Search results for: nerve differentiation

Authors: Gehan ElAkabawy

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Background: Diabetes mellitus causes severe deteriorations of almost all the organs and systems of the body, as well as significant damage to the oral cavity. The oral changes are mainly related to salivary glands dysfunction characterized by hyposalivation and xerostomia, which significantly reduce diabetic patients’ quality of life. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that the trophic support mediated by dental pulp stem cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of human dental pulp stem cells in diabetic-induced parotid gland damage have not been previously investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of human dental pulp stem cells (hDPSCs) on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. Methods: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ+hDPSCs). hDPSCs or vehicle was injected into the tail vein 7 days after STZ injection. The fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA) expression in parotid tissues were assessed 28 days post-transplantation. Results: Transplantation of hDPSCs downregulated blood glucose, improved the salivary flow rate, and reduced oxidative stress. The cells migrated to, survived, and differentiated into acinar, ductal, and myoepithelial cells in the STZ-injured parotid gland. Moreover, they downregulated the expression of caspase-3 and upregulated the expression of VEGF and PCNA, likely exerting pro-angiogenetic and antiapoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide (NO) -tetrahydrobiopterin (BH4) pathway. Conclusions: Our results show that hDPSCs can migrate to and survive within the STZ-injured parotid gland, where they prevent its functional and morphological damage by restoring normal glucose levels, differentiating into parotid cell populations, and stimulating paracrine-mediated regeneration. Thus, hDPSCs may have therapeutic potential in the treatment of diabetes-induced parotid gland injury.

Keywords: dental pulp stem cells, diabetes, streptozotocin, parotid gland

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Authors: Kim Ji-Hyun, Jeon Hye-Seon, Kwon Oh-Yun, Park Eun-Young, Hwang Ui-Jae, Gwak Gyeong-Tae, Yoon Hyeo-Bin

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Stress urinary incontinence (SUI), a common women health problem, is an involuntary leakage of urine while sneezing, coughing, or physical exertion caused by insufficient strength of the pelvic floor and sphincter muscles. SUI also leads to decrease in quality of life and limits sexual activities. SUI is related to the increased bladder neck angle, bladder neck movement, funneling index, urethral width, and decreased urethral length. Various pelvic floor muscle electrical stimulation (ES) interventions have been applied to improve the symptoms of the people with SUI. ES activates afferent fibers of pudendal nerve and smoothly induces contractions of the pelvic floor muscles such as striated periurethral muscles and striated pelvic floor muscles. ES via intravaginal electrodes are the most frequently used types of the pelvic floor muscle ES for the female SUI. However, inserted electrode is uncomfortable and it increases the risks of infection. The purpose of this preliminary study was to determine if the 8-week transcutaneous pelvic floor ES would be effective to improve the symptoms and satisfaction of the females with SUI. Easy-K, specially designed ES equipment for the people with SUI, was used in this study. The oval shape stimulator can be placed on a toilet seat, and the surface has invaded electrode fit to contact with the entire vulva area while users are sitting on the stimulator. Five women with SUI were included in this experiment. Prior to the participation, subjects were instructed about procedures and precautions in using the ES. They have used the stimulator once a day for 20 minutes for each session at home. Outcome data was collected 3 times at the baseline, 4 weeks and 8 weeks after the intervention. Intravaginal sonography was used to measure the bladder neck angle, bladder neck movement, funneling index, thickness of an anterior rhabdosphincter and a posterior rhabdosphincter, urethral length, and urethral width. Leavator ani muscle (LAM) contraction strength was assessed by manual palpation according to the oxford scoring system. In addition, incontinence quality of life (IQOL) and female sexual function index (FSFI) questionnaires were used to obtain addition subjective information. Friedman test, a nonparametric statistical test, was used to determine the effectiveness of the ES. The Wilcoxon test was used for the post-hoc analysis and the significance level was set at .05. The bladder neck angle, funneling index and urethral width were significantly decreased after 8-weeks of intervention (p<.05). LAM contraction score, urethral length and anterior and posterior rhabdosphicter thickness were statistically increased by the intervention (p<.05). However, no significant change was found in the bladder neck movement. Although total score of the IQOL did not improve, the score of the ‘avoidance’ subscale of IQOL had significant improved (p<.05). FSFI had statistical difference in FSFI total score and ‘desire’ subscale (p<.05). In conclusion, 8-week use of a transcutaneous ES on peri-vulva area improved dynamic mechanical structures of the pelvic floor musculature as well as IQOL and conjugal relationship.

