Search results for: X-ray induced photodynamic therapy
Commenced in January 2007
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Paper Count: 4547

Search results for: X-ray induced photodynamic therapy

77 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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76 Transcriptional Differences in B cell Subpopulations over the Course of Preclinical Autoimmunity Development

Authors: Aleksandra Bylinska, Samantha Slight-Webb, Kevin Thomas, Miles Smith, Susan Macwana, Nicolas Dominguez, Eliza Chakravarty, Joan T. Merrill, Judith A. James, Joel M. Guthridge

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Background: Systemic Lupus Erythematosus (SLE) is an interferon-related autoimmune disease characterized by B cell dysfunction. One of the main hallmarks is a loss of tolerance to self-antigens leading to increased levels of autoantibodies against nuclear components (ANAs). However, up to 20% of healthy ANA+ individuals will not develop clinical illness. SLE is more prevalent among women and minority populations (African, Asian American and Hispanics). Moreover, African Americans have a stronger interferon (IFN) signature and develop more severe symptoms. The exact mechanisms involved in ethnicity-dependent B cell dysregulation and the progression of autoimmune disease from ANA+ healthy individuals to clinical disease remains unclear. Methods: Peripheral blood mononuclear cells (PBMCs) from African (AA) and European American (EA) ANA- (n=12), ANA+ (n=12) and SLE (n=12) individuals were assessed by multimodal scRNA-Seq/CITE-Seq methods to examine differential gene signatures in specific B cell subsets. Library preparation was done with a 10X Genomics Chromium according to established protocols and sequenced on Illumina NextSeq. The data were further analyzed for distinct cluster identification and differential gene signatures in the Seurat package in R and pathways analysis was performed using Ingenuity Pathways Analysis (IPA). Results: Comparing all subjects, 14 distinct B cell clusters were identified using a community detection algorithm and visualized with Uniform Manifold Approximation Projection (UMAP). The proportion of each of those clusters varied by disease status and ethnicity. Transitional B cells trended higher in ANA+ healthy individuals, especially in AA. Ribonucleoprotein high population (HNRNPH1 elevated, heterogeneous nuclear ribonucleoprotein, RNP-Hi) of proliferating Naïve B cells were more prevalent in SLE patients, specifically in EA. Interferon-induced protein high population (IFIT-Hi) of Naive B cells are increased in EA ANA- individuals. The proportion of memory B cells and plasma cells clusters tend to be expanded in SLE patients. As anticipated, we observed a higher signature of cytokine-related pathways, especially interferon, in SLE individuals. Pathway analysis among AA individuals revealed an NRF2-mediated Oxidative Stress response signature in the transitional B cell cluster, not seen in EA individuals. TNFR1/2 and Sirtuin Signaling pathway genes were higher in AA IFIT-Hi Naive B cells, whereas they were not detected in EA individuals. Interferon signaling was observed in B cells in both ethnicities. Oxidative phosphorylation was found in age-related B cells (ABCs) for both ethnicities, whereas Death Receptor Signaling was found only in EA patients in these cells. Interferon-related transcription factors were elevated in ABCs and IFIT-Hi Naive B cells in SLE subjects of both ethnicities. Conclusions: ANA+ healthy individuals have altered gene expression pathways in B cells that might drive apoptosis and subsequent clinical autoimmune pathogenesis. Increases in certain regulatory pathways may delay progression to SLE. Further, AA individuals have more elevated activation pathways that may make them more susceptible to SLE.

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75 Developing a Methodology to Examine Psychophysiological Responses during Stress Exposure and Relaxation: An Experimental Paradigm

Authors: M. Velana, G. Rinkenauer

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Nowadays, nurses are facing unprecedented amounts of pressure due to the ongoing global health demands. Work-related stress can cause a high physical and psychological workload, which can lead, in turn, to burnout. On the physiological level, stress triggers an initial activation of the sympathetic nervous and adrenomedullary systems resulting in increases in cardiac activity. Furthermore, activation of the hypothalamus-pituitary-adrenal axis provokes endocrine and immune changes leading to the release of cortisol and cytokines in an effort to re-establish body balance. Based on the current state of the literature, it has been identified that resilience and mindfulness exercises among nurses can effectively decrease stress and improve mood. However, it is still unknown what relaxation techniques would be suitable for and to what extent would be effective to decrease psychophysiological arousal deriving from either a physiological or a psychological stressor. Moreover, although cardiac activity and cortisol are promising candidates to examine the effectiveness of relaxation to reduce stress, it still remains to shed light on the role of cytokines in this process so as to thoroughly understand the body’s response to stress and to relaxation. Therefore, the main aim of the present study is to develop a comprehensive experimental paradigm and assess different relaxation techniques, namely progressive muscle relaxation and a mindfulness exercise originating from cognitive therapy by means of biofeedback, under highly controlled laboratory conditions. An experimental between-subject design will be employed, where 120 participants will be randomized either to a physiological or a psychological stress-related experiment. Particularly, the cold pressor test refers to a procedure in which the participants have to immerse their non-dominant hands into ice water (2-3 °C) for 3 min. The participants are requested to keep their hands in the water throughout the whole duration. However, they can immediately terminate the test in case it would be barely tolerable. A pre-test anticipation phase and a post-stress period of 3 min, respectively, are planned. The Trier Social Stress Test will be employed to induce psychological stress. During this laboratory stressor, the participants are instructed to give a 5-min speech in front of a committee of communication specialists. Before the main task, there is a 10-min anticipation period. Subsequently, participants are requested to perform an unexpected arithmetic task. After stress exposure, the participants will perform one of the relaxation exercises (treatment condition) or watch a neutral video (control condition). Electrocardiography, salivary samples, and self-report will be collected at different time points. The preliminary results deriving from the pilot study showed that the aforementioned paradigm could effectively induce stress reactions and that relaxation might decrease the impact of stress exposure. It is of utmost importance to assess how the human body responds under different stressors and relaxation exercises so that an evidence-based intervention could be transferred in a clinical setting to improve nurses’ general health. Based on suggestive future laboratory findings, the research group plans to conduct a pilot-level randomized study to decrease stress and promote well-being among nurses who work in the stress-riddled environment of a hospital located in Northern Germany.

Keywords: nurses, psychophysiology, relaxation, stress

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74 Broad Host Range Bacteriophage Cocktail for Reduction of Staphylococcus aureus as Potential Therapy for Atopic Dermatitis

Authors: Tamar Lin, Nufar Buchshtab, Yifat Elharar, Julian Nicenboim, Rotem Edgar, Iddo Weiner, Lior Zelcbuch, Ariel Cohen, Sharon Kredo-Russo, Inbar Gahali-Sass, Naomi Zak, Sailaja Puttagunta, Merav Bassan

Abstract:

Background: Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disorder that is characterized by dry skin and flares of eczematous lesions and intense pruritus. Multiple lines of evidence suggest that AD is associated with increased colonization by Staphylococcus aureus, which contributes to disease pathogenesis through the release of virulence factors that affect both keratinocytes and immune cells, leading to disruption of the skin barrier and immune cell dysfunction. The aim of the current study is to develop a bacteriophage-based product that specifically targets S. aureus. Methods: For the discovery of phage, environmental samples were screened on 118 S. aureus strains isolated from skin samples, followed by multiple enrichment steps. Natural phages were isolated, subjected to Next-generation Sequencing (NGS), and analyzed using proprietary bioinformatics tools for undesirable genes (toxins, antibiotic resistance genes, lysogeny potential), taxonomic classification, and purity. Phage host range was determined by an efficiency of plating (EOP) value above 0.1 and the ability of the cocktail to completely lyse liquid bacterial culture under different growth conditions (e.g., temperature, bacterial stage). Results: Sequencing analysis demonstrated that the 118 S. aureus clinical strains were distributed across the phylogenetic tree of all available Refseq S. aureus (~10,750 strains). Screening environmental samples on the S. aureus isolates resulted in the isolation of 50 lytic phages from different genera, including Silviavirus, Kayvirus, Podoviridae, and a novel unidentified phage. NGS sequencing confirmed the absence of toxic elements in the phages’ genomes. The host range of the individual phages, as measured by the efficiency of plating (EOP), ranged between 41% (48/118) to 79% (93/118). Host range studies in liquid culture revealed that a subset of the phages can infect a broad range of S. aureus strains in different metabolic states, including stationary state. Combining the single-phage EOP results of selected phages resulted in a broad host range cocktail which infected 92% (109/118) of the strains. When tested in vitro in a liquid infection assay, clearance was achieved in 87% (103/118) of the strains, with no evidence of phage resistance throughout the study (24 hours). A S. aureus host was identified that can be used for the production of all the phages in the cocktail at high titers suitable for large-scale manufacturing. This host was validated for the absence of contaminating prophages using advanced NGS methods combined with multiple production cycles. The phages are produced under optimized scale-up conditions and are being used for the development of a topical formulation (BX005) that may be administered to subjects with atopic dermatitis. Conclusions: A cocktail of natural phages targeting S. aureus was effective in reducing bacterial burden across multiple assays. Phage products may offer safe and effective steroid-sparing options for atopic dermatitis.

