of ocular involvement, while three of them had severe manifestation, including madarosis, tylosis, ectropion, Lagopthalmos, Pthysis bulbi, clouding and scarring of the cornea with complete or partial loss of vision, and ophthalmic malignancies. 50% (n=4) cases had skin and ocular pre-malignant (actinic keratosis) and malignant lesions, including melanoma and non melanoma skin cancer (NMSC) like squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) in their early childhood. One patient had simultaneous occurrence of multiple malignancies together (SCC, BCC, and melanoma). Subnormal intelligence was noticed as neurological feature, and none had sensory neural hearing loss, microcephaly, neuroregression, or neurdeficit. All the patients had been being managed by a multidisciplinary team of pediatricians, dermatologists, ophthalmologists, neurologists and psychiatrists. Conclusion: Although till date there is no complete cure for XP and the disease is ultimately fatal. But increased awareness, early diagnosis followed by persistent vigorous protection from UVR, and regular screening for early detection of malignancies along with psychological support can drastically improve patients’ quality of life and life expectancy. Further research is required on formulating optimal management of XP, specifically the role and possibilities of gene therapy in XP.

Keywords: childhood malignancies, dermato-pathological findings, eastern India, Xeroderma pigmentosa

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25 Optimum Irrigation System Management for Climate Resilient and Improved Productivity of Flood-based Livelihood Systems

Authors: Mara Getachew Zenebe, Luuk Fleskens, Abdu Obieda Ahmed

Abstract:

This paper seeks to advance our scientific understanding of optimizing flood utilization in regions impacted by climate change, with a focus on enhancing agricultural productivity through effective irrigation management. The study was conducted as part of a three-year (2021 to 2023) USAID-supported initiative aimed at promoting Economic Growth and Peace in the Gash Agricultural Scheme (GAS), situated in Sudan's water-stressed Eastern region. GAS is the country's largest flood-irrigated scheme, covering 100,800 hectares of cultivable land, with a potential to provide the food security needs of over a quarter of a million agro-pastoral community members. GAS relies on the Gash River, which sources its water from high-intensity rainfall events in the highlands of Ethiopia and Eritrea. However, climate change and variations in these highlands have led to increased variability in the Gash River's flow. The study conducted water balance analyses based on a ten-year dataset of the annual Gash River flow, irrigated area; as well as the evapotranspiration demand of the major sorghum crop. Data collection methods included field measurements, surveys, remote sensing, and CropWat modelling. The water balance assessment revealed that the existing three-year rotation-based irrigation system management, capping cultivated land at 33,000 hectares annually, is excessively risk-averse. While this system reduced conflicts among the agro-pastoral communities by consistently delivering on the land promised to be annually cultivated, it also increased GAS's vulnerability to flood damage due to several reasons. The irrigation efficiency over the past decade was approximately 30%, leaving significant unharnessed floodwater that caused damage to infrastructure and agricultural land. The three-year rotation resulted in inadequate infrastructural maintenance, given the destructive nature of floods. Additionally, it led to infrequent land tillage, allowing the encroachment of mesquite trees hindering major sorghum crop growth. Remote sensing data confirmed that mesquite trees have overtaken 70,000 hectares in the past two decades, rendering them unavailable for agriculture. The water balance analyses suggest shifting to a two-year rotation, covering approximately 50,000 hectares annually while maintaining risk aversion. This shift could boost GAS's annual sorghum production by two-thirds, exceeding 850,000 tons. The scheme's efficiency can be further enhanced through low-cost on-farm interventions. Currently, large irrigation plots that range from 420 to 756 hectares are irrigated with limited water distribution guidance, leading to uneven irrigation. As demonstrated through field trials, implementing internal longitudinal bunds and horizontal deflector bunds can increase adequately irrigated parts of the irrigation plots from 50% to 80% and thus nearly double the sorghum yield to 2 tons per hectare while reducing the irrigation duration from 30 days to a maximum of 17 days. Flow measurements in 2021 and 2022 confirmed that these changes sufficiently meet the sorghum crop's water requirements, even with a conservative 60% field application efficiency assumption. These insights and lessons from the GAS on enhancing agricultural resilience and sustainability in the face of climate change are relevant to flood-based livelihood systems globally.

Keywords: climate change, irrigation management and productivity, variable flood flows, water balance analysis

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24 Electronic Raman Scattering Calibration for Quantitative Surface-Enhanced Raman Spectroscopy and Improved Biostatistical Analysis

Authors: Wonil Nam, Xiang Ren, Inyoung Kim, Masoud Agah, Wei Zhou

Abstract:

Despite its ultrasensitive detection capability, surface-enhanced Raman spectroscopy (SERS) faces challenges as a quantitative biochemical analysis tool due to the significant dependence of local field intensity in hotspots on nanoscale geometric variations of plasmonic nanostructures. Therefore, despite enormous progress in plasmonic nanoengineering of high-performance SERS devices, it is still challenging to quantitatively correlate the measured SERS signals with the actual molecule concentrations at hotspots. A significant effort has been devoted to developing SERS calibration methods by introducing internal standards. It has been achieved by placing Raman tags at plasmonic hotspots. Raman tags undergo similar SERS enhancement at the same hotspots, and ratiometric SERS signals for analytes of interest can be generated with reduced dependence on geometrical variations. However, using Raman tags still faces challenges for real-world applications, including spatial competition between the analyte and tags in hotspots, spectral interference, laser-induced degradation/desorption due to plasmon-enhanced photochemical/photothermal effects. We show that electronic Raman scattering (ERS) signals from metallic nanostructures at hotspots can serve as the internal calibration standard to enable quantitative SERS analysis and improve biostatistical analysis. We perform SERS with Au-SiO₂ multilayered metal-insulator-metal nano laminated plasmonic nanostructures. Since the ERS signal is proportional to the volume density of electron-hole occupation in hotspots, the ERS signals exponentially increase when the wavenumber is approaching the zero value. By a long-pass filter, generally used in backscattered SERS configurations, to chop the ERS background continuum, we can observe an ERS pseudo-peak, IERS. Both ERS and SERS processes experience the |E|⁴ local enhancements during the excitation and inelastic scattering transitions. We calibrated IMRS of 10 μM Rhodamine 6G in solution by IERS. The results show that ERS calibration generates a new analytical value, ISERS/IERS, insensitive to variations from different hotspots and thus can quantitatively reflect the molecular concentration information. Given the calibration capability of ERS signals, we performed label-free SERS analysis of living biological systems using four different breast normal and cancer cell lines cultured on nano-laminated SERS devices. 2D Raman mapping over 100 μm × 100 μm, containing several cells, was conducted. The SERS spectra were subsequently analyzed by multivariate analysis using partial least square discriminant analysis. Remarkably, after ERS calibration, MCF-10A and MCF-7 cells are further separated while the two triple-negative breast cancer cells (MDA-MB-231 and HCC-1806) are more overlapped, in good agreement with the well-known cancer categorization regarding the degree of malignancy. To assess the strength of ERS calibration, we further carried out a drug efficacy study using MDA-MB-231 and different concentrations of anti-cancer drug paclitaxel (PTX). After ERS calibration, we can more clearly segregate the control/low-dosage groups (0 and 1.5 nM), the middle-dosage group (5 nM), and the group treated with half-maximal inhibitory concentration (IC50, 15 nM). Therefore, we envision that ERS calibrated SERS can find crucial opportunities in label-free molecular profiling of complicated biological systems.