Keywords: electrical stimulation, Pelvic floor muscle, sonography, stress urinary incontinence, women health

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Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

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Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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Authors: Yuanyuan Fu

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Background and Purpose: Along with the change of society and economy, the traditional home function of old people has gradually weakened in the contemporary China. Acknowledging these situations, to better meet old people’s needs on formal services and improve the quality of later life, this study seeks to identify patterns of intended service use among frail old people living in the communities and examined determinants that explain heterogeneous variations in old people’s intended service use patterns. Additionally, this study also tested the relationship between culture value and intended service use patterns and the mediating role of enabling factors in terms of culture value and intended service use patterns. Methods：Participants were recruited from Haidian District, Beijing, China in 2015. The multi-stage sampling method was adopted to select sub-districts, communities and old people aged 70 years old or older. After screening, 577 old people with limitations in daily life, were successfully interviewed. After data cleaning, 550 samples were included for data analysis. This study establishes a conceptual framework based on the Anderson Model (including predisposing factors, enabling factors and need factors), and further developed it by adding culture value factors (including attitudes towards filial piety and attitudes towards social face). Using a latent class analysis (LCA), this study classifies overall patterns of old people’s formal service utilization. Fourteen types of formal services were taken into account, including housework, voluntary support, transportation, home-delivered meals, and home-delivery medical care, elderly’s canteen and day-care center/respite care and so on. Structural equation modeling (SEM) was used to examine the direct effect of culture value on service use pattern, and the mediating effect of the enabling factors. Results: The LCA classified a hierarchical structure of service use patterns: multiple intended service use (N=69, 23%), selective intended service use (N=129, 23%), and light intended service use (N=352, 64%). Through SEM, after controlling predisposing factors and need factors, the results showed the significant direct effect of culture value on older people’s intended service use patterns. Enabling factors had a partial mediation effect on the relationship between culture value and the patterns. Conclusions and Implications: Differentiation of formal services may be important for meeting frail old people’s service needs and distributing program resources by identifying target populations for intervention, which may make reference to specific interventions to better support frail old people. Additionally, culture value had a unique direct effect on the intended service use patterns of frail old people in China, enriching our theoretical understanding of sources of culture value and their impacts. The findings also highlighted the mediation effects of enabling factors on the relationship between culture value factors and intended service use patterns. This study suggests that researchers and service providers should pay more attention to the important role of culture value factors in contributing to intended service use patterns and also be more sensitive to the mediating effect of enabling factors when discussing the relationship between culture value and the patterns.

Keywords: frail old people, intended service use pattern, culture value, enabling factors, contemporary China, latent class analysis

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Authors: M. Cardenas-Garcia, P. P. Gonzalez-Perez

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Intercellular communication is a necessary condition for cellular functions and it allows a group of cells to survive as a population. Throughout this interaction, the cells work in a coordinated and collaborative way which facilitates their survival. In the case of cancerous cells, these take advantage of intercellular communication to preserve their malignancy, since through these physical unions they can send signs of malignancy. The Wnt/β-catenin signaling pathway plays an important role in the formation of intercellular communications, being also involved in a large number of cellular processes such as proliferation, differentiation, adhesion, cell survival, and cell death. The modeling and simulation of cellular signaling systems have found valuable support in a wide range of modeling approaches, which cover a wide spectrum ranging from mathematical models; e.g., ordinary differential equations, statistical methods, and numerical methods– to computational models; e.g., process algebra for modeling behavior and variation in molecular systems. Based on these models, different simulation tools have been developed from mathematical ones to computational ones. Regarding cellular and molecular processes in cancer, its study has also found a valuable support in different simulation tools that, covering a spectrum as mentioned above, have allowed the in silico experimentation of this phenomenon at the cellular and molecular level. In this work, we simulate and explore the complex interaction patterns of intercellular communication in cancer cells using the Cellulat bioinformatics tool, a computational simulation tool developed by us and motivated by two key elements: 1) a biochemically inspired model of self-organizing coordination in tuple spaces, and 2) the Gillespie’s algorithm, a stochastic simulation algorithm typically used to mimic systems of chemical/biochemical reactions in an efficient and accurate way. The main idea behind the Cellulat simulation tool is to provide an in silico experimentation environment that complements and guides in vitro experimentation in intra and intercellular signaling networks. Unlike most of the cell signaling simulation tools, such as E-Cell, BetaWB and Cell Illustrator which provides abstractions to model only intracellular behavior, Cellulat is appropriate for modeling both intracellular signaling and intercellular communication, providing the abstractions required to model –and as a result, simulate– the interaction mechanisms that involve two or more cells, that is essential in the scenario discussed in this work. During the development of this work we made evident the application of our computational simulation tool (Cellulat) for the modeling and simulation of intercellular communication between normal and cancerous cells, and in this way, propose key molecules that may prevent the arrival of malignant signals to the cells that surround the tumor cells. In this manner, we could identify the significant role that has the Wnt/β-catenin signaling pathway in cellular communication, and therefore, in the dissemination of cancer cells. We verified, using in silico experiments, how the inhibition of this signaling pathway prevents that the cells that surround a cancerous cell are transformed.