Keywords: atopic dermatitis, bacteriophage cocktail, host range, Staphylococcus aureus

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73 Optical Imaging Based Detection of Solder Paste in Printed Circuit Board Jet-Printing Inspection

Authors: D. Heinemann, S. Schramm, S. Knabner, D. Baumgarten

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Purpose: Applying solder paste to printed circuit boards (PCB) with stencils has been the method of choice over the past years. A new method uses a jet printer to deposit tiny droplets of solder paste through an ejector mechanism onto the board. This allows for more flexible PCB layouts with smaller components. Due to the viscosity of the solder paste, air blisters can be trapped in the cartridge. This can lead to missing solder joints or deviations in the applied solder volume. Therefore, a built-in and real-time inspection of the printing process is needed to minimize uncertainties and increase the efficiency of the process by immediate correction. The objective of the current study is the design of an optimal imaging system and the development of an automatic algorithm for the detection of applied solder joints from optical from the captured images. Methods: In a first approach, a camera module connected to a microcomputer and LED strips are employed to capture images of the printed circuit board under four different illuminations (white, red, green and blue). Subsequently, an improved system including a ring light, an objective lens, and a monochromatic camera was set up to acquire higher quality images. The obtained images can be divided into three main components: the PCB itself (i.e., the background), the reflections induced by unsoldered positions or screw holes and the solder joints. Non-uniform illumination is corrected by estimating the background using a morphological opening and subtraction from the input image. Image sharpening is applied in order to prevent error pixels in the subsequent segmentation. The intensity thresholds which divide the main components are obtained from the multimodal histogram using three probability density functions. Determining the intersections delivers proper thresholds for the segmentation. Remaining edge gradients produces small error areas which are removed by another morphological opening. For quantitative analysis of the segmentation results, the dice coefficient is used. Results: The obtained PCB images show a significant gradient in all RGB channels, resulting from ambient light. Using different lightings and color channels 12 images of a single PCB are available. A visual inspection and the investigation of 27 specific points show the best differentiation between those points using a red lighting and a green color channel. Estimating two thresholds from analyzing the multimodal histogram of the corrected images and using them for segmentation precisely extracts the solder joints. The comparison of the results to manually segmented images yield high sensitivity and specificity values. Analyzing the overall result delivers a Dice coefficient of 0.89 which varies for single object segmentations between 0.96 for a good segmented solder joints and 0.25 for single negative outliers. Conclusion: Our results demonstrate that the presented optical imaging system and the developed algorithm can robustly detect solder joints on printed circuit boards. Future work will comprise a modified lighting system which allows for more precise segmentation results using structure analysis.

Keywords: printed circuit board jet-printing, inspection, segmentation, solder paste detection

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72 A Comparative Study of Efficacy and Safety of Salicylic Acid, Trichloroacetic Acid and Glycolic Acid in Various Facial Melanosis

Authors: Shivani Dhande, Sanjiv Choudhary, Adarshlata Singh

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Introduction: Chemical peeling is a popular, relatively inexpensive day procedure and generally safe method for treatment of pigmentary skin disorders and for skin rejuvenation. Chemical peels are classified by the depth of action into superficial, medium, and deep peels.Various facial pigmentary conditions have significant impact on quality of life causing psychological stress, necessitating its safe and effective treatment.Aim & Objectives:To compare the efficacy of Salicylic acid, Trichloroaceticacid & Glycolic Acid in facial melanosis(melasma,photomelanosis& post acne pigmentation).To study the side effects of above mentioned peeling agents. Method and Materials:It was a randomized parallel control single blind study consisting of total of 36 cases, 12 cases each of melasma, photo melanosis and post acne pigmentation within age group 20-50 years having fitzpatrick’s skin type4. Woods lamp examination was done to confirm the type of melasma.Patients with keloidal tendency, active herpes infection or past history of hypersensitivity to salicylic acid, trichloroaceticand glycolic acid as well aspatients on systemic isotretinoin were excluded.Clinical photographs at the beginning of therapy and then serially, were taken to assess the clinical response. Prior to application a written informed consent was obtained. A post auricular test peel was performed. Patients were divided into 3 groups, containing 12 patients each of melasma, photomelanosis and post acnepigmentation.All the three peels SA peel 20% (done once in 2 weeks), GA peel 50% (done once in 3 weeks) and TCA 15% (done once in 3 weeks) were used with total six settings for each patient. Before application of peel patients were counseled to wash the face with soap and water. Then face was dried and cleaned with spirit and acetone to remove all cutaneous oils. GA, TCA, SA were applied with cotton buds/gauze withmild strokes. After a contact period off 5-10mins neutralization was done with cold water. Post peel topical sunscreen application was mandatory. MASI was used pre and post treatment to assess melasma. Investigator’s global improvement scale- overall hyperpigmentation (4-significant, 3-moderate, 2-mild, 1-minimal, 0-no change ) and Patient’s satisfaction grading scale (>70%- excellent response, 50-70%- good response, <50%- average response) was used to assess improvement in all the three facial melanosis.Results:In our study of 12 patients of melasma, 4 (33.33%)patients showed excellent results;3 (25%) with GAand 1(8.33%) of TCA.Good response was seen in 4 (33.33%) patients;1(8.33%) each for GA & SA and 2(16.66%) for TCA.Poor response was seen in 4(33.33%) patients;1(8.33%) for TCA and 3 (25%) for SA.Of 12 patients of photomelanosis, excellent resultswas seen in 3(25%)patients of TCA. Good response was seen in 4 (33.33%) patients, 1(8.33%) each of TCA &SA and 2(16.66%) of GA.Poor responsewas seen in 5(41.66%) patients;3 (25%) for SA and 2(16.66%) of GA.Of 12 patients of post acne pigmentation, excellent responsein 3 (25%) patients;2(16.66%) of SA and 1(8.33%) of TCA.Good responsewas seen in 5(41.66%) patients;2(16.66%) of SA and GA and1(8.33%) of TCA.Poor response was seen in 4 (33.33%) patients; 2 (16.66%) for SA and TCA both. No major side effects in the form of scarring or persistant pigmentation was seen. Transient blackening of skin with burning sensation was seen in cases treated with TCA and SA. Post procedural itching and redness was noted with GA peel. Conclusion- In our study GA(50%),TCA(15%) & SA(20%) peels showed excellent response in melasma, photomelanosis and post-acne pigmentation respectively.All the 3 peeling agents were well tolerated without any significant side-effects in the above specified concentrations.

Keywords: facial melanosis, gycolic acid, salicylic acid, trichloroacetic acid

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71 Participation of Titanium Influencing the Petrological Assemblage of Mafic Dyke: Salem, South India

Authors: Ayoti Banerjee, Meenakshi Banerjee

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The study of metamorphic reaction textures is important in contributing to our understanding of the evolution of metamorphic terranes. Where preserved, they provide information on changes in the P-T conditions during the metamorphic history of the rock, and thus allow us to speculate on the P-T-t evolution of the terrane. Mafic dykes have attracted the attention of petrologists because they act as window to mantle. This rock represents a mafic dyke of doleritic composition. It is fine to medium grained in which clinopyroxene are enclosed by the lath shaped plagioclase grains to form spectacular ophitic texture. At places, sub ophitic texture was also observed. Grains of pyroxene and plagioclase show very less deformation typically plagioclase showing deformed lamella along with plagioclase-clinopyroxene-phyric granoblastic fabric within a groundmass of feldspar microphenocrysts and Fe–Ti oxides. Both normal and reverse zoning were noted in the plagioclase laths. The clinopyroxene grains contain exsolved phases such as orthopyroxene, plagioclase, magnetite, ilmenite along the cleavage traces and the orthopyroxene lamella form granules in the periphery of the clinopyroxene grains. Garnet corona also develops preferentially around plagioclase at the contact of clinopyroxene, ilmenite or magnetite. Tiny quartz and K-fs grains showed symplectic intergrowth with garnet at a few places. The product quartz formed along with garnet rims the coronal garnet and the reacting clinopyroxene. Thin amphibole corona formed along the periphery of deformed plagioclase and clinopyroxene occur as patches over the magmatic minerals. The amphibole coronas cannot be assigned to a late magmatic stage and are interpreted as reactive being restricted to the contact between clinopyroxene and plagioclase, thus postdating the crystallization of both. The amphibole and garnet do not share grain boundary in the entire rock and is thus pointing towards simultaneous crystallization. Olivine is absent. Spectacular myrmekitic growth of orthoclase and quartz rimming the plagioclase is consistent with the potash metasomatic effects that is also found in other rocks of this region. These textural features are consistent with a phase of fluid induced metamorphism (retrogression). But the appearance of coronal garnet and amphibole exclusive of each other reflects the participation if Ti as the prime reason. Presence of Ti as a reactant phase is a must for amphibole forming reactions whereas it is not so in case of garnet forming reactions although the reactants are the same plagioclase and clinopyroxene in both cases. These findings are well validated by petrographical and textural analysis. In order to obtain balanced chemical reactions that explain formation of amphibole and garnet in the mafic dyke rocks a matrix operation technique called Singular Value Decomposition (SVD) was adopted utilizing the measured chemical compositions of the minerals. The computer program C-Space was used for this purpose and the required compositional matrix. Data fed to C-Space was after doing cation-calculation of the oxide percentages obtained from EPMA analysis. The Garnet-Clinopyroxene geothermometer yielded a temperature of 650 degrees Celsius. The Garnet-Clinopyroxene-Plagioclase geobarometer and Al-in amphibole yielded roughly 7.5 kbar pressure.