Keywords: cancer cell drug efficacy, plasmonics, surface-enhanced Raman spectroscopy (SERS), SERS calibration

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23 Impacts of School-Wide Positive Behavioral Interventions and Supports on Student Academics, Behavior and Mental Health

Authors: Catherine Bradshaw

Abstract:

Educators often report difficulty managing behavior problems and other mental health concerns that students display at school. These concerns also interfere with the learning process and can create distraction for teachers and other students. As such, schools play an important role in both preventing and intervening with students who experience these types of challenges. A number of models have been proposed to serve as a framework for delivering prevention and early intervention services in schools. One such model is called Positive Behavioral Interventions and Supports (PBIS), which has been scaled-up to over 26,000 schools in the U.S. and many other countries worldwide. PBIS aims to improve a range of student outcomes through early detection of and intervention related to behavioral and mental health symptoms. PBIS blends and applies social learning, behavioral, and organizational theories to prevent disruptive behavior and enhance the school’s organizational health. PBIS focuses on creating and sustaining tier 1 (universal), tier 2 (selective), and tier 3 (individual) systems of support. Most schools using PBIS have focused on the core elements of the tier 1 supports, which includes the following critical features. The formation of a PBIS team within the school to lead implementation. Identification and training of a behavioral support ‘coach’, who serves as a on-site technical assistance provider. Many of the individuals identified to serve as a PBIS coach are also trained as a school psychologist or guidance counselor; coaches typically have prior PBIS experience and are trained to conduct functional behavioral assessments. The PBIS team also identifies a set of three to five positive behavioral expectations that are implemented for all students and by all staff school-wide (e.g., ‘be respectful, responsible, and ready to learn’); these expectations are posted in all settings across the school, including in the classroom, cafeteria, playground etc. All school staff define and teach the school-wide behavioral expectations to all students and review them regularly. Finally, PBIS schools develop or adopt a school-wide system to reward or reinforce students who demonstrate those 3-5 positive behavioral expectations. Staff and administrators create an agreed upon system for responding to behavioral violations that include definitions about what constitutes a classroom-managed vs. an office-managed discipline problem. Finally, a formal system is developed to collect, analyze, and use disciplinary data (e.g., office discipline referrals) to inform decision-making. This presentation provides a brief overview of PBIS and reports findings from a series of four U.S. based longitudinal randomized controlled trials (RCTs) documenting the impacts of PBIS on school climate, discipline problems, bullying, and academic achievement. The four RCTs include 80 elementary, 40 middle, and 58 high schools and results indicate a broad range of impacts on multiple student and school-wide outcomes. The session will highlight lessons learned regarding PBIS implementation and scale-up. We also review the ways in which PBIS can help educators and school leaders engage in data-based decision-making and share data with other decision-makers and stakeholders (e.g., students, parents, community members), with the overarching goal of increasing use of evidence-based programs in schools.

Keywords: positive behavioral interventions and supports, mental health, randomized trials, school-based prevention

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22 Spectroscopic Study of the Anti-Inflammatory Action of Propofol and Its Oxidant Derivatives: Inhibition of the Myeloperoxidase Activity and of the Superoxide Anions Production by Neutrophils

Authors: Pauline Nyssen, Ange Mouithys-Mickalad, Maryse Hoebeke

Abstract:

Inflammation is a complex physiological phenomenon involving chemical and enzymatic mechanisms. Polymorphonuclear neutrophil leukocytes (PMNs) play an important role by producing reactive oxygen species (ROS) and releasing myeloperoxidase (MPO), a pro-oxidant enzyme. Released both in the phagolysosome and the extracellular medium, MPO produces during its peroxidase and halogenation cycles oxidant species, including hypochlorous acid, involved in the destruction of pathogen agents, like bacteria or viruses. Inflammatory pathologies, like rheumatoid arthritis, atherosclerosis induce an excessive stimulation of the PMNs and, therefore, an uncontrolled release of ROS and MPO in the extracellular medium, causing severe damages to the surrounding tissues and biomolecules such as proteins, lipids, and DNA. The treatment of chronic inflammatory pathologies remains a challenge. For many years, MPO has been used as a target for the development of effective treatments. Numerous studies have been focused on the design of new drugs presenting more efficient MPO inhibitory properties. However, some designed inhibitors can be toxic. An alternative consists of assessing the potential inhibitory action of clinically-known molecules, having antioxidant activity. Propofol, 2,6-diisopropyl phenol, which is used as an intravenous anesthetic agent, meets these requirements. Besides its anesthetic action employed to induce a sedative state during surgery or in intensive care units, propofol and its injectable form Diprivan indeed present antioxidant properties and act as ROS and free radical scavengers. A study has also evidenced the ability of propofol to inhibit the formation of the neutrophil extracellular traps fibers, which are important to trap pathogen microorganisms during the inflammation process. The aim of this study was to investigate the potential inhibitory action mechanism of propofol and Diprivan on MPO activity. To go into the anti-inflammatory action of propofol in-depth, two of its oxidative derivatives, 2,6-diisopropyl-1,4-p-benzoquinone (PPFQ) and 3,5,3’,5’-tetra isopropyl-(4,4’)-diphenoquinone (PPFDQ), were studied regarding their inhibitory action. Specific immunological extraction followed by enzyme detection (SIEFED) and molecular modeling have evidenced the low anti-catalytic action of propofol. Stopped-flow absorption spectroscopy and direct MPO activity analysis have proved that propofol acts as a reversible MPO inhibitor by interacting as a reductive substrate in the peroxidase cycle and promoting the accumulation of redox compound II. Overall, Diprivan exhibited a weaker inhibitory action than the active molecule propofol. In contrast, PPFQ seemed to bind and obstruct the enzyme active site, preventing the trigger of the MPO oxidant cycles. PPFQ induced a better chlorination cycle inhibition at basic and neutral pH in comparison to propofol. PPFDQ did not show any MPO inhibition activity. The three interest molecules have also demonstrated their inhibition ability on an important step of the inflammation pathway, the PMNs superoxide anions production, thanks to EPR spectroscopy and chemiluminescence. In conclusion, propofol presents an interesting immunomodulatory activity by acting as a reductive substrate in the peroxidase cycle of MPO, slowing down its activity, whereas PPFQ acts more as an anti-catalytic substrate. Although PPFDQ has no impact on MPO, it can act on the inflammation process by inhibiting the superoxide anions production by PMNs.