Keywords: cancer cells, in silico approach, intercellular communication, key molecules, modeling and simulation

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Authors: Mustafa Cam, Nedim Ongun, Ufuk Kutluana

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Introduction: Restless LegsSyndrome (RLS) is a common, under-recognized disorder disrupts sleep and diminishes quality of life (1). The most common conditions highly associated with RLS include renalfailure, iron and folic acid deficiency, peripheral neuropathy, pregnancy, celiacdisease, Crohn’sdiseaseandrarelymalignancy (2).Despite a clear relation between low peripheral iron and increased prevalence and severity of RLS, the prevalence and clinical significance of RLS in iron-deficientanemic populations is unknown (2). We report here a case of RLS due to iron deficiency in the setting of neuroendocrinetumor. Report of Case: A 35 year-old man was referred to our clinic with general weakness, weight loss (10 kg in 2 months)and 2-month history of uncomfortable sensations in his legs with urge to move, partially relieved by movement. The symptoms were presented very day, worsening in the evening; the discomfort forced the patient to getup and walk around at night. RLS was severe, with a score of 22 at the International RLS ratingscale. The patient had no past medical history. The patient underwent a complete set of blood analyses and the following ab normal values were found (normal limitswithinbrackets): hemoglobin 9.9 g/dl (14-18), MCV 70 fL (80-94), ferritin 3,5 ng/mL (13-150). Brain and spinemagnetic resonance imaging was normal. The patient consultated with gastroenterology clinic and gastointestinal systemendoscopy was performed for theetiology of the iron deficiency anemia. After the gastricbiopsy, results allowed us to reach the diagnosis of neuroen docrine tumor and the patient referred to oncology clinic. Discussion: The first important consideration from this case report is that the patient was referred to our clinic because of his severe RLS symptoms dramatically reducing his quality of life. However, our clinical study clearly demonstrated that RLS was not the primary disease. Considering the information available for this patient, we believe that the most likely possibility is that RLS was secondary to iron deficiency, a very well-known and established cause of RLS in theliterature (3,4). Neuroendocrine tumors (NETs) are rare epithelial neoplasms with neuroendocrine differentiation that most commonly originate in the lungs and gastrointestinal tract (5). NETs vary widely in their clinical presentation; symptoms are often nonspecific and can be mistaken for those of other more common conditions (6). 50% of patients with reported disease stage have either regional or distant metastases at diagnosis (7). Accurate and earlier NET diagnosis is the first step in shortening the time to optimal care and improved outcomes for patients (8). The most important message from this case report is that RLS symptoms can sometimes be thesign of a life-threatening condition. Conclusion: Careful and complete collection of clinical and laboratory data should be carried out in RLS patients. Inparticular, if RLS onset coincides with weight loss and iron deficieny anemia, gastricendos copy should be performed. It is known about that malignancy is a rare etiology in RLS patients and to our knowledge; it is the first case with neuro endocrine tumor presenting with RLS.

Keywords: neurology, neuroendocrine tumor, restless legs syndrome, sleep

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Authors: Alexandra Sargent, Sarah Ferris, Ioannis Theofanous

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The Abbott RealTime High Risk HPV test (RealTime HPV) is one of five assays clinically validated and approved by the English NHS Cervical Screening Programme (CSP) for HPV triage of low grade dyskaryosis and test-of-cure of treated Cervical Intraepithelial Neoplasia. The assay is a highly automated multiplex real-time PCR test for detecting 14 high risk (hr) HPV types, with simultaneous differentiation of HPV 16 and HPV 18 versus non-HPV 16/18 hrHPV. An endogenous internal control ensures sample cellularity, controls extraction efficiency and PCR inhibition. The original cervical specimen collected in SurePath (SP) liquid-based cytology (LBC) medium (BD Diagnostics) and the SP post-gradient cell pellets (SPG) after cytological processing are both CE marked for testing with the RealTime HPV test. During the 2011 NHSCSP validation of new tests only the original aliquot of SP LBC medium was investigated. Residual sample volume left after cytology slide preparation is low and may not always have sufficient volume for repeat HPV testing or for testing of other biomarkers that may be implemented in testing algorithms in the future. The SPG samples, however, have sufficient volumes to carry out additional testing and necessary laboratory validation procedures. This study investigates the correlation of RealTime HPV results of cervical specimens collected in SP LBC medium from women with low grade cytological abnormalities observed with matched pairs of original SP LBC medium and SP post-gradient cell pellets (SPG) after cytology processing. Matched pairs of SP and SPG samples from 750 women with borderline (N = 392) and mild (N = 351) cytology were available for this study. Both specimen types were processed and parallel tested for the presence of hrHPV with RealTime HPV according to the manufacturer´s instructions. HrHPV detection rates and concordance between test results from matched SP and SPGCP pairs were calculated. A total of 743 matched pairs with valid test results on both sample types were available for analysis. An overall-agreement of hrHPV test results of 97.5% (k: 0.95) was found with matched SP/SPG pairs and slightly lower concordance (96.9%; k: 0.94) was observed on 392 pairs from women with borderline cytology compared to 351 pairs from women with mild cytology (98.0%; k: 0.95). Partial typing results were highly concordant in matched SP/SPG pairs for HPV 16 (99.1%), HPV 18 (99.7%) and non-HPV16/18 hrHPV (97.0%), respectively. 19 matched pairs were found with discrepant results: 9 from women with borderline cytology and 4 from women with mild cytology were negative on SPG and positive on SP; 3 from women with borderline cytology and 3 from women with mild cytology were negative on SP and positive on SPG. Excellent correlation of hrHPV DNA test results was found between matched pairs of SP original fluid and post-gradient cell pellets from women with low grade cytological abnormalities tested with the Abbott RealTime High-Risk HPV assay, demonstrating robust performance of the test with both specimen types and reassuring the utility of the assay for cytology triage with both specimen types.