Keywords: corona, dolerite, geothermometer, metasomatism, metamorphic reaction texture, retrogression

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70 Mental Health Promotion for Children of Mentally Ill Parents in Schools. Assessment and Promotion of Teacher Mental Health Literacy in Order to Promote Child Related Mental Health (Teacher-MHL)

Authors: Dirk Bruland, Paulo Pinheiro, Ullrich Bauer

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Introduction: Over 3 million children, about one quarter of all students, experience at least one parent with mental disorder in Germany every year. Children of mentally-ill parents are at considerably higher risk of developing serious mental health problems. The different burden patterns and coping attempts often become manifest in children's school lives. In this context, schools can have an important protective function, but can also create risk potentials. In reference to Jorm, pupil-related teachers’ mental health literacy (Teacher-MHL) includes the ability to recognize change behaviour, the knowledge of risk factors, the implementation of first aid intervention, and seeking professional help (teacher as gatekeeper). Although teachers’ knowledge and increased awareness of this topic is essential, the literature provides little information on the extent of teachers' abilities. As part of a German-wide research consortium on health literacy, this project, launched in March for 3 years, will conduct evidence-based mental health literacy research. The primary objective is to measure Teacher-MHL in the context of pupil-related psychosocial factors at primary and secondary schools (grades 5 & 6), while also focussing on children’s social living conditions. Methods: (1) A systematic literature review in different databases to identify papers with regard to Teacher-MHL (completed). (2) Based on these results, an interview guide was developed. This research step includes a qualitative pre-study to inductively survey the general profiles of teachers (n=24). The evaluation will be presented on the conference. (3) These findings will be translated into a quantitative teacher survey (n=2500) in order to assess the extent of socio-analytical skills of teachers as well as in relation to institutional and individual characteristics. (4) Based on results 1 – 3, developing a training program for teachers. Results: The review highlights a lack of information for Teacher-MHL and their skills, especially related to high-risk-groups like children of mentally ill parents. The literature is limited to a few studies only. According to these, teacher are not good at identifying burdened children and if they identify those children they do not know how to handle the situations in school. They are not sufficiently trained to deal with these children, especially there are great uncertainties in dealing with the teaching situation. Institutional means and resources are missing as well. Such a mismatch can result in insufficient support and use of opportunities for children at risk. First impressions from the interviews confirm these results and allow a greater insight in the everyday school-life according to critical life events in families. Conclusions: For the first time schools will be addressed as a setting where children are especially "accessible" for measures of health promotion. Addressing Teacher-MHL gives reason to expect high effectiveness. Targeting professionals' abilities for dealing with this high-risk-group leads to a discharge for teacher themselves to handle those situations and increases school health promotion. In view of the fact that only 10-30% of such high-risk families accept offers of therapy and assistance, this will be the first primary preventive and health-promoting approach to protect the health of a yet unaffected, but particularly burdened, high-risk group.

Keywords: children of mentally ill parents, health promotion, mental health literacy, school

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69 Mixed Mode Fracture Analyses Using Finite Element Method of Edge Cracked Heavy Annulus Pulley

Authors: Bijit Kalita, K. V. N. Surendra

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The pulley works under both compressive loading due to contacting belt in tension and central torque due to cause rotation. In a power transmission system, the belt pulley assemblies offer a contact problem in the form of two mating cylindrical parts. In this work, we modeled a pulley as a heavy two-dimensional circular disk. Stress analysis due to contact loading in the pulley mechanism is performed. Finite element analysis (FEA) is conducted for a pulley to investigate the stresses experienced on its inner and outer periphery. In most of the heavy-duty applications, most frequently used mechanisms to transmit power in applications such as automotive engines, industrial machines, etc. is Belt Drive. Usually, very heavy circular disks are used as pulleys. A pulley could be entitled as a drum and may have a groove between two flanges around the circumference. A rope, belt, cable or chain can be the driving element of a pulley system that runs over the pulley inside the groove. A pulley is experienced by normal and shear tractions on its contact region in the process of motion transmission. The region may be belt-pulley contact surface or pulley-shaft contact surface. In 1895, Hertz solved the elastic contact problem for point contact and line contact of an ideal smooth object. Afterward, this hypothesis is generally utilized for computing the actual contact zone. Detailed stress analysis in such contact region of such pulleys is quite necessary to prevent early failure. In this paper, the results of the finite element analyses carried out on the compressed disk of a belt pulley arrangement using fracture mechanics concepts are shown. Based on the literature on contact stress problem induced in the wide field of applications, generated stress distribution on the shaft-pulley and belt-pulley interfaces due to the application of high-tension and torque was evaluated in this study using FEA concepts. Finally, the results obtained from ANSYS (APDL) were compared with the Hertzian contact theory. The study is mainly focused on the fatigue life estimation of a rotating part as a component of an engine assembly using the most famous Paris equation. Digital Image Correlation (DIC) analyses have been performed using the open-source software. From the displacement computed using the images acquired at a minimum and maximum force, displacement field amplitude is computed. From these fields, the crack path is defined and stress intensity factors and crack tip position are extracted. A non-linear least-squares projection is used for the purpose of the estimation of fatigue crack growth. Further study will be extended for the various application of rotating machinery such as rotating flywheel disk, jet engine, compressor disk, roller disk cutter etc., where Stress Intensity Factor (SIF) calculation plays a significant role on the accuracy and reliability of a safe design. Additionally, this study will be progressed to predict crack propagation in the pulley using maximum tangential stress (MTS) criteria for mixed mode fracture.

Keywords: crack-tip deformations, contact stress, stress concentration, stress intensity factor

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68 Pluripotent Stem Cells as Therapeutic Tools for Limbal Stem Cell Deficiencies and Drug Testing

Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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67 Fresh Amnion Membrane Grafting for the Regeneration of Skin in Full Thickness Burn in Newborn - Case Report

Authors: Priyanka Yadav, Umesh Bnasal, Yashvinder Kumar

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The placenta is an important structure that provides oxygen and nutrients to the growing fetus in utero. It is usually thrown away after birth, but it has a therapeutic role in the regeneration of tissue. It is covered by the amniotic membrane, which can be easily separated into the amnion layer and the chorion layer—the amnion layer act as a biofilm for the healing of burn wound and non-healing ulcers. The freshly collected membrane has stem cells, cytokines, growth factors, and anti-inflammatory properties, which act as a biofilm for the healing of wounds. It functions as a barrier and prevents heat and water loss and also protects from bacterial contamination, thus supporting the healing process. The application of Amnion membranes has been successfully used for wound and reconstructive purposes for decades. It is a very cheap and easy process and has shown superior results to allograft and xenograft. However, there are very few case reports of amnion membrane grafting in newborns; we intend to highlight its therapeutic importance in burn injuries in newborns. We present a case of 9 days old male neonate who presented to the neonatal unit of Maulana Azad Medical College with a complaint of fluid-filled blisters and burns wound on the body for six days. He was born outside the hospital at 38 weeks of gestation to a 24-year-old primigravida mother by vaginal delivery. The presentation was cephalic and the amniotic fluid was clear. His birth weight was 2800 gm and APGAR scores were 7 and 8 at 1 and 5 minutes, respectively. His anthropometry was appropriate for gestational age. He developed respiratory distress after birth requiring oxygen support by nasal prongs for three days. On the day of life three, he developed blisters on his body, starting from than face then over the back and perineal region. At a presentation on the day of life nine, he had blisters and necrotic wound on the right side of the face, back, right shoulder and genitalia, affecting 60% of body surface area with full-thickness loss of skin. He was started on intravenous antibiotics and fluid therapy. Pus culture grew Pseudomonas aeuroginosa, for which culture-specific antibiotics were started. Plastic surgery reference was taken and regular wound dressing was done with antiseptics. He had a storming course during the hospital stay. On the day of life 35 when the baby was hemodynamically stable, amnion membrane grafting was done on the wound site; for the grafting, fresh amnion membrane was removed under sterile conditions from the placenta obtained by caesarean section. It was then transported to the plastic surgery unit in half an hour in a sterile fluid where the graft was applied over the infant’s wound. The amnion membrane grafting was done twice in two weeks for covering the whole wound area. After successful uptake of amnion membrane, skin from the thigh region was autografted over the whole wound area by Meek technique in a single setting. The uptake of autograft was excellent and most of the areas were healed. In some areas, there was patchy regeneration of skin so dressing was continued. The infant was discharged after three months of hospital stay and was later followed up in the plastic surgery unit of the hospital.

Keywords: amnion membrane grafting, autograft, meek technique, newborn, regeneration of skin

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66 Integrating Non-Psychoactive Phytocannabinoids and Their Cyclodextrin Inclusion Complexes into the Treatment of Glioblastoma

Authors: Kyriaki Hatziagapiou, Konstantinos Bethanis, Olti Nikola, Elias Christoforides, Eleni Koniari, Eleni Kakouri, George Lambrou, Christina Kanaka-Gantenbein

Abstract:

Glioblastoma multiforme (GBM) remains a serious health challenge, as current therapeutic modalities continue to yield unsatisfactory results, with the average survival rarely exceeding 1-2 years. Natural compounds still provide some of the most promising approaches for discovering new drugs. The non-psychotropic cannabidiol (CBD) deriving from Cannabis sativa L. provides such promise. CBD is endowed with anticancer, antioxidant, and genoprotective properties as established in vitro and in in vivo experiments. CBD’s selectivity towards cancer cells and its safe profile suggest its usage in cancer therapies. However, the bioavailability of oral CBD is low due to poor aqueous solubility, erratic gastrointestinal absorption, and significant first-pass metabolism, hampering its therapeutic potential and resulting in a variable pharmacokinetic profile. In this context, CBD can take great advantage of nanomedicine-based formulation strategies. Cyclodextrins (CDs) are cyclic oligosaccharides used in the pharmaceutical industry to incorporate apolar molecules inside their hydrophobic cavity, increasing their stability, water solubility, and bioavailability or decreasing their side effects. CBD-inclusion complexes with CDs could be a good strategy to improve its properties, like solubility and stability to harness its full therapeutic potential. The current research aims to study the potential cytotoxic effect of CBD and CBD-CDs complexes CBD-RMβCD (randomly methylated β-cyclodextrin) and CBD-HPβCD (hydroxypropyl-b-CD) on the A172 glioblastoma cell line. CBD is diluted in 10% DMSO, and CBD/CDs solutions are prepared by mixing solid CBD, solid CDs, and dH2O. For the biological assays, A172 cells are incubated at a range of concentrations of CBD, CBD-RMβCD and CBD-HPβCD, RMβCD, and HPβCD (0,03125-4 mg/ml) at 24, 48, and 72 hours. Analysis of cell viability after incubation with the compounds is performed with Alamar Blue viability assay. CBD’s dilution to DMSO 10% was inadequate, as crystals are observed; thus cytotoxicity experiments are not assessed. CBD’s solubility is enhanced in the presence of both CDs. CBD/CDs exert significant cytotoxicity in a dose and time-dependent manner (p < 0.005 for exposed cells to any concentration at 48, 72, and 96 hours versus cells not exposed); as their concentration and time of exposure increases, the reduction of resazurin to resofurin decreases, indicating a reduction in cell viability. The cytotoxic effect is more pronounced in cells exposed to CBD-HPβCD for all concentrations and time-points. RMβCD and HPβCD at the highest concentration of 4 mg/ml also exerted antitumor action per se since manifesting cell growth inhibition. The results of our study could afford the basis of research regarding the use of natural products and their inclusion complexes as anticancer agents and the shift to targeted therapy with higher efficacy and limited toxicity. Acknowledgments: The research is partly funded by ΙΚΥ (State Scholarships Foundation) – Post-doc Scholarships-Partnership Agreement 2014-2020.