Keywords: Diprivan, inhibitor, myeloperoxidase, propofol, spectroscopy

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21 Image Segmentation with Deep Learning of Prostate Cancer Bone Metastases on Computed Tomography

Authors: Joseph M. Rich, Vinay A. Duddalwar, Assad A. Oberai

Abstract:

Prostate adenocarcinoma is the most common cancer in males, with osseous metastases as the commonest site of metastatic prostate carcinoma (mPC). Treatment monitoring is based on the evaluation and characterization of lesions on multiple imaging studies, including Computed Tomography (CT). Monitoring of the osseous disease burden, including follow-up of lesions and identification and characterization of new lesions, is a laborious task for radiologists. Deep learning algorithms are increasingly used to perform tasks such as identification and segmentation for osseous metastatic disease and provide accurate information regarding metastatic burden. Here, nnUNet was used to produce a model which can segment CT scan images of prostate adenocarcinoma vertebral bone metastatic lesions. nnUNet is an open-source Python package that adds optimizations to deep learning-based UNet architecture but has not been extensively combined with transfer learning techniques due to the absence of a readily available functionality of this method. The IRB-approved study data set includes imaging studies from patients with mPC who were enrolled in clinical trials at the University of Southern California (USC) Health Science Campus and Los Angeles County (LAC)/USC medical center. Manual segmentation of metastatic lesions was completed by an expert radiologist Dr. Vinay Duddalwar (20+ years in radiology and oncologic imaging), to serve as ground truths for the automated segmentation. Despite nnUNet’s success on some medical segmentation tasks, it only produced an average Dice Similarity Coefficient (DSC) of 0.31 on the USC dataset. DSC results fell in a bimodal distribution, with most scores falling either over 0.66 (reasonably accurate) or at 0 (no lesion detected). Applying more aggressive data augmentation techniques dropped the DSC to 0.15, and reducing the number of epochs reduced the DSC to below 0.1. Datasets have been identified for transfer learning, which involve balancing between size and similarity of the dataset. Identified datasets include the Pancreas data from the Medical Segmentation Decathlon, Pelvic Reference Data, and CT volumes with multiple organ segmentations (CT-ORG). Some of the challenges of producing an accurate model from the USC dataset include small dataset size (115 images), 2D data (as nnUNet generally performs better on 3D data), and the limited amount of public data capturing annotated CT images of bone lesions. Optimizations and improvements will be made by applying transfer learning and generative methods, including incorporating generative adversarial networks and diffusion models in order to augment the dataset. Performance with different libraries, including MONAI and custom architectures with Pytorch, will be compared. In the future, molecular correlations will be tracked with radiologic features for the purpose of multimodal composite biomarker identification. Once validated, these models will be incorporated into evaluation workflows to optimize radiologist evaluation. Our work demonstrates the challenges of applying automated image segmentation to small medical datasets and lays a foundation for techniques to improve performance. As machine learning models become increasingly incorporated into the workflow of radiologists, these findings will help improve the speed and accuracy of vertebral metastatic lesions detection.

Keywords: deep learning, image segmentation, medicine, nnUNet, prostate carcinoma, radiomics

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20 The Routes of Human Suffering: How Point-Source and Destination-Source Mapping Can Help Victim Services Providers and Law Enforcement Agencies Effectively Combat Human Trafficking

Authors: Benjamin Thomas Greer, Grace Cotulla, Mandy Johnson

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Human trafficking is one of the fastest growing international crimes and human rights violations in the world. The United States Department of State (State Department) approximates some 800,000 to 900,000 people are annually trafficked across sovereign borders, with approximately 14,000 to 17,500 of these people coming into the United States. Today’s slavery is conducted by unscrupulous individuals who are often connected to organized criminal enterprises and transnational gangs, extracting huge monetary sums. According to the International Labour Organization (ILO), human traffickers collect approximately $32 billion worldwide annually. Surpassed only by narcotics dealing, trafficking of humans is tied with illegal arms sales as the second largest criminal industry in the world and is the fastest growing field in the 21st century. Perpetrators of this heinous crime abound. They are not limited to single or “sole practitioners” of human trafficking, but rather, often include Transnational Criminal Organizations (TCO), domestic street gangs, labor contractors, and otherwise seemingly ordinary citizens. Monetary gain is being elevated over territorial disputes and street gangs are increasingly operating in a collaborative effort with TCOs to further disguise their criminal activity; to utilizing their vast networks, in an attempt to avoid detection. Traffickers rely on a network of clandestine routes to sell their commodities with impunity. As law enforcement agencies seek to retard the expansion of transnational criminal organization’s entry into human trafficking, it is imperative that they develop reliable trafficking mapping of known exploitative routes. In a recent report given to the Mexican Congress, The Procuraduría General de la República (PGR) disclosed, from 2008 to 2010 they had identified at least 47 unique criminal networking routes used to traffic victims and that Mexico’s estimated domestic victims number between 800,000 adults and 20,000 children annually. Designing a reliable mapping system is a crucial step to effective law enforcement response and deploying a successful victim support system. Creating this mapping analytic is exceedingly difficult. Traffickers are constantly changing the way they traffic and exploit their victims. They swiftly adapt to local environmental factors and react remarkably well to market demands, exploiting limitations in the prevailing laws. This article will highlight how human trafficking has become one of the fastest growing and most high profile human rights violations in the world today; compile current efforts to map and illustrate trafficking routes; and will demonstrate how the proprietary analytical mapping analysis of point-source and destination-source mapping can help local law enforcement, governmental agencies and victim services providers effectively respond to the type and nature of trafficking to their specific geographical locale. Trafficking transcends state and international borders. It demands an effective and consistent cooperation between local, state, and federal authorities. Each region of the world has different impact factors which create distinct challenges for law enforcement and victim services. Our mapping system lays the groundwork for a targeted anti-trafficking response.