Keywords: Abbott realtime test, HPV, SurePath liquid based cytology, surepath post-gradient cell pellet

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Authors: A. Chatel, I. S. Torreguitart, T. Verstraete

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The paper deals with a single point optimization of the LS89 turbine using an adjoint optimization and defining the design variables within a CAD system. The advantage of including the CAD model in the design system is that higher level constraints can be imposed on the shape, allowing the optimized model or component to be manufactured. However, CAD-based approaches restrict the design space compared to node-based approaches where every node is free to move. In order to preserve a rich design space, we develop a methodology to refine the CAD model during the optimization and to create the best parameterization to use at each time. This study presents a methodology to progressively refine the design space, which combines parametric effectiveness with a differential evolutionary algorithm in order to create an optimal parameterization. In this manuscript, we show that by doing the parameterization at the CAD level, we can impose higher level constraints on the shape, such as the axial chord length, the trailing edge radius and G2 geometric continuity between the suction side and pressure side at the leading edge. Additionally, the adjoint sensitivities are filtered out and only smooth shapes are produced during the optimization process. The use of algorithmic differentiation for the CAD kernel and grid generator allows computing the grid sensitivities to machine accuracy and avoid the limited arithmetic precision and the truncation error of finite differences. Then, the parametric effectiveness is computed to rate the ability of a set of CAD design parameters to produce the design shape change dictated by the adjoint sensitivities. During the optimization process, the design space is progressively enlarged using the knot insertion algorithm which allows introducing new control points whilst preserving the initial shape. The position of the inserted knots is generally assumed. However, this assumption can hinder the creation of better parameterizations that would allow producing more localized shape changes where the adjoint sensitivities dictate. To address this, we propose using a differential evolutionary algorithm to maximize the parametric effectiveness by optimizing the location of the inserted knots. This allows the optimizer to gradually explore larger design spaces and to use an optimal CAD-based parameterization during the course of the optimization. The method is tested on the LS89 turbine cascade and large aerodynamic improvements in the entropy generation are achieved whilst keeping the exit flow angle fixed. The trailing edge and axial chord length, which are kept fixed as manufacturing constraints. The optimization results show that the multilevel optimizations were more efficient than the single level optimization, even though they used the same number of design variables at the end of the multilevel optimizations. Furthermore, the multilevel optimization where the parameterization is created using the optimal knot positions results in a more efficient strategy to reach a better optimum than the multilevel optimization where the position of the knots is arbitrarily assumed.

Keywords: adjoint, CAD, knots, multilevel, optimization, parametric effectiveness

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Authors: N. Manjunatha, K. T. Rangswamy, N. Nagaraju, H. A. Prameela, P. Rudraswamy, M. Krishnareddy

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The identification of virus in cowpea especially potyviruses is confusing. Even though there are several studies on viruses causing diseases in cowpea, difficult to distinguish based on symptoms and serological detection. The differentiation of potyviruses considering as a constraint, the present study is initiated for molecular characterization, host plant resistance and epidemiology of the BCMV infecting cowpea. The etiological agent causing cowpea mosaic was identified as Bean Common Mosaic Virus (BCMV) on the basis of RT-PCR and electron microscopy. An approximately 750bp PCR product corresponding to coat protein (CP) region of the virus and the presence of long flexuous filamentous particles measuring about 952 nm in size typical to genus potyvirus were observed under electron microscope. The characterized virus isolate genome had 10054 nucleotides, excluding the 3’ terminal poly (A) tail. Comparison of polyprotein of the virus with other potyviruses showed similar genome organization with 9 cleavage sites resulted in 10 functional proteins. The pairwise sequence comparison of individual genes, P1 showed most divergent, but CP gene was less divergent at nucleotide and amino acid level. A phylogenetic tree constructed based on multiple sequence alignments of the polyprotein nucleotide and amino acid sequences of cowpea BCMV and potyviruses showed virus is closely related to BCMV-HB. Whereas, Soybean variant of china (KJ807806) and NL1 isolate (AY112735) showed 93.8 % (5’UTR) and 94.9 % (3’UTR) homology respectively with other BCMV isolates. This virus transmitted to different leguminous plant species and produced systemic symptoms under greenhouse conditions. Out of 100 cowpea genotypes screened, three genotypes viz., IC 8966, V 5 and IC 202806 showed immune reaction in both field and greenhouse conditions. Single marker analysis (SMA) was revealed out of 4 SSR markers linked to BCMV resistance, M135 marker explains 28.2 % of phenotypic variation (R2) and Polymorphic information content (PIC) value of these markers was ranged from 0.23 to 0.37. The correlation and regression analysis showed rainfall, and minimum temperature had significant negative impact and strong relationship with aphid population, whereas weak correlation was observed with disease incidence. Path coefficient analysis revealed most of the weather parameters exerted their indirect contributions to the aphid population and disease incidence except minimum temperature. This study helps to identify specific gaps in knowledge for researchers who may wish to further analyse the science behind complex interactions between vector-virus and host in relation to the environment. The resistant genotypes identified are could be effectively used in resistance breeding programme.