Keywords: cannabidiol, cyclodextrins, glioblastoma, hydroxypropyl-b-Cyclodextrin, randomly-methylated-β-cyclodextrin

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65 Blood Lipid Management: Combined Treatment with Hydrotherapy and Ozone Bubbles Bursting in Water

Authors: M. M. Wickramasinghe

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Cholesterol and triglycerides are lipids, mainly essential to maintain the cellular structure of the human body. Cholesterol is also important for hormone production, vitamin D production, proper digestion functions, and strengthening the immune system. Excess fats in the blood circulation, known as hyperlipidemia, become harmful leading to arterial clogging and causing atherosclerosis. Aim of this research is to develop a treatment protocol to efficiently break down and maintain circulatory lipids by improving blood circulation without strenuous physical exercises while immersed in a tub of water. To achieve the target of strong exercise effect, this method involves generating powerful ozone bubbles to spin, collide, and burst in the water. Powerful emission of air into water is capable of transferring locked energy of the water molecules and releasing energy. This method involves water and air-based impact generated by pumping ozone at the speed of 46 lts/sec with a concentration of 0.03-0.05 ppt according to safety standards of The Federal Institute for Drugs and Medical Devices, BfArM, Germany. The direct impact of ozone bubbles on the muscular system and skin becomes the main target and is capable of increasing the heart rate while immersed in water. A total time duration of 20 minutes is adequate to exert a strong exercise effect, improve blood circulation, and stimulate the nervous and endocrine systems. Unstable ozone breakdown into oxygen release onto the surface of the water giving additional benefits and supplying high-quality air rich in oxygen required to maintain efficient metabolic functions. The breathing technique was introduced to improve the efficiency of lung functions and benefit the air exchange mechanism. The temperature of the water is maintained at 39c to 40c to support arterial dilation and enzyme functions and efficiently improve blood circulation to the vital organs. The buoyancy of water and natural hydrostatic pressure release the tension of the body weight and relax the mind and body. Sufficient hydration (3lts of water per day) is an essential requirement to transport nutrients and remove waste byproducts to process through the liver, kidney, and skin. Proper nutritional intake is an added advantage to optimize the efficiency of this method which aids in a fast recovery process. Within 20-30 days of daily treatment, triglycerides, low-density lipoproteins (LDL), and total cholesterol reduction were observed in patients with abnormal levels of lipid profile. Borderline patients were cleared within 10–15 days of treatment. This is a highly efficient system that provides many benefits and is able to achieve a successful reduction of triglycerides, LDL, and total cholesterol within a short period of time. Supported by proper hydration and nutritional balance, this system of natural treatment maintains healthy levels of lipids in the blood and avoids the risk of cerebral stroke, high blood pressure, and heart attacks.

Keywords: atherosclerosis, cholesterol, hydrotherapy, hyperlipidemia, lipid management, ozone therapy, triglycerides

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64 Understanding Responses of the Bee Community to an Urbanizing Landscape in Bengaluru, South India

Authors: Chethana V. Casiker, Jagadishakumara B., Sunil G. M., Chaithra K., M. Soubadra Devy

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A majority of the world’s food crops depends on insects for pollination, among which bees are the most dominant taxon. Bees pollinate vegetables, fruits and oilseeds which are rich in essential micronutrients. Besides being a prerequisite for a nutritionally secure diet, agrarian economies such as India depend heavily on pollination for good yield and quality of the product. As cities all over the world expand rapidly, large tracts of green spaces are being built up. This, along with high usage of agricultural chemicals has reduced floral diversity and shrunk bee habitats. Indeed, pollinator decline is being reported from various parts of the world. Further, the FAO has reported a huge increase in the area of land under cultivation of pollinator-dependent crops. In the light of increasing demand for pollination and disappearing natural habitats, it is critical to understand whether and how urban spaces can support pollinators. To this end, this study investigates the influence of landscape and local habitat quality on bee community dynamics. To capture the dynamics of expanding cityscapes, the study employs a space for time substitution, wherein a transect along the gradient of urbanization substitutes a timeframe of increasing urbanization. This will help understand how pollinators would respond to changes induced by increasing intensity of urbanization in the future. Bengaluru, one of the fastest growing cities of Southern India, is an excellent site to study impacts associated with urbanization. With sites moving away from the Bengaluru’s centre and towards its peripheries, this study captures the changes in bee species diversity and richness along a gradient of urbanization. Bees were sampled under different land use types as well as in different types of vegetation, including plantations, croplands, fallow land, parks, lake embankments, and private gardens. The relationship between bee community metrics and key drivers such as a percentage of built-up area, land use practices, and floral resources was examined. Additionally, data collected using questionnaire interviews were used to understand people’s perceptions towards and level of dependence on pollinators. Our results showed that urban areas are capable of supporting bees. In fact, a greater diversity of bees was recorded in urban sites compared to adjoining rural areas. This suggests that bees are able to seek out patchy resources and survive in small fragments of habitat. Bee abundance and species richness correlated positively with floral abundance and richness, indicating the role of vegetation in providing forage and nesting sites which are crucial to their survival. Bee numbers were seen to decrease with increase in built-up area demonstrating that impervious surfaces could act as deterrents. Findings from this study challenge the popular notion of cities being biodiversity-bare spaces. There is indeed scope for conserving bees in urban landscapes, provided that there are city-scale planning and local initiative. Bee conservation can go hand in hand with efforts such as urban gardening and terrace farming that could help cities urbanize sustainably.

Keywords: bee, landscape ecology, urbanization, urban pollination

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63 An Elasto-Viscoplastic Constitutive Model for Unsaturated Soils: Numerical Implementation and Validation

Authors: Maria Lazari, Lorenzo Sanavia

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Mechanics of unsaturated soils has been an active field of research in the last decades. Efficient constitutive models that take into account the partial saturation of soil are necessary to solve a number of engineering problems e.g. instability of slopes and cuts due to heavy rainfalls. A large number of constitutive models can now be found in the literature that considers fundamental issues associated with the unsaturated soil behaviour, like the volume change and shear strength behaviour with suction or saturation changes. Partially saturated soils may either expand or collapse upon wetting depending on the stress level, and it is also possible that a soil might experience a reversal in the volumetric behaviour during wetting. Shear strength of soils also changes dramatically with changes in the degree of saturation, and a related engineering problem is slope failures caused by rainfall. There are several states of the art reviews over the last years for studying the topic, usually providing a thorough discussion of the stress state, the advantages, and disadvantages of specific constitutive models as well as the latest developments in the area of unsaturated soil modelling. However, only a few studies focused on the coupling between partial saturation states and time effects on the behaviour of geomaterials. Rate dependency is experimentally observed in the mechanical response of granular materials, and a viscoplastic constitutive model is capable of reproducing creep and relaxation processes. Therefore, in this work an elasto-viscoplastic constitutive model for unsaturated soils is proposed and validated on the basis of experimental data. The model constitutes an extension of an existing elastoplastic strain-hardening constitutive model capable of capturing the behaviour of variably saturated soils, based on energy conjugated stress variables in the framework of superposed continua. The purpose was to develop a model able to deal with possible mechanical instabilities within a consistent energy framework. The model shares the same conceptual structure of the elastoplastic laws proposed to deal with bonded geomaterials subject to weathering or diagenesis and is capable of modelling several kinds of instabilities induced by the loss of hydraulic bonding contributions. The novelty of the proposed formulation is enhanced with the incorporation of density dependent stiffness and hardening coefficients in order to allow the modeling of the pycnotropy behaviour of granular materials with a single set of material constants. The model has been implemented in the commercial FE platform PLAXIS, widely used in Europe for advanced geotechnical design. The algorithmic strategies adopted for the stress-point algorithm had to be revised to take into account the different approach adopted by PLAXIS developers in the solution of the discrete non-linear equilibrium equations. An extensive comparison between models with a series of experimental data reported by different authors is presented to validate the model and illustrate the capability of the newly developed model. After the validation, the effectiveness of the viscoplastic model is displayed by numerical simulations of a partially saturated slope failure of the laboratory scale and the effect of viscosity and degree of saturation on slope’s stability is discussed.

Keywords: PLAXIS software, slope, unsaturated soils, Viscoplasticity

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62 LncRNA-miRNA-mRNA Networks Associated with BCR-ABL T315I Mutation in Chronic Myeloid Leukemia

Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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61 Erectile Dysfunction in A Middle Aged Man 6 Years After Bariatric Surgery: A Case Report

Authors: Thaminda Liyanage, Chamila Shamika Kurukulasuriya

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Introduction: Morbid obesity has been successfully treated with bariatric surgery for over 60 years. Although operative procedures have improved and associated complications have reduced substantially, surgery still carries the risk of post-operative malabsorption, malnutrition and a range of gastrointestinal disorders. Overweight by itself can impair libido in both sexes and cause erectile dysfunction in males by inducing a state of hypogonadotropic hypogonadism, proportional to the degree of obesity. Impact of weight reduction on libido and sexual activity remains controversial, however it is broadly accepted that weight loss improves sexual drive. Zinc deficiency, subsequent to malabsorption, may lead to impaired testosterone synthesis in men while excessive and/or rapid weight loss in females may result in reversible amenorrhoea leading to sub-fertility. Methods: We describe a 37 year old male, 6 years post Roux-en-Y gastric bypass surgery, who presented with erectile dysfunction, loss of libido, worsening fatigue and generalized weakness for 4 months. He also complained of constipation and frequent muscle cramps but denied having headache, vomiting or visual disturbances. Patient had lost 38 kg of body weight post gastric bypass surgery over four years {135kg (BMI 42.6 kg/m2) to 97 kg (BMI 30.6 kg/m2)} and the weight had been stable for past two years. He had no recognised co-morbidities at the time of the surgery and noted marked improvement in general wellbeing, physical fitness and psychological confident post surgery, up until four months before presentation. Clinical examination revealed dry pale skin with normal body hair distribution, no thyroid nodules or goitre, normal size testicles and normal neurological examination with no visual field defects or diplopia. He had low serum testosterone, follicular stimulating hormone (FSH), luteinizing hormone (LH), T3, T4, thyroid stimulating hormone (TSH), insulin like growth factor 1 (IGF-1) and 24-hour urine cortisol levels. Serum cortisol demonstrated an appropriate rise to ACTH stimulation test but growth hormone (GH) failed increase on insulin tolerance test. Other biochemical and haematological studies were normal, except for low zinc and folate with minimally raised liver enzymes. MRI scan of the head confirmed a solid pituitary mass with no mass effect on optic chiasm. Results: In this patient clinical, biochemical and radiological findings were consistent with anterior pituitary dysfunction. However, there were no features of raised intracranial pressure or neurological compromise. He was commenced on appropriate home replacement therapy and referred for neurosurgical evaluation. Patient reported marked improvement in his symptoms, specially libido and erectile dysfunction, on subsequent follow up visits. Conclusion: Sexual dysfunction coupled with non specific constitutional symptoms has multiple aetiologies. Clinical symptoms out of proportion to nutritional deficiencies post bariatric surgery should be thoroughly investigated. Close long term follow up is crucial for overall success.