Keywords: human trafficking, mapping, routes, law enforcement intelligence

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19 Characterizing and Developing the Clinical Grade Microbiome Assay with a Robust Bioinformatics Pipeline for Supporting Precision Medicine Driven Clinical Development

Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

Abstract:

Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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18 Magnetic Single-Walled Carbon Nanotubes (SWCNTs) as Novel Theranostic Nanocarriers: Enhanced Targeting and Noninvasive MRI Tracking

Authors: Achraf Al Faraj, Asma Sultana Shaik, Baraa Al Sayed

Abstract:

Specific and effective targeting of drug delivery systems (DDS) to cancerous sites remains a major challenge for a better diagnostic and therapy. Recently, SWCNTs with their unique physicochemical properties and the ability to cross the cell membrane show promising in the biomedical field. The purpose of this study was first to develop a biocompatible iron oxide tagged SWCNTs as diagnostic nanoprobes to allow their noninvasive detection using MRI and their preferential targeting in a breast cancer murine model by placing an optimized flexible magnet over the tumor site. Magnetic targeting was associated to specific antibody-conjugated SWCNTs active targeting. The therapeutic efficacy of doxorubicin-conjugated SWCNTs was assessed, and the superiority of diffusion-weighted (DW-) MRI as sensitive imaging biomarker was investigated. Short Polyvinylpyrrolidone (PVP) stabilized water soluble SWCNTs were first developed, tagged with iron oxide nanoparticles and conjugated with Endoglin/CD105 monoclonal antibodies. They were then conjugated with doxorubicin drugs. SWCNTs conjugates were extensively characterized using TEM, UV-Vis spectrophotometer, dynamic light scattering (DLS) zeta potential analysis and electron spin resonance (ESR) spectroscopy. Their MR relaxivities (i.e. r1 and r2*) were measured at 4.7T and their iron content and metal impurities quantified using ICP-MS. SWCNTs biocompatibility and drug efficacy were then evaluated both in vitro and in vivo using a set of immunological assays. Luciferase enhanced bioluminescence 4T1 mouse mammary tumor cells (4T1-Luc2) were injected into the right inguinal mammary fat pad of Balb/c mice. Tumor bearing mice received either free doxorubicin (DOX) drug or SWCNTs with or without either DOX or iron oxide nanoparticles. A multi-pole 10x10mm high-energy flexible magnet was maintained over the tumor site during 2 hours post-injections and their properties and polarity were optimized to allow enhanced magnetic targeting of SWCNTs toward the primary tumor site. Tumor volume was quantified during the follow-up investigation study using a fast spin echo MRI sequence. In order to detect the homing of SWCNTs to the main tumor site, susceptibility-weighted multi-gradient echo (MGE) sequence was used to generate T2* maps. Apparent diffusion coefficient (ADC) measurements were also performed as a sensitive imaging biomarker providing early and better assessment of disease treatment. At several times post-SWCNT injection, histological analysis were performed on tumor extracts and iron-loaded SWCNT were quantified using ICP-MS in tumor sites, liver, spleen, kidneys, and lung. The optimized multi-poles magnet revealed an enhanced targeting of magnetic SWCNTs to the primary tumor site, which was found to be much higher than the active targeting achieved using antibody-conjugated SWCNTs. Iron-loading allowed their sensitive noninvasive tracking after intravenous administration using MRI. The active targeting of doxorubicin through magnetic antibody-conjugated SWCNTs nanoprobes was found to considerably decrease the primary tumor site and may have inhibited the development of metastasis in the tumor-bearing mice lung. ADC measurements in DW-MRI were found to significantly increase in a time-dependent manner after the injection of DOX-conjugated SWCNTs complexes.

Keywords: single-walled carbon nanotubes, nanomedicine, magnetic resonance imaging, cancer diagnosis and therapy

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17 Analysis Of Fine Motor Skills in Chronic Neurodegenerative Models of Huntington’s Disease and Amyotrophic Lateral Sclerosis

Authors: T. Heikkinen, J. Oksman, T. Bragge, A. Nurmi, O. Kontkanen, T. Ahtoniemi

Abstract:

Motor impairment is an inherent phenotypic feature of several chronic neurodegenerative diseases, and pharmacological therapies aimed to counterbalance the motor disability have a great market potential. Animal models of chronic neurodegenerative diseases display a number deteriorating motor phenotype during the disease progression. There is a wide array of behavioral tools to evaluate motor functions in rodents. However, currently existing methods to study motor functions in rodents are often limited to evaluate gross motor functions only at advanced stages of the disease phenotype. The most commonly applied traditional motor assays used in CNS rodent models, lack the sensitivity to capture fine motor impairments or improvements. Fine motor skill characterization in rodents provides a more sensitive tool to capture more subtle motor dysfunctions and therapeutic effects. Importantly, similar approach, kinematic movement analysis, is also used in clinic, and applied both in diagnosis and determination of therapeutic response to pharmacological interventions. The aim of this study was to apply kinematic gait analysis, a novel and automated high precision movement analysis system, to characterize phenotypic deficits in three different chronic neurodegenerative animal models, a transgenic mouse model (SOD1 G93A) for amyotrophic lateral sclerosis (ALS), and R6/2 and Q175KI mouse models for Huntington’s disease (HD). The readouts from walking behavior included gait properties with kinematic data, and body movement trajectories including analysis of various points of interest such as movement and position of landmarks in the torso, tail and joints. Mice (transgenic and wild-type) from each model were analyzed for the fine motor kinematic properties at young ages, prior to the age when gross motor deficits are clearly pronounced. Fine motor kinematic Evaluation was continued in the same animals until clear motor dysfunction with conventional motor assays was evident. Time course analysis revealed clear fine motor skill impairments in each transgenic model earlier than what is seen with conventional gross motor tests. Motor changes were quantitatively analyzed for up to ~80 parameters, and the largest data sets of HD models were further processed with principal component analysis (PCA) to transform the pool of individual parameters into a smaller and focused set of mutually uncorrelated gait parameters showing strong genotype difference. Kinematic fine motor analysis of transgenic animal models described in this presentation show that this method isa sensitive, objective and fully automated tool that allows earlier and more sensitive detection of progressive neuromuscular and CNS disease phenotypes. As a result of the analysis a comprehensive set of fine motor parameters for each model is created, and these parameters provide better understanding of the disease progression and enhanced sensitivity of this assay for therapeutic testing compared to classical motor behavior tests. In SOD1 G93A, R6/2, and Q175KI mice, the alterations in gait were evident already several weeks earlier than with traditional gross motor assays. Kinematic testing can be applied to a wider set of motor readouts beyond gait in order to study whole body movement patterns such as with relation to joints and various body parts longitudinally, providing a sophisticated and translatable method for disseminating motor components in rodent disease models and evaluating therapeutic interventions.