Keywords: cowpea, epidemiology, genotypes, virus

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Authors: Batul Kagalwala

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The nervous system is made up of complex delicate structures such as the spinal cord, peripheral nerves and the brain. These are prone to various types of injury ranging from neurodegenerative diseases to trauma leading to diseases like Parkinson's, Alzheimer's, multiple sclerosis, amyotrophic lateral sclerosis (ALS), multiple system atrophy etc. Unfortunately, because of the complicated structure of nervous system, spontaneous regeneration, repair and healing is seldom seen due to which brain damage, peripheral nerve damage and paralysis from spinal cord injury are often permanent and incapacitating. Hence, innovative and standardized approach is required for advance treatment of neurological injury. Nigella sativa (N. sativa), an annual flowering plant native to regions of southern Europe and Asia; has been suggested to have neuroprotective and anti-seizures properties. Neuroregeneration is found to occur in damaged cells when treated using extract of N. sativa. Due to its proven health benefits, lots of experiments are being conducted to extract all the benefits from the plant. The flowers are delicate and are usually pale blue and white in color with small black seeds. These seeds are the source of active components such as 30–40% fixed oils, 0.5–1.5% essential oils, pharmacologically active components containing thymoquinone (TQ), ditimoquinone (DTQ) and nigellin. In traditional medicine, this herb was identified to have healing properties and was extensively used Middle East and Far East for treating diseases such as head ache, back pain, asthma, infections, dysentery, hypertension, obesity and gastrointestinal problems. Literature studies have confirmed the extract of N. sativa seeds and TQ have inhibitory effects on inducible nitric oxide synthase and production of nitric oxide as well as anti-inflammatory and anticancer activities. Experimental investigation will be conducted to understand which ingredient of N. sativa causes neuroregeneration and roots to its healing property. An aqueous/ alcoholic extract of N. sativa will be made. Seed oil is also found to have used by researchers to prepare such extracts. For the alcoholic extracts, the seeds need to be powdered and soaked in alcohol for a period of time and the alcohol must be evaporated using rotary evaporator. For aqueous extracts, the powder must be dissolved in distilled water to obtain a pure extract. The mobile phase will be the extract while the suitable stationary phase (substance that is a good adsorbent e.g. silica gels, alumina, cellulose etc.) will be selected. Different ingredients of N. sativa will be separated using High Performance Liquid Chromatography (HPLC) for treating damaged cells. Damaged brain cells will be treated individually and in different combinations of 2 or 3 compounds for different intervals of time. The most suitable compound or a combination of compounds for the regeneration of cells will be determined using DOE methodology. Later the gene will also be determined and using Polymerase Chain Reaction (PCR) it will be replicated in a plasmid vector. This plasmid vector shall be inserted in the brain of the organism used and replicated within. The gene insertion can also be done by the gene gun method. The gene in question can be coated on a micro bullet of tungsten and bombarded in the area of interest and gene replication and coding shall be studied. Investigation on whether the gene replicates in the organism or not will be examined.

Keywords: black cumin, brain cells, damage, extract, neuroregeneration, PCR, plasmids, vectors

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Authors: Nadezhda F. Mikahailova, Margarita E. Fattakhova, Mirgarita A. Mironova, Ekaterina V. Vyacheslavova, Vladimir A. Mikahailov