Keywords: obesity, bariatric surgery, erectile dysfunction, loss of libido

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60 Green Building Risks: Limits on Environmental and Health Quality Metrics for Contractors

Authors: Erica Cochran Hameen, Bobuchi Ken-Opurum, Mounica Guturu

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The United Stated (U.S.) populous spends the majority of their time indoors in spaces where building codes and voluntary sustainability standards provide clear Indoor Environmental Quality (IEQ) metrics. The existing sustainable building standards and codes are aimed towards improving IEQ, health of occupants, and reducing the negative impacts of buildings on the environment. While they address the post-occupancy stage of buildings, there are fewer standards on the pre-occupancy stage thereby placing a large labor population in environments much less regulated. Construction personnel are often exposed to a variety of uncomfortable and unhealthy elements while on construction sites, primarily thermal, visual, acoustic, and air quality related. Construction site power generators, equipment, and machinery generate on average 9 decibels (dBA) above the U.S. OSHA regulations, creating uncomfortable noise levels. Research has shown that frequent exposure to high noise levels leads to chronic physiological issues and increases noise induced stress, yet beyond OSHA no other metric focuses directly on the impacts of noise on contractors’ well-being. Research has also associated natural light with higher productivity and attention span, and lower cases of fatigue in construction workers. However, daylight is not always available as construction workers often perform tasks in cramped spaces, dark areas, or at nighttime. In these instances, the use of artificial light is necessary, yet lighting standards for use during lengthy tasks and arduous activities is not specified. Additionally, ambient air, contaminants, and material off-gassing expelled at construction sites are one of the causes of serious health effects in construction workers. Coupled with extreme hot and cold temperatures for different climate zones, health and productivity can be seriously compromised. This research evaluates the impact of existing green building metrics on construction and risk management, by analyzing two codes and nine standards including LEED, WELL, and BREAM. These metrics were chosen based on the relevance to the U.S. construction industry. This research determined that less than 20% of the sustainability context within the standards and codes (texts) are related to the pre-occupancy building sector. The research also investigated the impact of construction personnel’s health and well-being on construction management through two surveys of project managers and on-site contractors’ perception of their work environment on productivity. To fully understand the risks of limited Environmental and Health Quality metrics for contractors (EHQ) this research evaluated the connection between EHQ factors such as inefficient lighting, on construction workers and investigated the correlation between various site coping strategies for comfort and productivity. Outcomes from this research are three-pronged. The first includes fostering a discussion about the existing conditions of EQH elements, i.e. thermal, lighting, ergonomic, acoustic, and air quality on the construction labor force. The second identifies gaps in sustainability standards and codes during the pre-occupancy stage of building construction from ground-breaking to substantial completion. The third identifies opportunities for improvements and mitigation strategies to improve EQH such as increased monitoring of effects on productivity and health of contractors and increased inclusion of the pre-occupancy stage in green building standards.

Keywords: construction contractors, health and well-being, environmental quality, risk management

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59 Socio-Economic Determinants of Physical Activity of Non-Manual Workers, Including the Early Senior Group, from the City of Wroclaw in Poland

Authors: Daniel Puciato, Piotr Oleśniewicz, Julita Markiewicz-Patkowska, Krzysztof Widawski, Michał Rozpara, Władysław Mynarski, Agnieszka Gawlik, Małgorzata Dębska, Soňa Jandová

Abstract:

Physical activity as a part of people’s everyday life reduces the risk of many diseases, including those induced by lifestyle, e.g. obesity, type 2 diabetes, osteoporosis, coronary heart disease, degenerative arthritis, and certain types of cancer. That refers particularly to professionally active people, including the early senior group working on non-manual positions. The aim of the study is to evaluate the relationship between physical activity and the socio-economic status of non-manual workers from Wroclaw—one of the biggest cities in Poland, a model setting for such investigations in this part of Europe. The crucial problem in the research is to find out the percentage of respondents who meet the health-related recommendations of the World Health Organization (WHO) concerning the volume, frequency, and intensity of physical activity, as well as to establish if the most important socio-economic factors, such as gender, age, education, marital status, per capita income, savings and debt, determine the compliance with the WHO physical activity recommendations. During the research, conducted in 2013, 1,170 people (611 women and 559 men) aged 21–60 years were examined. A diagnostic poll method was applied to collect the data. Physical activity was measured with the use of the short form of the International Physical Activity Questionnaire with extended socio-demographic questions, i.e. concerning gender, age, education, marital status, income, savings or debts. To evaluate the relationship between physical activity and selected socio-economic factors, logistic regression was used (odds ratio statistics). Statistical inference was conducted on the adopted ex ante probability level of p<0.05. The majority of respondents met the volume of physical effort recommended for health benefits. It was particularly noticeable in the case of the examined men. The probability of compliance with the WHO physical activity recommendations was highest for workers aged 21–30 years with secondary or higher education who were single, received highest incomes and had savings. The results indicate the relations between physical activity and socio-economic status in the examined women and men. People with lower socio-economic status (e.g. manual workers) are physically active primarily at work, whereas those better educated and wealthier implement physical effort primarily in their leisure time. Among the investigated subjects, the youngest group of non-manual workers have the best chances to meet the WHO standards of physical activity. The study also confirms that secondary education has a positive effect on the public awareness on the role of physical activity in human life. In general, the analysis of the research indicates that there is a relationship between physical activity and some socio-economic factors of the respondents, such as gender, age, education, marital status, income per capita, and the possession of savings. Although the obtained results cannot be applied for the general population, they show some important trends that will be verified in subsequent studies conducted by the authors of the paper.

Keywords: IPAQ, nonmanual workers, physical activity, socioeconomic factors, WHO

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58 Glucose Uptake Rate of Insulin-Resistant Human Liver Carcinoma Cells (IR/HepG2) by Flavonoids from Enicostema littorale via IR/IRS1/AKT Pathway

Authors: Priyanka Mokashi, Aparna Khanna, Nancy Pandita

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Diabetes mellitus is a chronic metabolic disorder which will be the 7th leading cause of death by 2030. The current line of treatment for the diabetes mellitus is oral antidiabetic drugs (biguanides, sulfonylureas, meglitinides, thiazolidinediones and alpha-glycosidase inhibitors) and insulin therapy depending upon the type 1 or type 2 diabetes mellitus. But, these treatments have their disadvantages, ranging from the developing of resistance to the drugs and adverse effects caused by them. Alternative to these synthetic agents, natural products provides a new insight for the development of more efficient and safe drugs due to their therapeutic values. Enicostema littorale blume (A. Raynal) is a traditional Indian plant belongs to the Gentianaceae family. It is widely distributed in Asia, Africa, and South America. There are few reports on Swrtiamarin, major component of this plant for its antidiabetic activity. However, the antidiabetic activity of flavonoids from E. littorale and their mechanism of action have not yet been elucidated. Flavonoids have a positive relationship with disease prevention and can act on various molecular targets and regulate different signaling pathways in pancreatic β-cells, adipocytes, hepatocytes and skeletal myofibers. They may exert beneficial effects in diabetes by (i) improving hyperglycemia through regulation of glucose metabolism in hepatocytes; (ii) enhancing insulin secretion and reducing apoptosis and promoting proliferation of pancreatic β-cells; (iii) increasing glucose uptake in hepatocytes, skeletal muscle and white adipose tissue (iv) reducing insulin resistance, inflammation and oxidative stress. Therefore, we have isolated four flavonoid rich fractions, Fraction A (FA), Fraction B (FB), Fraction C (FC), Fraction D (FD) from crude alcoholic hot (AH) extract from E. littorale, identified by LC/MS. Total eight flavonoids were identified on the basis of fragmentation pattern. Flavonoid FA showed the presence of swertisin, isovitexin, and saponarin; FB showed genkwanin, quercetin, isovitexin, FC showed apigenin, swertisin, quercetin, 5-O-glucosylswertisin and 5-O-glucosylisoswertisin whereas FD showed the presence of swertisin. Further, these fractions were assessed for their antidiabetic activity on stimulating glucose uptake in insulin-resistant HepG2 cell line model (IR/HepG2). The results showed that FD containing C-glycoside Swertisin has significantly increased the glucose uptake rate of IR/HepG2 cells at the concentration of 10 µg/ml as compared to positive control Metformin (0.5mM) which was determined by glucose oxidase- peroxidase method. It has been reported that enhancement of glucose uptake of cells occurs due the translocation of Glut4 vesicles to cell membrane through IR/IRS1/AKT pathway. Therefore, we have studied expressions of three genes IRS1, AKT and Glut4 by real-time PCR to evaluate whether they follow the same pathway or not. It was seen that the glucose uptake rate has increased in FD treated IR/HepG2 cells due to the activation of insulin receptor substrate-1 (IRS1) followed by protein kinase B (AKT) through phosphoinositide 3-kinase (PI3K) leading to translocation of Glut 4 vesicles to cell membrane, thereby enhancing glucose uptake and insulin sensitivity of insulin resistant HepG2 cells. Hence, the up-regulation indicated the mechanism of action through which FD (Swertisin) acts as antidiabetic candidate in the treatment of type 2 diabetes mellitus.