Keywords: Gait analysis, kinematic, motor impairment, inherent feature

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16 Surface Sunctionalization Strategies for the Design of Thermoplastic Microfluidic Devices for New Analytical Diagnostics

Authors: Camille Perréard, Yoann Ladner, Fanny D'Orlyé, Stéphanie Descroix, Vélan Taniga, Anne Varenne, Cédric Guyon, Michael. Tatoulian, Frédéric Kanoufi, Cyrine Slim, Sophie Griveau, Fethi Bedioui

Abstract:

The development of micro total analysis systems is of major interest for contaminant and biomarker analysis. As a lab-on-chip integrates all steps of an analysis procedure in a single device, analysis can be performed in an automated format with reduced time and cost, while maintaining performances comparable to those of conventional chromatographic systems. Moreover, these miniaturized systems are either compatible with field work or glovebox manipulations. This work is aimed at developing an analytical microsystem for trace and ultra trace quantitation in complex matrices. The strategy consists in the integration of a sample pretreatment step within the lab-on-chip by a confinement zone where selective ligands are immobilized for target extraction and preconcentration. Aptamers were chosen as selective ligands, because of their high affinity for all types of targets (from small ions to viruses and cells) and their ease of synthesis and functionalization. This integrated target extraction and concentration step will be followed in the microdevice by an electrokinetic separation step and an on-line detection. Polymers consisting of cyclic olefin copolymer (COC) or fluoropolymer (Dyneon THV) were selected as they are easy to mold, transparent in UV-visible and have high resistance towards solvents and extreme pH conditions. However, because of their low chemical reactivity, surface treatments are necessary. For the design of this miniaturized diagnostics, we aimed at modifying the microfluidic system at two scales : (1) on the entire surface of the microsystem to control the surface hydrophobicity (so as to avoid any sample wall adsorption) and the fluid flows during electrokinetic separation, or (2) locally so as to immobilize selective ligands (aptamers) on restricted areas for target extraction and preconcentration. We developed different novel strategies for the surface functionalization of COC and Dyneon, based on plasma, chemical and /or electrochemical approaches. In a first approach, a plasma-induced immobilization of brominated derivatives was performed on the entire surface. Further substitution of the bromine by an azide functional group led to covalent immobilization of ligands through “click” chemistry reaction between azides and terminal alkynes. COC and Dyneon materials were characterized at each step of the surface functionalization procedure by various complementary techniques to evaluate the quality and homogeneity of the functionalization (contact angle, XPS, ATR). With the objective of local (micrometric scale) aptamer immobilization, we developed an original electrochemical strategy on engraved Dyneon THV microchannel. Through local electrochemical carbonization followed by adsorption of azide-bearing diazonium moieties and covalent linkage of alkyne-bearing aptamers through click chemistry reaction, typical dimensions of immobilization zones reached the 50 µm range. Other functionalization strategies, such as sol-gel encapsulation of aptamers, are currently investigated and may also be suitable for the development of the analytical microdevice. The development of these functionalization strategies is the first crucial step in the design of the entire microdevice. These strategies allow the grafting of a large number of molecules for the development of new analytical tools in various domains like environment or healthcare.

Keywords: alkyne-azide click chemistry (CuAAC), electrochemical modification, microsystem, plasma bromination, surface functionalization, thermoplastic polymers

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15 Smart Interior Design: A Revolution in Modern Living

Authors: Fatemeh Modirzare

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Smart interior design represents a transformative approach to creating living spaces that integrate technology seamlessly into our daily lives, enhancing comfort, convenience, and sustainability. This paper explores the concept of smart interior design, its principles, benefits, challenges, and future prospects. It also highlights various examples and applications of smart interior design to illustrate its potential in shaping the way we live and interact with our surroundings. In an increasingly digitized world, the boundaries between technology and interior design are blurring. Smart interior design, also known as intelligent or connected interior design, involves the incorporation of advanced technologies and automation systems into residential and commercial spaces. This innovative approach aims to make living environments more efficient, comfortable, and adaptable while promoting sustainability and user well-being. Smart interior design seamlessly integrates technology into the aesthetics and functionality of a space, ensuring that devices and systems do not disrupt the overall design. Sustainable materials, energy-efficient systems, and eco-friendly practices are central to smart interior design, reducing environmental impact. Spaces are designed to be adaptable, allowing for reconfiguration to suit changing needs and preferences. Smart homes and spaces offer greater comfort through features like automated climate control, adjustable lighting, and customizable ambiance. Smart interior design can significantly reduce energy consumption through optimized heating, cooling, and lighting systems. Smart interior design integrates security systems, fire detection, and emergency response mechanisms for enhanced safety. Sustainable materials, energy-efficient appliances, and waste reduction practices contribute to a greener living environment. Implementing smart interior design can be expensive, particularly when retrofitting existing spaces with smart technologies. The increased connectivity raises concerns about data privacy and cybersecurity, requiring robust measures to protect user information. Rapid advancements in technology may lead to obsolescence, necessitating updates and replacements. Users must be familiar with smart systems to fully benefit from them, requiring education and ongoing support. Residential spaces incorporate features like voice-activated assistants, automated lighting, and energy management systems. Intelligent office design enhances productivity and employee well-being through smart lighting, climate control, and meeting room booking systems. Hospitals and healthcare facilities use smart interior design for patient monitoring, wayfinding, and energy conservation. Smart retail design includes interactive displays, personalized shopping experiences, and inventory management systems. The future of smart interior design holds exciting possibilities, including AI-powered design tools that create personalized spaces based on user preferences. Smart interior design will increasingly prioritize factors that improve physical and mental health, such as air quality monitoring and mood-enhancing lighting. Smart interior design is revolutionizing the way we interact with our living and working spaces. By embracing technology, sustainability, and user-centric design principles, smart interior design offers numerous benefits, from increased comfort and convenience to energy efficiency and sustainability. Despite challenges, the future holds tremendous potential for further innovation in this field, promising a more connected, efficient, and harmonious way of living and working.