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Stigma is a significant obstacle to the successful adaptation of deaf students to the conditions of an educational institution, especially for those who study in inclusion. The aim of the study was to identify the spheres of life which are the most significant for developing of the stigma of deaf students; to assess the influence of factors associated with deafness on the degree of their self-stigmatization (time and degree of hearing loss, type of education - inclusion / differentiation) and to find out who is more prone to stigma - which characteristics of personality, identity, mental health and coping are specific for those deaf who demonstrates stigmatizing attitudes. The study involved 154 deaf and hard-of-hearing students (85 male and 69 female) aged from 18 to 45 years - 28 students of the Herzen State Pedagogical University (St. Petersburg), who study in inclusion, 108 students of the National Research Technological University and 18 students of the Aviation Technical College (Kazan) - students in groups with a sign language interpreter. We used the following methods: modified questionnaire 'Self-assessment and coping strategies' (Jambor & Elliot, 2005), Scale of self-esteem (Rosenberg et al, 1995), 'Big-Five' (Costa&McCrae, 1997), TRF (Becker, 1989), WCQ (Lazarus & Folkman, 1988), self-stigma scale (Mikhailov, 2008). The severity of self-stigmatization of deaf and hard of hearing students was determined by the degree of deafness and the time they live with hearing loss, learning conditions, the type of self-identification (acculturation), personality traits, and the specifics of coping behavior. Persons with congenital hearing loss more often noted a benevolent and sympathetic attitude towards them on the part of the hearers and less often, due to deafness, limited themselves to visiting public places than late deaf people, which indicates 'get rid of' the experience of their defect and normalization of the state. Students studying in conditions of inclusion more often noted the dismissive attitude of society towards deaf people. Individuals with mild to moderate hearing loss were more likely to fear marriage and childbearing because of their deafness than students with profound hearing loss. Those who considered themselves disabled (49% of all respondents) were more inclined to cope with seeking social support and less used 'distancing' coping. Those who believed that their quality of life and social opportunities were most influenced by the attitude of society towards the deaf (39%) were distinguished by a less pronounced sense of self-worth, a desire for autonomy, and frequent usage of 'avoidance' coping strategies. 36.4% of the respondents noted that there have been situations in their lives when people learned that they are deaf, began to treat them worse. These respondents had predominantly deaf acculturation, but more often, they used 'bicultural skills,' specific coping for the deaf, and had a lower level of extraversion and emotional stability. 31.2% of the respondents tried to hide from others that they have hearing problems. They considered themselves to be in a culture of hearing, used coping strategies 'bicultural skills,' and had lower levels of extraversion, cooperation, and emotional stability. Acknowledgment: Supported by the RFBR № 19-013-0040

Keywords: acculturation, coping, deafness, stigmatization

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Authors: Helen Bartsch, Markus Feldmann

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The dimensioning of steel structures subject to fatigue loads, such as wind turbines, bridges, masts and towers, crane runways and weirs or components in crane construction, is often dominated by fatigue verification. The fatigue details defined by the welded connections, such as butt or cruciform joints, longitudinal welds, welded-on or welded-in stiffeners, etc., are decisive. In Europe, the verification is usually carried out according to EN 1993-1-9 on a nominal stress basis. The basis is the detailed catalog, which specifies the fatigue strength of the various weld and construction details according to fatigue classes. Until now, a relation between fatigue classes and weld imperfection sizes is not included. Quality levels for imperfections in fusion-welded joints in steel, nickel, titanium and their alloys are regulated in EN ISO 5817, which, however, doesn’t contain direct correlations to fatigue resistances. The question arises whether some imperfections might be tolerable to a certain extent since they may be present in the test data used for detail classifications dating back decades ago. Although current standardization requires proof of satisfying limits of imperfection sizes, it would also be possible to tolerate welds with certain irregularities if these can be reliably quantified by non-destructive testing. Fabricators would be prepared to undertake carefully and sustained weld inspection in view of the significant economic consequences of such unfavorable fatigue classes. This paper presents investigations on the fatigue behavior of common welded details containing imperfections. In contrast to the common nominal stress concept, local fatigue concepts were used to consider the true stress increase, i.e., local stresses at the weld toe and root. The actual shape of a weld comprising imperfections, e.g., gaps or undercuts, can be incorporated into the fatigue evaluation, usually on a numerical basis. With the help of the effective notch stress concept, the fatigue resistance of detailed local weld shapes is assessed. Validated numerical models serve to investigate notch factors of fatigue details with different geometries. By utilizing parametrized ABAQUS routines, detailed numerical studies have been performed. Depending on the shape and size of different weld irregularities, fatigue classes can be defined. As well load-carrying welded details, such as the cruciform joint, as non-load carrying welded details, e.g., welded-on or welded-in stiffeners, are regarded. The investigated imperfections include, among others, undercuts, excessive convexity, incorrect weld toe, excessive asymmetry and insufficient or excessive throat thickness. Comparisons of the impact of different imperfections on the different types of fatigue details are made. Moreover, the influence of a combination of crucial weld imperfections on the fatigue resistance is analyzed. With regard to the trend of increasing efficiency in steel construction, the overall aim of the investigations is to include a more economical differentiation of fatigue details with regard to tolerance sizes. In the long term, the harmonization of design standards, execution standards and regulations of weld imperfections is intended.

Keywords: effective notch stress, fatigue, fatigue design, weld imperfections

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Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

Procedia PDF Downloads 192

Authors: Md. M. Hossain, Regan Roat, Jenica Christopherson, Colette Free, Zhiguang Guo