Keywords: E. littorale, glucose transporter, glucose uptake rate, insulin resistance

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57 The Healing 'Touch' of Music: A Neuro-Acoustics Approach to Understand Its Therapeutic Effect

Authors: Jagmeet S. Kanwal, Julia F. Langley

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Music can heal the body, but a mechanistic understanding of this phenomenon is lacking. This study explores the effects of music presentation on neurologic and physiologic responses leading to metabolic changes in the human body. The mind and body co-exist in a corporeal entity and within this framework, sickness ensues when the mind-body balance goes awry. It is further hypothesized that music has the capacity to directly reset this balance. Two lines of inquiry taken together can provide a mechanistic understanding of this phenomenon 1) Empirical evidence for a sound-sensitive pressure sensor system in the body, and 2) The notion of a “healing center” within the brain that is activated by specific patterns of sounds. From an acoustics perspective, music is spatially distributed as pressure waves ranging from a few cm to several meters in wavelength. These waves interact and propagate in three-dimensions in unique ways, depending on the wavelength. Furthermore, music creates dynamically changing wave-fronts. Frequencies between 200 Hz and 1 kHz generate wavelengths that range from 5'6" to 1 foot. These dimensions are in the range of the body size of most people making it plausible that these pressure waves can geometrically interact with the body surface and create distinct patterns of pressure stimulation across the skin surface. For humans, short wavelength, high frequency (> 200 Hz) sounds are best received via cochlear receptors. For low frequency (< 200 Hz), long wavelength sound vibrations, however, the whole body may act as an ideal receiver. A vast array of highly sensitive pressure receptors (Pacinian corpuscles) is present just beneath the skin surface, as well as in the tendons, bones, several organs in the abdomen, and the sexual organs. Per the available empirical evidence, these receptors contribute to music perception by allowing the whole body to function as a sound receiver, and knowledge of how they function is essential to fully understanding the therapeutic effect of music. Neuroscientific studies have established that music stimulates the limbic system that can trigger states of anxiety, arousal, fear, and other emotions. These emotional states of brain activity play a crucial role in filtering top-down feedback from thoughts and bottom-up sensory inputs to the autonomic system, which automatically regulates bodily functions. Music likely exerts its pleasurable and healing effects by enhancing functional and effective connectivity and feedback mechanisms between brain regions that mediate reward, autonomic, and cognitive processing. Stimulation of pressure receptors under the skin by low-frequency music-induced sensations can activate multiple centers in the brain, including the amygdala, the cingulate cortex, and nucleus accumbens. Melodies in music in the low (< 600 Hz) frequency range may augment auditory inputs after convergence of the pressure-sensitive inputs from the vagus nerve onto emotive processing regions within the limbic system. The integration of music-generated auditory and somato-visceral inputs may lead to a synergistic input to the brain that promotes healing. Thus, music can literally heal humans through “touch” as it energizes the brain’s autonomic system for restoring homeostasis.

Keywords: acoustics, brain, music healing, pressure receptors

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56 Metabolic Changes during Reprogramming of Wheat and Triticale Microspores

Authors: Natalia Hordynska, Magdalena Szechynska-Hebda, Miroslaw Sobczak, Elzbieta Rozanska, Joanna Troczynska, Zofia Banaszak, Maria Wedzony

Abstract:

Albinism is a common problem encountered in wheat and triticale breeding programs, which require in vitro culture steps e.g. generation of doubled haploids via androgenesis process. Genetic factor is a major determinant of albinism, however, environmental conditions such as temperature and media composition influence the frequency of albino plant formation. Cold incubation of wheat and triticale spikes induced a switch from gametophytic to sporophytic development. Further, androgenic structures formed from anthers of the genotypes susceptible to androgenesis or treated with cold stress, had a pool of structurally primitive plastids, with small starch granules or swollen thylakoids. High temperature was a factor inducing andro-genesis of wheat and triticale, but at the same time, it was a factor favoring the formation of albino plants. In genotypes susceptible to albinism or after heat stress conditions, cells formed from anthers were vacuolated, and plastids were eliminated. Partial or complete loss of chlorophyll pigments and incomplete differentiation of chloroplast membranes result in formation of tissues or whole plant unable to perform photosynthesis. Indeed, susceptibility to the andro-genesis process was associated with an increase of total concentration of photosynthetic pigments in anthers, spikes and regenerated plants. The proper balance of the synthesis of various pigments, was the starting point for their proper incorporation into photosynthetic membranes. In contrast, genotypes resistant to the androgenesis process and those treated with heat, contained 100 times lower content of photosynthetic pigments. In particular, the synthesis of violaxanthin, zeaxanthin, lutein and chlorophyll b was limited. Furthermore, deregulation of starch and lipids synthesis, which led to the formation of very complex starch granules and an increased number of oleosomes, respectively, correlated with the reduction of the efficiency of androgenesis. The content of other sugars varied depending on the genotype and the type of stress. The highest content of various sugars was found for genotypes susceptible to andro-genesis, and highly reduced for genotypes resistant to androgenesis. The most important sugars seem to be glucose and fructose. They are involved in sugar sensing and signaling pathways, which affect the expression of various genes and regulate plant development. Sucrose, on the other hand, seems to have minor effect at each stage of the androgenesis. The sugar metabolism was related to metabolic activity of microspores. The genotypes susceptible to androgenesis process had much faster mitochondrium- and chloroplast-dependent energy conversion and higher heat production by tissues. Thus, the effectiveness of metabolic processes, their balance and the flexibility under the stress was a factor determining the direction of microspore development, and in the later stages of the androgenesis process, a factor supporting the induction of androgenic structures, chloroplast formation and the regeneration of green plants. The work was financed by Ministry of Agriculture and Rural Development within Program: ‘Biological Progress in Plant Production’, project no HOR.hn.802.15.2018.

Keywords: androgenesis, chloroplast, metabolism, temperature stress

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55 Effects of Heart Rate Variability Biofeedback to Improve Autonomic Nerve Function, Inflammatory Response and Symptom Distress in Patients with Chronic Kidney Disease: A Randomized Control Trial

Authors: Chia-Pei Chen, Yu-Ju Chen, Yu-Juei Hsu

Abstract:

The prevalence and incidence of end-stage renal disease in Taiwan ranks the highest in the world. According to the statistical survey of the Ministry of Health and Welfare in 2019, kidney disease is the ninth leading cause of death in Taiwan. It leads to autonomic dysfunction, inflammatory response and symptom distress, and further increases the damage to the structure and function of the kidneys, leading to increased demand for renal replacement therapy and risks of cardiovascular disease, which also has medical costs for the society. If we can intervene in a feasible manual to effectively regulate the autonomic nerve function of CKD patients, reduce the inflammatory response and symptom distress. To prolong the progression of the disease, it will be the main goal of caring for CKD patients. This study aims to test the effect of heart rate variability biofeedback (HRVBF) on improving autonomic nerve function (Heart Rate Variability, HRV), inflammatory response (Interleukin-6 [IL-6], C reaction protein [CRP] ), symptom distress (Piper fatigue scale, Pittsburgh Sleep Quality Index [PSQI], and Beck Depression Inventory-II [BDI-II] ) in patients with chronic kidney disease. This study was experimental research, with a convenience sampling. Participants were recruited from the nephrology clinic at a medical center in northern Taiwan. With signed informed consent, participants were randomly assigned to the HRVBF or control group by using the Excel BINOMDIST function. The HRVBF group received four weekly hospital-based HRVBF training, and 8 weeks of home-based self-practice was done with StressEraser. The control group received usual care. We followed all participants for 3 months, in which we repeatedly measured their autonomic nerve function (HRV), inflammatory response (IL-6, CRP), and symptom distress (Piper fatigue scale, PSQI, and BDI-II) on their first day of study participation (baselines), 1 month, and 3 months after the intervention to test the effects of HRVBF. The results were analyzed by SPSS version 23.0 statistical software. The data of demographics, HRV, IL-6, CRP, Piper fatigue scale, PSQI, and BDI-II were analyzed by descriptive statistics. To test for differences between and within groups in all outcome variables, it was used by paired sample t-test, independent sample t-test, Wilcoxon Signed-Rank test and Mann-Whitney U test. Results: Thirty-four patients with chronic kidney disease were enrolled, but three of them were lost to follow-up. The remaining 31 patients completed the study, including 15 in the HRVBF group and 16 in the control group. The characteristics of the two groups were not significantly different. The four-week hospital-based HRVBF training combined with eight-week home-based self-practice can effectively enhance the parasympathetic nerve performance for patients with chronic kidney disease, which may against the disease-related parasympathetic nerve inhibition. In the inflammatory response, IL-6 and CRP in the HRVBF group could not achieve significant improvement when compared with the control group. Self-reported fatigue and depression significantly decreased in the HRVBF group, but they still failed to achieve a significant difference between the two groups. HRVBF has no significant effect on improving the sleep quality for CKD patients.