Keywords: smart interior design, home automation, sustainable living spaces, technological integration, user-centric design

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14 Sinhala Sign Language to Grammatically Correct Sentences using NLP

Authors: Anjalika Fernando, Banuka Athuraliya

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This paper presents a comprehensive approach for converting Sinhala Sign Language (SSL) into grammatically correct sentences using Natural Language Processing (NLP) techniques in real-time. While previous studies have explored various aspects of SSL translation, the research gap lies in the absence of grammar checking for SSL. This work aims to bridge this gap by proposing a two-stage methodology that leverages deep learning models to detect signs and translate them into coherent sentences, ensuring grammatical accuracy. The first stage of the approach involves the utilization of a Long Short-Term Memory (LSTM) deep learning model to recognize and interpret SSL signs. By training the LSTM model on a dataset of SSL gestures, it learns to accurately classify and translate these signs into textual representations. The LSTM model achieves a commendable accuracy rate of 94%, demonstrating its effectiveness in accurately recognizing and translating SSL gestures. Building upon the successful recognition and translation of SSL signs, the second stage of the methodology focuses on improving the grammatical correctness of the translated sentences. The project employs a Neural Machine Translation (NMT) architecture, consisting of an encoder and decoder with LSTM components, to enhance the syntactical structure of the generated sentences. By training the NMT model on a parallel corpus of Sinhala wrong sentences and their corresponding grammatically correct translations, it learns to generate coherent and grammatically accurate sentences. The NMT model achieves an impressive accuracy rate of 98%, affirming its capability to produce linguistically sound translations. The proposed approach offers significant contributions to the field of SSL translation and grammar correction. Addressing the critical issue of grammar checking, it enhances the usability and reliability of SSL translation systems, facilitating effective communication between hearing-impaired and non-sign language users. Furthermore, the integration of deep learning techniques, such as LSTM and NMT, ensures the accuracy and robustness of the translation process. This research holds great potential for practical applications, including educational platforms, accessibility tools, and communication aids for the hearing-impaired. Furthermore, it lays the foundation for future advancements in SSL translation systems, fostering inclusive and equal opportunities for the deaf community. Future work includes expanding the existing datasets to further improve the accuracy and generalization of the SSL translation system. Additionally, the development of a dedicated mobile application would enhance the accessibility and convenience of SSL translation on handheld devices. Furthermore, efforts will be made to enhance the current application for educational purposes, enabling individuals to learn and practice SSL more effectively. Another area of future exploration involves enabling two-way communication, allowing seamless interaction between sign-language users and non-sign-language users.In conclusion, this paper presents a novel approach for converting Sinhala Sign Language gestures into grammatically correct sentences using NLP techniques in real time. The two-stage methodology, comprising an LSTM model for sign detection and translation and an NMT model for grammar correction, achieves high accuracy rates of 94% and 98%, respectively. By addressing the lack of grammar checking in existing SSL translation research, this work contributes significantly to the development of more accurate and reliable SSL translation systems, thereby fostering effective communication and inclusivity for the hearing-impaired community

Keywords: Sinhala sign language, sign Language, NLP, LSTM, NMT

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13 Blue Economy and Marine Mining

Authors: Fani Sakellariadou

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The Blue Economy includes all marine-based and marine-related activities. They correspond to established, emerging as well as unborn ocean-based industries. Seabed mining is an emerging marine-based activity; its operations depend particularly on cutting-edge science and technology. The 21st century will face a crisis in resources as a consequence of the world’s population growth and the rising standard of living. The natural capital stored in the global ocean is decisive for it to provide a wide range of sustainable ecosystem services. Seabed mineral deposits were identified as having a high potential for critical elements and base metals. They have a crucial role in the fast evolution of green technologies. The major categories of marine mineral deposits are deep-sea deposits, including cobalt-rich ferromanganese crusts, polymetallic nodules, phosphorites, and deep-sea muds, as well as shallow-water deposits including marine placers. Seabed mining operations may take place within continental shelf areas of nation-states. In international waters, the International Seabed Authority (ISA) has entered into 15-year contracts for deep-seabed exploration with 21 contractors. These contracts are for polymetallic nodules (18 contracts), polymetallic sulfides (7 contracts), and cobalt-rich ferromanganese crusts (5 contracts). Exploration areas are located in the Clarion-Clipperton Zone, the Indian Ocean, the Mid Atlantic Ridge, the South Atlantic Ocean, and the Pacific Ocean. Potential environmental impacts of deep-sea mining include habitat alteration, sediment disturbance, plume discharge, toxic compounds release, light and noise generation, and air emissions. They could cause burial and smothering of benthic species, health problems for marine species, biodiversity loss, reduced photosynthetic mechanism, behavior change and masking acoustic communication for mammals and fish, heavy metals bioaccumulation up the food web, decrease of the content of dissolved oxygen, and climate change. An important concern related to deep-sea mining is our knowledge gap regarding deep-sea bio-communities. The ecological consequences that will be caused in the remote, unique, fragile, and little-understood deep-sea ecosystems and inhabitants are still largely unknown. The blue economy conceptualizes oceans as developing spaces supplying socio-economic benefits for current and future generations but also protecting, supporting, and restoring biodiversity and ecological productivity. In that sense, people should apply holistic management and make an assessment of marine mining impacts on ecosystem services, including the categories of provisioning, regulating, supporting, and cultural services. The variety in environmental parameters, the range in sea depth, the diversity in the characteristics of marine species, and the possible proximity to other existing maritime industries cause a span of marine mining impact the ability of ecosystems to support people and nature. In conclusion, the use of the untapped potential of the global ocean demands a liable and sustainable attitude. Moreover, there is a need to change our lifestyle and move beyond the philosophy of single-use. Living in a throw-away society based on a linear approach to resource consumption, humans are putting too much pressure on the natural environment. Applying modern, sustainable and eco-friendly approaches according to the principle of circular economy, a substantial amount of natural resource savings will be achieved. Acknowledgement: This work is part of the MAREE project, financially supported by the Division VI of IUPAC. This work has been partly supported by the University of Piraeus Research Center.

Keywords: blue economy, deep-sea mining, ecosystem services, environmental impacts

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12 A Human Factors Approach to Workload Optimization for On-Screen Review Tasks