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Long noncoding RNA (lncRNA) mediated post-transcriptional gene regulation, and their epigenetic landscapes have been shown to be involved in many human diseases. However, their regulation in diabetes through governing islet’s β-cell function and survival needs to be elucidated. Due to the technical and ethical constraints, it is difficult to study their role in β-cell function and survival in human under in vivo condition. In this study, humanized mice have been developed through transplanting human pancreatic islet under the kidney capsule of NOD.SCID mice and induced β-cell death leading to diabetes condition to study lncRNA mediated regulation. For this, human islets from 3 donors (3000 IEQ, purity > 80%) were transplanted under the kidney capsule of STZ induced diabetic NOD.scid mice. After at least 2 weeks of normoglycecemia, lymphocytes from diabetic NOD mice were adoptively transferred and islet grafts were collected once blood glucose reached > 200 mg/dl. RNA from human donor islets, islet grafts from humanized mice with either adoptive lymphocyte transfer (ALT) or PBS control (CTL) were ribodepleted; barcoded fragment libraries were constructed and sequenced on the Ion Proton sequencer. lncRNA expression in isolated human islets, islet grafts from humanized mice with and without induced β-cell death and their regulation in human islets function in vitro under glucose challenge, cytokine mediated inflammation and induced apoptotic condition were investigated. Out of 3155 detected lncRNAs, 299 that highly expressed in islets were found to be significantly downregulated and 224 upregulated in ALT compared to CTL. Most of these are found to be collocated within 5 kb upstream and 1 kb downstream of 788 up- and 624 down-regulated mRNAs. Genomic Regions Enrichment of Annotations Analysis revealed deregulated and collocated genes are related to pancreas endocrine development; insulin synthesis, processing, and secretion; pancreatitis and diabetes. Many of them, that found to be located within enhancer domains for islet specific gene activity, are associated to the deregulation of known islet/βcell specific transcription factors and genes that are important for β-cell differentiation, identity, and function. RNA sequencing analysis revealed aberrant lncRNA expression which is associated to the deregulated mRNAs in β-cell function as well as in molecular pathways related to diabetes. A distinct set of candidate lncRNA isoforms were identified as highly enriched and specific to human islets, which are deregulated in human islets from donors with different BMIs and with type 2 diabetes. These RNAs show an interesting regulation in cultured human islets under glucose stimulation and with induced β-cell death by cytokines. Aberrant expression of these lncRNAs was detected in the exosomes from the media of islets cultured with cytokines. Results of this study suggest that the islet specific lncRNAs are deregulated in human islet with β-cell death, hence important in diabetes. These lncRNAs might be important for human β-cell function and survival thus could be used as biomarkers and novel therapeutic targets for diabetes.

Keywords: β-cell, humanized mouse, pancreatic islet, LncRNAs

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Authors: Jan F. Talts, Amit Saxena, Kåre Engkilde

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Chemotherapy-induced peripheral neuropathy (CIPN) is one of the most frequent side effects caused by anti-neoplastic agents, with a prevalence from 19 % to 85 %. Clinically, CIPN is a mostly sensory neuropathy leading to pain and to motor and autonomic changes. Due to its high prevalence among cancer patients, CIPN constitutes a major problem for both cancer patients and survivors, especially because currently, there is no single effective method of preventing CIPN. Hearing loss is the most common form of sensory impairment in humans and can be caused by ototoxic chemical compounds such as chemotherapy (platinum-based antineoplastic agents).In rodents, single or repeated cisplatin injections induce peripheral neuropathy and hearing impairment mimicking human disorder, allowing studying the efficacy of new pharmacological candidates in chemotherapy-induced hearing loss and peripheral neuropathy. RNA sequencing data from full term amniotic fluid (TAF) mesenchymal stemcell (MSC) clones was used to identify neural-specific markers present on TAF-MSC. Several prospective neural markers were tested by flow cytometry on cultured TAF-MSC. One of these markers was used for cell-sorting using Tyto MACSQuant cell sorter, and the neural marker positive cell population was expanded for several passages to the final therapeutic product stage. Peripheral neuropathy and hearing loss was induced in mice by administration of cisplatin in three week-long cycles. The efficacy of neural-specific TAF-MSC in treating hearing loss and pain perception was evaluated by administration of three injections of 3 million cells/kg by intravenous route or three injections of 3 million cells/kg by intra-arterial route after each cisplatin cycle treatment. Auditory brainstem responses (ABR) are electric potentials recorded from scalp electrodes, and the first ABR wave represents the summed activity of the auditory nerve fibers contacting the inner hair cells. For ABR studies, mice were anesthetized, then earphones were placed in the left ear of each mouse, an active electrode was placed in the vertex of the skull, a reference electrode under the skin of the mastoid bone, and a ground electrode in the neck skin. The stimuli consisted of tone pips of five frequencies (2, 4, 6, 12, 16, and 24 kHz) at various sound levels (from 0 to 90 dB) ranging to cover the mouse auditory frequency range. The von Frey test was used to assess the onset and maintenance of mechanical allodynia over time. Mice were placed in clear plexiglass cages on an elevated mesh floor and tested after 30 min of habituation. Mechanical paw withdrawal threshold was examined using an electronic von Frey anesthesiometer. Cisplatin groups treated with three injections of 3 million cells/kg by intravenous route and three injections of 3 million cells/kg by intra-arterial route after each cisplatin cycle treatment presented, a significant increase of hearing acuity characterized by a decrease of ABR threshold and a decrease of neuropathic pain characterized by an increase of von Frey paw withdrawal threshold compared to controls only receiving cisplatin. This study shows that treatment with MSCselected for neural specificity presents significant positive efficacy on the chemotherapy-induced neuropathic pain and the chemotherapy-induced hearing loss.