Keywords: heart rate variability biofeedback, autonomic nerve function, inflammatory response, symptom distress, chronic kidney disease

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54 Anesthesia for Spinal Stabilization Using Neuromuscular Blocking Agents in Dog: Case Report

Authors: Agata Migdalska, Joanna Berczynska, Ewa Bieniek, Jacek Sterna

Abstract:

Muscle relaxation is considered important during general anesthesia for spine stabilization. In a presented case peripherally acting muscle relaxant was applied during general anesthesia for spine stabilization surgery. The patient was a dog, 11-years old, 26 kg, male, mix breed. Spine fracture was situated between Th13-L1-L2, probably due to the car accident. Preanesthetic physical examination revealed no sign underlying health issues. The dog was premedicated with midazolam 0.2 mg IM and butorphanol 2.4 mg IM. General anesthesia was induced with propofol IV. After the induction, the dog was intubated with an endotracheal tube and connected to an open-ended rebreathing system and maintained with the use of inhalation anesthesia with isoflurane in oxygen. 0,5 mg/ kg of rocuronium was given IV. Use of muscle relaxant was accompanied by an assessment of the degree of neuromuscular blockade by peripheral nerve stimulator. Electrodes were attached to the skin overlying at the peroneal nerve at the lateral cranial tibia. Four electrical pulses were applied to the nerve over a 2 second period. When satisfying nerve block was detected dog was prepared for the surgery. No further monitoring of the effectiveness of blockade was performed during surgery. Mechanical ventilation was kept during anesthesia. During surgery dog maintain stable, and no anesthesiological complication occur. Intraoperatively surgeon claimed that neuromuscular blockade results in a better approach to the spine and easier muscle manipulation which was helpful in order to see the fracture and replace bone fragments. Finally, euthanasia was performed intraoperatively as a result of vast myelomalacia process of the spinal cord. This prevented examination of the recovering process. Neuromuscular blocking agents act at the neuromuscular junction to provide profound muscle relaxation throughout the body. Muscle blocking agents are neither anesthetic nor analgesic; therefore inappropriately used may cause paralysis in fully conscious and feeling pain patient. They cause paralysis of all skeletal muscles, also diaphragm and intercostal muscles when given in higher doses. Intraoperative management includes maintaining stable physiological conditions, which involves adjusting hemodynamic parameters, ensuring proper ventilation, avoiding variations in temperature, maintain normal blood flow to promote proper oxygen exchange. Neuromuscular blocking agent can cause many side effects like residual paralysis, anaphylactic or anaphylactoid reactions, delayed recovery from anesthesia, histamine release, recurarization. Therefore reverse drug like neostigmine (with glikopyrolat) or edrofonium (with atropine) should be used in case of a life-threatening situation. Another useful drug is sugammadex, although the cost of this drug strongly limits its use. Muscle relaxant improves surgical conditions during spinal surgery, especially in heavily muscled individuals. They are also used to facilitate the replacement of dislocated joints as they improve conditions during fracture reduction. It is important to emphasize that in a patient with muscle weakness neuromuscular blocking agents may result in intraoperative and early postoperative cardiovascular and respiratory complications, as well as prolonged recovery from anesthesia. This should not appear in patients with recent spine fracture or luxation. Therefore it is believed that neuromuscular blockers could be useful during spine stabilization procedures.

Keywords: anesthesia, dog, neuromuscular block, spine surgery

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53 User-Controlled Color-Changing Textiles: From Prototype to Mass Production

Authors: Joshua Kaufman, Felix Tan, Morgan Monroe, Ayman Abouraddy

Abstract:

Textiles and clothing have been a staple of human existence for millennia, yet the basic structure and functionality of textile fibers and yarns has remained unchanged. While color and appearance are essential characteristics of a textile, an advancement in the fabrication of yarns that allows for user-controlled dynamic changes to the color or appearance of a garment has been lacking. Touch-activated and photosensitive pigments have been used in textiles, but these technologies are passive and cannot be controlled by the user. The technology described here allows the owner to control both when and in what pattern the fabric color-change takes place. In addition, the manufacturing process is compatible with mass-producing the user-controlled, color-changing yarns. The yarn fabrication utilizes a fiber spinning system that can produce either monofilament or multifilament yarns. For products requiring a more robust fabric (backpacks, purses, upholstery, etc.), larger-diameter monofilament yarns with a coarser weave are suitable. Such yarns are produced using a thread-coater attachment to encapsulate a 38-40 AWG metal wire inside a polymer sheath impregnated with thermochromic pigment. Conversely, products such as shirts and pants requiring yarns that are more flexible and soft against the skin comprise multifilament yarns of much smaller-diameter individual fibers. Embedding a metal wire in a multifilament fiber spinning process has not been realized to date. This research has required collaboration with Hills, Inc., to design a liquid metal-injection system to be combined with fiber spinning. The new system injects molten tin into each of 19 filaments being spun simultaneously into a single yarn. The resulting yarn contains 19 filaments, each with a tin core surrounded by a polymer sheath impregnated with thermochromic pigment. The color change we demonstrate is distinct from garments containing LEDs that emit light in various colors. The pigment itself changes its optical absorption spectrum to appear a different color. The thermochromic color-change is induced by a temperature change in the inner metal wire within each filament when current is applied from a small battery pack. The temperature necessary to induce the color change is near body temperature and not noticeable by touch. The prototypes already developed either use a simple push button to activate the battery pack or are wirelessly activated via a smart-phone app over Wi-Fi. The app allows the user to choose from different activation patterns of stripes that appear in the fabric continuously. The power requirements are mitigated by a large hysteresis in the activation temperature of the pigment and the temperature at which there is full color return. This was made possible by a collaboration with Chameleon International to develop a new, customized pigment. This technology enables a never-before seen capability: user-controlled, dynamic color and pattern change in large-area woven and sewn textiles and fabrics with wide-ranging applications from clothing and accessories to furniture and fixed-installation housing and business décor. The ability to activate through Wi-Fi opens up possibilities for the textiles to be part of the ‘Internet of Things.’ Furthermore, this technology is scalable to mass-production levels for wide-scale market adoption.

Keywords: activation, appearance, color, manufacturing

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52 Impact of Ocean Acidification on Gene Expression Dynamics during Development of the Sea Urchin Species Heliocidaris erythrogramma

Authors: Hannah R. Devens, Phillip L. Davidson, Dione Deaker, Kathryn E. Smith, Gregory A. Wray, Maria Byrne

Abstract:

Marine invertebrate species with calcifying larvae are especially vulnerable to ocean acidification (OA) caused by rising atmospheric CO₂ levels. Acidic conditions can delay development, suppress metabolism, and decrease the availability of carbonate ions in the ocean environment for skeletogenesis. These stresses often result in increased larval mortality, which may lead to significant ecological consequences including alterations to the larval settlement, population distribution, and genetic connectivity. Importantly, many of these physiological and developmental effects are caused by genetic and molecular level changes. Although many studies have examined the effect of near-future oceanic pH levels on gene expression in marine invertebrates, little is known about the impact of OA on gene expression in a developmental context. Here, we performed mRNA-sequencing to investigate the impact of environmental acidity on gene expression across three developmental stages in the sea urchin Heliocidaris erythrogramma. We collected RNA from gastrula, early larva, and 1-day post-metamorphic juvenile sea urchins cultured at present-day and predicted future oceanic pH levels (pH 8.1 and 7.7, respectively). We assembled an annotated reference transcriptome encompassing development from egg to ten days post-metamorphosis by combining these data with datasets from two previous developmental transcriptomic studies of H. erythrogramma. Differential gene expression and time course analyses between pH conditions revealed significant alterations to developmental transcription that are potentially associated with pH stress. Consistent with previous investigations, genes involved in biomineralization and ion transport were significantly upregulated under acidic conditions. Differences in gene expression between the two pH conditions became more pronounced post-metamorphosis, suggesting a development-dependent effect of OA on gene expression. Furthermore, many differences in gene expression later in development appeared to be a result of broad downregulation at pH 7.7: of 539 genes differentially expressed at the juvenile stage, 519 of these were lower in the acidic condition. Time course comparisons between pH 8.1 and 7.7 samples also demonstrated over 500 genes were more lowly expressed in pH 7.7 samples throughout development. Of the genes exhibiting stage-dependent expression level changes, over 15% of these diverged from the expected temporal pattern of expression in the acidic condition. Through these analyses, we identify novel candidate genes involved in development, metabolism, and transcriptional regulation that are possibly affected by pH stress. Our results demonstrate that pH stress significantly alters gene expression dynamics throughout development. A large number of genes differentially expressed between pH conditions in juveniles relative to earlier stages may be attributed to the effects of acidity on transcriptional regulation, as a greater proportion of mRNA at this later stage has been nascent transcribed rather than maternally loaded. Also, the overall downregulation of many genes in the acidic condition suggests that OA-induced developmental delay manifests as suppressed mRNA expression, possibly from lower transcription rates or increased mRNA degradation in the acidic environment. Further studies will be necessary to determine in greater detail the extent of OA effects on early developing marine invertebrates.

Keywords: development, gene expression, ocean acidification, RNA-sequencing, sea urchins

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51 Intelligent Materials and Functional Aspects of Shape Memory Alloys

Authors: Osman Adiguzel

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Shape-memory alloys are a new class of functional materials with a peculiar property known as shape memory effect. These alloys return to a previously defined shape on heating after deformation in low temperature product phase region and take place in a class of functional materials due to this property. The origin of this phenomenon lies in the fact that the material changes its internal crystalline structure with changing temperature. Shape memory effect is based on martensitic transitions, which govern the remarkable changes in internal crystalline structure of materials. Martensitic transformation, which is a solid state phase transformation, occurs in thermal manner in material on cooling from high temperature parent phase region. This transformation is governed by changes in the crystalline structure of the material. Shape memory alloys cycle between original and deformed shapes in bulk level on heating and cooling, and can be used as a thermal actuator or temperature-sensitive elements due to this property. Martensitic transformations usually occur with the cooperative movement of atoms by means of lattice invariant shears. The ordered parent phase structures turn into twinned structures with this movement in crystallographic manner in thermal induced case. The twinned martensites turn into the twinned or oriented martensite by stressing the material at low temperature martensitic phase condition. The detwinned martensite turns into the parent phase structure on first heating, first cycle, and parent phase structures turn into the twinned and detwinned structures respectively in irreversible and reversible memory cases. On the other hand, shape memory materials are very important and useful in many interdisciplinary fields such as medicine, pharmacy, bioengineering, metallurgy and many engineering fields. The choice of material as well as actuator and sensor to combine it with the host structure is very essential to develop main materials and structures. Copper based alloys exhibit this property in metastable beta-phase region, which has bcc-based structures at high temperature parent phase field, and these structures martensitically turn into layered complex structures with lattice twinning following two ordered reactions on cooling. Martensitic transition occurs as self-accommodated martensite with inhomogeneous shears, lattice invariant shears which occur in two opposite directions, <110 > -type directions on the {110}-type plane of austenite matrix which is basal plane of martensite. This kind of shear can be called as {110}<110> -type mode and gives rise to the formation of layered structures, like 3R, 9R or 18R depending on the stacking sequences on the close-packed planes of the ordered lattice. In the present contribution, x-ray diffraction and transmission electron microscopy (TEM) studies were carried out on two copper based alloys which have the chemical compositions in weight; Cu-26.1%Zn 4%Al and Cu-11%Al-6%Mn. X-ray diffraction profiles and electron diffraction patterns reveal that both alloys exhibit super lattice reflections inherited from parent phase due to the displacive character of martensitic transformation. X-ray diffractograms taken in a long time interval show that locations and intensities of diffraction peaks change with the aging time at room temperature. In particular, some of the successive peak pairs providing a special relation between Miller indices come close each other.