Authors: Christina Kirsch, Adam Hatzigiannis

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Rail operators and maintainers worldwide are increasingly replacing walking patrols in the rail corridor with mechanized track patrols -essentially data capture on trains- and on-screen reviews of track infrastructure in centralized review facilities. The benefit is that infrastructure workers are less exposed to the dangers of the rail corridor. The impact is a significant change in work design from walking track sections and direct observation in the real world to sedentary jobs in the review facility reviewing captured data on screens. Defects in rail infrastructure can have catastrophic consequences. Reviewer performance regarding accuracy and efficiency of reviews within the available time frame is essential to ensure safety and operational performance. Rail operators must optimize workload and resource loading to transition to on-screen reviews successfully. Therefore, they need to know what workload assessment methodologies will provide reliable and valid data to optimize resourcing for on-screen reviews. This paper compares objective workload measures, including track difficulty ratings and review distance covered per hour, and subjective workload assessments (NASA TLX) and analyses the link between workload and reviewer performance, including sensitivity, precision, and overall accuracy. An experimental study was completed with eight on-screen reviewers, including infrastructure workers and engineers, reviewing track sections with different levels of track difficulty over nine days. Each day the reviewers completed four 90-minute sessions of on-screen inspection of the track infrastructure. Data regarding the speed of review (km/ hour), detected defects, false negatives, and false positives were collected. Additionally, all reviewers completed a subjective workload assessment (NASA TLX) after each 90-minute session and a short employee engagement survey at the end of the study period that captured impacts on job satisfaction and motivation. The results showed that objective measures for tracking difficulty align with subjective mental demand, temporal demand, effort, and frustration in the NASA TLX. Interestingly, review speed correlated with subjective assessments of physical and temporal demand, but to mental demand. Subjective performance ratings correlated with all accuracy measures and review speed. The results showed that subjective NASA TLX workload assessments accurately reflect objective workload. The analysis of the impact of workload on performance showed that subjective mental demand correlated with high precision -accurately detected defects, not false positives. Conversely, high temporal demand was negatively correlated with sensitivity and the percentage of detected existing defects. Review speed was significantly correlated with false negatives. With an increase in review speed, accuracy declined. On the other hand, review speed correlated with subjective performance assessments. Reviewers thought their performance was higher when they reviewed the track sections faster, despite the decline in accuracy. The study results were used to optimize resourcing and ensure that reviewers had enough time to review the allocated track sections to improve defect detection rates in accordance with the efficiency-thoroughness trade-off. Overall, the study showed the importance of a multi-method approach to workload assessment and optimization, combining subjective workload assessments with objective workload and performance measures to ensure that recommendations for work system optimization are evidence-based and reliable.

Keywords: automation, efficiency-thoroughness trade-off, human factors, job design, NASA TLX, performance optimization, subjective workload assessment, workload analysis

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11 Settings of Conditions Leading to Reproducible and Robust Biofilm Formation in vitro in Evaluation of Drug Activity against Staphylococcal Biofilms

Authors: Adela Diepoltova, Klara Konecna, Ondrej Jandourek, Petr Nachtigal

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A loss of control over antibiotic-resistant pathogens has become a global issue due to severe and often untreatable infections. This state is reflected in complicated treatment, health costs, and higher mortality. All these factors emphasize the urgent need for the discovery and development of new anti-infectives. One of the most common pathogens mentioned in the phenomenon of antibiotic resistance are bacteria of the genus Staphylococcus. These bacterial agents have developed several mechanisms against the effect of antibiotics. One of them is biofilm formation. In staphylococci, biofilms are associated with infections such as endocarditis, osteomyelitis, catheter-related bloodstream infections, etc. To author's best knowledge, no validated and standardized methodology evaluating candidate compound activity against staphylococcal biofilms exists. However, a variety of protocols for in vitro drug activity testing has been suggested, yet there are often fundamental differences. Based on our experience, a key methodological step that leads to credible results is to form a robust biofilm with appropriate attributes such as firm adherence to the substrate, a complex arrangement in layers, and the presence of extracellular polysaccharide matrix. At first, for the purpose of drug antibiofilm activity evaluation, the focus was put on various conditions (supplementation of cultivation media by human plasma/fetal bovine serum, shaking mode, the density of initial inoculum) that should lead to reproducible and robust in vitro staphylococcal biofilm formation in microtiter plate model. Three model staphylococcal reference strains were included in the study: Staphylococcus aureus (ATCC 29213), methicillin-resistant Staphylococcus aureus (ATCC 43300), and Staphylococcus epidermidis (ATCC 35983). The total biofilm biomass was quantified using the Christensen method with crystal violet, and results obtained from at least three independent experiments were statistically processed. Attention was also paid to the viability of the biofilm-forming staphylococcal cells and the presence of extracellular polysaccharide matrix. The conditions that led to robust biofilm biomass formation with attributes for biofilms mentioned above were then applied by introducing an alternative method analogous to the commercially available test system, the Calgary Biofilm Device. In this test system, biofilms are formed on pegs that are incorporated into the lid of the microtiter plate. This system provides several advantages (in situ detection and quantification of biofilm microbial cells that have retained their viability after drug exposure). Based on our preliminary studies, it was found that the attention to the peg surface and substrate on which the bacterial biofilms are formed should also be paid to. Therefore, further steps leading to the optimization were introduced. The surface of pegs was coated by human plasma, fetal bovine serum, and L-polylysine. Subsequently, the willingness of bacteria to adhere and form biofilm was monitored. In conclusion, suitable conditions were revealed, leading to the formation of reproducible, robust staphylococcal biofilms in vitro for the microtiter model and the system analogous to the Calgary biofilm device, as well. The robustness and typical slime texture could be detected visually. Likewise, an analysis by confocal laser scanning microscopy revealed a complex three-dimensional arrangement of biofilm forming organisms surrounded by an extracellular polysaccharide matrix.

Keywords: anti-biofilm drug activity screening, in vitro biofilm formation, microtiter plate model, the Calgary biofilm device, staphylococcal infections, substrate modification, surface coating

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10 Results concerning the University: Industry Partnership for a Research Project Implementation (MUROS) in the Romanian Program Star

Authors: Loretta Ichim, Dan Popescu, Grigore Stamatescu

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The paper reports the collaboration between a top university from Romania and three companies for the implementation of a research project in a multidisciplinary domain, focusing on the impact and benefits both for the education and industry. The joint activities were developed under the Space Technology and Advanced Research Program (STAR), funded by the Romanian Space Agency (ROSA) for a university-industry partnership. The context was defined by linking the European Space Agency optional programs, with the development and promotion national research, with the educational and industrial capabilities in the aeronautics, security and related areas by increasing the collaboration between academic and industrial entities as well as by realizing high-level scientific production. The project name is Multisensory Robotic System for Aerial Monitoring of Critical Infrastructure Systems (MUROS), which was carried 2013-2016. The project included the University POLITEHNICA of Bucharest (coordinator) and three companies, which manufacture and market unmanned aerial systems. The project had as main objective the development of an integrated system for combined ground wireless sensor networks and UAV monitoring in various application scenarios for critical infrastructure surveillance. This included specific activities related to fundamental and applied research, technology transfer, prototype implementation and result dissemination. The core area of the contributions laid in distributed data processing and communication mechanisms, advanced image processing and embedded system development. Special focus is given by the paper to analyzing the impact the project implementation in the educational process, directly or indirectly, through the faculty members (professors and students) involved in the research team. Three main directions are discussed: a) enabling students to carry out internships at the partner companies, b) handling advanced topics and industry requirements at the master's level, c) experiments and concept validation for doctoral thesis. The impact of the research work (as the educational component) developed by the faculty members on the increasing performances of the companies’ products is highlighted. The collaboration between university and companies was well balanced both for contributions and results. The paper also presents the outcomes of the project which reveals the efficient collaboration between high education and industry: master thesis, doctoral thesis, conference papers, journal papers, technical documentation for technology transfer, prototype, and patent. The experience can provide useful practices of blending research and education within an academia-industry cooperation framework while the lessons learned represent a starting point in debating the new role of advanced research and development performing companies in association with higher education. This partnership, promoted at UE level, has a broad impact beyond the constrained scope of a single project and can develop into long-lasting collaboration while benefiting all stakeholders: students, universities and the surrounding knowledge-based economic and industrial ecosystem. Due to the exchange of experiences between the university (UPB) and the manufacturing company (AFT Design), a new project, SIMUL, under the Bridge Grant Program (Romanian executive agency UEFISCDI) was started (2016 – 2017). This project will continue the educational research for innovation on master and doctoral studies in MUROS thematic (collaborative multi-UAV application for flood detection).