Keywords: Shape memory effect, martensite, twinning, detwinning, self-accommodation, layered structures

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50 Complex Dynamics in a Morphologically Heterogeneous Biological Medium

Authors: Turky Al-Qahtani, Roustem Miftahof

Abstract:

Introduction: Under common assumptions of excitabi-lity, morphological (cellular) homogeneity, and spatial structural anomalies added as required, it has been shown that biological systems are able to display travelling wave dynamics. Being not self-sustainable, existence depends on the electrophysiological state of transmembrane ion channels and it requires an extrinsic/intrinsic periodic source. However, organs in the body are highly multicellular, heterogeneous, and their functionality is the outcome of electro-mechanical conjugation, rather than excitability only. Thus, peristalsis in the gut relies on spatiotemporal myoelectrical pattern formations between the mechanical, represented by smooth muscle cells (SM), and the control, comprised of a chain of primary sensory and motor neurones, components. Synaptically linked through the afferent and efferent pathways, they form a functional unit (FU) of the gut. Aims: These are: i) to study numerically the complex dynamics, and ii) to investigate the possibility of self-sustained myoelectrical activity in the FU. Methods: The FU recreates the following sequence of physiological events: deformation of mechanoreceptors of located in SM; generation and propagation of electrical waves of depolarisation - spikes - along the axon to the soma of the primary neurone; discharge of the primary neurone and spike propagation towards the motor neurone; burst of the motor neurone and transduction of spikes to SM, subsequently producing forces of contraction. These are governed by a system of nonlinear partial and ordinary differential equations being a modified version of the Hodgkin-Huxley model and SM fibre mechanics. In numerical experiments; the source of excitation is mechanical stretches of SM at a fixed amplitude and variable frequencies. Results: Low frequency (0.5 < v < 2 Hz) stimuli cause the propagation of spikes in the neuronal chain and, finally, the generation of active forces by SM. However, induced contractions are not sufficient to initiate travelling wave dynamics in the control system. At frequencies, 2 < v < 4 Hz, multiple low amplitude and short-lasting contractions are observed in SM after the termination of stretching. For frequencies (0.5 < v < 4 Hz), primary and sensory neurones demonstrate strong connectivity and coherent electrical activity. Significant qualitative and quantitative changes in dynamics of myoelectical patterns with a transition to a self-organised mode are recorded with the high degree of stretches at v = 4.5 Hz. Increased rates of deformation lead to the production of high amplitude signals at the mechanoreceptors with subsequent self-sustained excitation within the neuronal chain. Remarkably, the connection between neurones weakens resulting in incoherent firing. Further increase in a frequency of stimulation (v > 4.5 Hz) has a detrimental effect on the system. The mechanical and control systems become disconnected and exhibit uncoordinated electromechanical activity. Conclusion: To our knowledge, the existence of periodic activity in a multicellular, functionally heterogeneous biological system with mechano-electrical dynamics, such as the FU, has been demonstrated for the first time. These findings support the notion of possible peristalsis in the gut even in the absence of intrinsic sources - pacemaker cells. Results could be implicated in the pathogenesis of intestinal dysrythmia, a medical condition associated with motor dysfunction.

Keywords: complex dynamics, functional unit, the gut, dysrythmia

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49 Reimagining Landscapes: Psychological Responses and Behavioral Shifts in the Aftermath of the Lytton Creek Fire

Authors: Tugba Altin

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In an era where the impacts of climate change resonate more pronouncedly than ever, communities globally grapple with events bearing both tangible and intangible ramifications. Situating this within the evolving landscapes of Psychological and Behavioral Sciences, this research probes the profound psychological and behavioral responses evoked by such events. The Lytton Creek Fire of 2021 epitomizes these challenges. While tangible destruction is immediate and evident, the intangible repercussions—emotional distress, disintegration of cultural landscapes, and disruptions in place attachment (PA)—require meticulous exploration. PA, emblematic of the emotional and cognitive affiliations individuals nurture with their environments, emerges as a cornerstone for comprehending how environmental cataclysms influence cultural identity and bonds to land. This study, harmonizing the core tenets of an interpretive phenomenological approach with a hermeneutic framework, underscores the pivotal nature of this attachment. It delves deep into the realm of individuals' experiences post the Lytton Creek Fire, unraveling the intricate dynamics of PA amidst such calamity. The study's methodology deviates from conventional paradigms. Instead of traditional interview techniques, it employs walking audio sessions and photo elicitation methods, granting participants the agency to immerse, re-experience, and vocalize their sentiments in real-time. Such techniques shed light on spatial narratives post-trauma and capture the otherwise elusive emotional nuances, offering a visually rich representation of place-based experiences. Central to this research is the voice of the affected populace, whose lived experiences and testimonies form the nucleus of the inquiry. As they renegotiate their bonds with transformed environments, their narratives reveal the indispensable role of cultural landscapes in forging place-based identities. Such revelations accentuate the necessity of integrating both tangible and intangible trauma facets into community recovery strategies, ensuring they resonate more profoundly with affected individuals. Bridging the domains of environmental psychology and behavioral sciences, this research accentuates the intertwined nature of tangible restoration with the imperative of emotional and cultural recuperation post-environmental disasters. It advocates for adaptation initiatives that are rooted in the lived realities of the affected, emphasizing a holistic approach that recognizes the profundity of human connections to landscapes. This research advocates the interdisciplinary exchange of ideas and strategies in addressing post-disaster community recovery strategies. It not only enriches the climate change discourse by emphasizing the human facets of disasters but also reiterates the significance of an interdisciplinary approach, encompassing psychological and behavioral nuances, for fostering a comprehensive understanding of climate-induced traumas. Such a perspective is indispensable for shaping more informed, empathetic, and effective adaptation strategies.

Keywords: place attachment, community recovery, disaster response, restorative landscapes, sensory response, visual methodologies

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48 Development of Peptide Inhibitors against Dengue Virus Infection by in Silico Design

Authors: Aussara Panya, Nunghathai Sawasdee, Mutita Junking, Chatchawan Srisawat, Kiattawee Choowongkomon, Pa-Thai Yenchitsomanus

Abstract:

Dengue virus (DENV) infection is a global public health problem with approximately 100 million infected cases a year. Presently, there is no approved vaccine or effective drug available; therefore, the development of anti-DENV drug is urgently needed. The clinical reports revealing the positive association between the disease severity and viral titer has been reported previously suggesting that the anti-DENV drug therapy can possibly ameliorate the disease severity. Although several anti-DENV agents showed inhibitory activities against DENV infection, to date none of them accomplishes clinical use in the patients. The surface envelope (E) protein of DENV is critical for the viral entry step, which includes attachment and membrane fusion; thus, the blocking of envelope protein is an attractive strategy for anti-DENV drug development. To search the safe anti-DENV agent, this study aimed to search for novel peptide inhibitors to counter DENV infection through the targeting of E protein using a structure-based in silico design. Two selected strategies has been used including to identify the peptide inhibitor which interfere the membrane fusion process whereby the hydrophobic pocket on the E protein was the target, the destabilization of virion structure organization through the disruption of the interaction between the envelope and membrane proteins, respectively. The molecular docking technique has been used in the first strategy to search for the peptide inhibitors that specifically bind to the hydrophobic pocket. The second strategy, the peptide inhibitor has been designed to mimic the ectodomain portion of membrane protein to disrupt the protein-protein interaction. The designed peptides were tested for the effects on cell viability to measure the toxic to peptide to the cells and their inhibitory assay to inhibit the DENV infection in Vero cells. Furthermore, their antiviral effects on viral replication, intracellular protein level and viral production have been observed by using the qPCR, cell-based flavivirus immunodetection and immunofluorescence assay. None of tested peptides showed the significant effect on cell viability. The small peptide inhibitors achieved from molecular docking, Glu-Phe (EF), effectively inhibited DENV infection in cell culture system. Its most potential effect was observed for DENV2 with a half maximal inhibition concentration (IC50) of 96 μM, but it partially inhibited other serotypes. Treatment of EF at 200 µM on infected cells also significantly reduced the viral genome and protein to 83.47% and 84.15%, respectively, corresponding to the reduction of infected cell numbers. An additional approach was carried out by using peptide mimicking membrane (M) protein, namely MLH40. Treatment of MLH40 caused the reduction of foci formation in four individual DENV serotype (DENV1-4) with IC50 of 24-31 μM. Further characterization suggested that the MLH40 specifically blocked viral attachment to host membrane, and treatment with 100 μM could diminish 80% of viral attachment. In summary, targeting the hydrophobic pocket and M-binding site on the E protein by using the peptide inhibitors could inhibit DENV infection. The results provide proof of-concept for the development of antiviral therapeutic peptide inhibitors to counter DENV infection through the use of a structure-based design targeting conserved viral protein.

Keywords: dengue virus, dengue virus infection, drug design, peptide inhibitor

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