Keywords: education process, multisensory robotic system, research and innovation project, technology transfer, university-industry partnership

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9 Identification Strategies for Unknown Victims from Mass Disasters and Unknown Perpetrators from Violent Crime or Terrorist Attacks

Authors: Michael Josef Schwerer

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Background: The identification of unknown victims from mass disasters, violent crimes, or terrorist attacks is frequently facilitated through information from missing persons lists, portrait photos, old or recent pictures showing unique characteristics of a person such as scars or tattoos, or simply reference samples from blood relatives for DNA analysis. In contrast, the identification or at least the characterization of an unknown perpetrator from criminal or terrorist actions remains challenging, particularly in the absence of material or data for comparison, such as fingerprints, which had been previously stored in criminal records. In scenarios that result in high levels of destruction of the perpetrator’s corpse, for instance, blast or fire events, the chance for a positive identification using standard techniques is further impaired. Objectives: This study shows the forensic genetic procedures in the Legal Medicine Service of the German Air Force for the identification of unknown individuals, including such cases in which reference samples are not available. Scenarios requiring such efforts predominantly involve aircraft crash investigations, which are routinely carried out by the German Air Force Centre of Aerospace Medicine as one of the Institution’s essential missions. Further, casework by military police or military intelligence is supported based on administrative cooperation. In the talk, data from study projects, as well as examples from real casework, will be demonstrated and discussed with the audience. Methods: Forensic genetic identification in our laboratories involves the analysis of Short Tandem Repeats and Single Nucleotide Polymorphisms in nuclear DNA along with mitochondrial DNA haplotyping. Extended DNA analysis involves phenotypic markers for skin, hair, and eye color together with the investigation of a person’s biogeographic ancestry. Assessment of the biological age of an individual employs CpG-island methylation analysis using bisulfite-converted DNA. Forensic Investigative Genealogy assessment allows the detection of an unknown person’s blood relatives in reference databases. Technically, end-point-PCR, real-time PCR, capillary electrophoresis, pyrosequencing as well as next generation sequencing using flow-cell-based and chip-based systems are used. Results and Discussion: Optimization of DNA extraction from various sources, including difficult matrixes like formalin-fixed, paraffin-embedded tissues, degraded specimens from decomposed bodies or from decedents exposed to blast or fire events, provides soil for successful PCR amplification and subsequent genetic profiling. For cases with extremely low yields of extracted DNA, whole genome preamplification protocols are successfully used, particularly regarding genetic phenotyping. Improved primer design for CpG-methylation analysis, together with validated sampling strategies for the analyzed substrates from, e.g., lymphocyte-rich organs, allows successful biological age estimation even in bodies with highly degraded tissue material. Conclusions: Successful identification of unknown individuals or at least their phenotypic characterization using pigmentation markers together with age-informative methylation profiles, possibly supplemented by family tree search employing Forensic Investigative Genealogy, can be provided in specialized laboratories. However, standard laboratory procedures must be adapted to work with difficult and highly degraded sample materials.

Keywords: identification, forensic genetics, phenotypic markers, CPG methylation, biological age estimation, forensic investigative genealogy

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8 Advancing Dialysis Care Access And Health Information Management: A Blueprint For Nairobi Hospital

Authors: Kimberly Winnie Achieng Otieno

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The Nairobi Hospital plays a pivotal role in healthcare provision in East and Central Africa, yet it faces challenges in providing accessible dialysis care. This paper explores strategic interventions to enhance dialysis care, improve access and streamline health information management, with an aim of fostering an integrated and patient-centered healthcare system in our region. Challenges at The Nairobi Hospital The Nairobi Hospital currently grapples with insufficient dialysis machines which results in extended turn around times. This issue stems from both staffing bottle necks and infrastructural limitations given our growing demand for renal care services. Our Paper-based record keeping system and fragmented flow of information downstream hinders the hospital’s ability to manage health data effectively. There is also a need for investment in expanding The Nairobi Hospital dialysis facilities to far reaching communities. Setting up satellite clinics that are closer to people who live in areas far from the main hospital will ensure better access to underserved areas. Community Outreach and Education Implementing education programs on kidney health within local communities is vital for early detection and prevention. Collaborating with local leaders and organizations can establish a proactive approach to renal health hence reducing the demand for acute dialysis interventions. We can amplify this effort by expanding The Nairobi Hospital’s corporate social responsibility outreach program with weekend engagement activities such as walks, awareness classes and fund drives. Enhancing Efficiency in Dialysis Care Demand for dialysis services continues to rise due to an aging Kenyan population and the increasing prevalence of chronic kidney disease (CKD). Present at this years International Nursing Conference are a diverse group of caregivers from around the world who can share with us their process optimization strategies, patient engagement techniques and resource utilization efficiencies to catapult The Nairobi Hospital to the 21st century and beyond. Plans are underway to offer ongoing education opportunities to keep staff updated on best practices and emerging technologies in addition to utilizing a patient feedback mechanisms to identify areas for improvement and enhance satisfaction. Staff empowerment and suggestion boxes address The Nairobi Hospital’s organizational challenges. Current financial constraints may limit a leapfrog in technology integration such as the acquisition of new dialysis machines and an investment in predictive analytics to forecast patient needs and optimize resource allocation. Streamlining Health Information Management Fully embracing a shift to 100% Electronic Health Records (EHRs) is a transformative step toward efficient health information management. Shared information promotes a holistic understanding of patients’ medical history, minimizing redundancies and enhancing overall care quality. To manage the transition to community-based care and EHRs effectively, a phased implementation approach is recommended. Conclusion By strategically enhancing dialysis care access and streamlining health information management, The Nairobi Hospital can strengthen its position as a leading healthcare institution in both East and Central Africa. This comprehensive approach aligns with the hospital’s commitment to providing high-quality, accessible, and patient-centered care in an evolving landscape of healthcare delivery.

Keywords: Africa, urology, diaylsis, healthcare

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