Keywords: mesenchymal stem cell, peripheral neuropathy, amniotic fluid, regenerative medicine

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Authors: Ana Paula S. Horodenski, Priscila Pelegrini, Salli Baggenstoss

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The brewing industry with artisanal concepts seeks to serve a specific market, with diversified production that has been gaining ground in the national environment, also in the Amazon region. This growth is due to the more demanding consumer, with a diversified taste that wants to try new types of beer, enjoying products with new aromas, flavors, as a differential of what is so widely spread through the big industrial brands. Thus, through qualitative research methods, the study aimed to investigate how is the process of managing the production of a craft brewery in a city in the northern State of Mato Grosso (BRAZIL), providing knowledge of production processes and strategies in the industry. With the efficient use of resources, it is possible to obtain the necessary quality and provide better performance and differentiation of the company, besides analyzing the best management model. The research is descriptive with a qualitative approach through a case study. For the data collection, a semi-structured interview was elaborated, composed of the areas: microbrewery characterization, artisan beer production process, and the company supply chain management. Also, production processes were observed during technical visits. With the study, it was verified that the artisan brewery researched develops preventive maintenance strategies with the inputs, machines, and equipment, so that the quality of the product and the production process are achieved. It was observed that the distance from the supplying centers makes the management of processes and the supply chain be carried out with a longer planning time so that the delivery of the final product is satisfactory. The production process of the brewery is composed of machines and equipment that allows the control and quality of the product, which the manager states that for the productive capacity of the industry and its consumer market, the available equipment meets the demand. This study also contributes to highlight one of the challenges for the development of small breweries in front of the market giants, that is, the legislation, which fits the microbreweries as producers of alcoholic beverages. This makes the micro and small business segment to be taxed as a major, who has advantages in purchasing large batches of raw materials and tax incentives because they are large employers and tax pickers. It was possible to observe that the supply chain management system relies on spreadsheets and notes that are done manually, which could be simplified with a computer program to streamline procedures and reduce risks and failures of the manual process. In relation to the control of waste and effluents affected by the industry is outsourced and meets the needs. Finally, the results showed that the industry uses preventive maintenance as a productive strategy, which allows better conditions for the production and quality of artisanal beer. The quality is directly related to the satisfaction of the final consumer, being prized and performed throughout the production process, with the selection of better inputs, the effectiveness of the production processes and the relationship with the commercial partners.

Keywords: artisanal brewery, production management, production processes, supply chain

Procedia PDF Downloads 94

Authors: A. V. Harutyunyan

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The investigation results of rocks at high pressures and temperatures have revealed the intervals of changes of seismic waves and density, as well as some processes taking place in rocks. In the serpentinized rocks, as a consequence of dehydration, abrupt changes in seismic waves and density have been recorded. Hydrogen-bearing components are released which combine with carbon-bearing components. As a result, hydrocarbons formed. The investigated samples are smelted. Then, geofluids and hydrocarbons migrate into the upper horizons of the Earth crust by the deep faults. Then their differentiation and accumulation in the jointed rocks of the faults and in the layers with collecting properties takes place. Under the majority of the hydrocarbon deposits, at a certain depth, magmatic centers and deep faults are recorded. The investigation results of the serpentinized rocks with numerous geological-geophysical factual data allow understanding that hydrocarbons are mainly formed in both the offshore part of the ocean and at different depths of the continental crust. Experiments have also shown that the dehydration of the serpentinized rocks is accompanied by an explosion with the instantaneous increase in pressure and temperature and smelting the studied rocks. According to numerous publications, hydrocarbons and diamonds are formed in the upper part of the mantle, at the depths of 200-400km, and as a consequence of geodynamic processes, they rise to the upper horizons of the Earth crust through narrow channels. However, the genesis of metamorphogenic diamonds and the diamonds found in the lava streams formed within the Earth crust, remains unclear. As at dehydration, super high pressures and temperatures arise. It is assumed that diamond crystals are formed from carbon containing components present in the dehydration zone. It can be assumed that besides the explosion at dehydration, secondary explosions of the released hydrogen take place. The process is naturally accompanied by seismic phenomena, causing earthquakes of different magnitudes on the surface. As for the diamondiferous kimberlites, it is well-known that the majority of them are located within the ancient shield and platforms not obligatorily connected with the deep faults. The kimberlites are formed at the shallow location of dehydrated masses in the Earth crust. Kimberlites are younger in respect of containing ancient rocks containing serpentinized bazites and ultrbazites of relicts of the paleooceanic crust. Sometimes, diamonds containing water and hydrocarbons showing their simultaneous genesis are found. So, the geofluids, hydrocarbons and diamonds, according to the new concept put forward, are formed simultaneously from serpentinized rocks as a consequence of their dehydration at different depths of the Earth crust. Based on the concept proposed by us, we suggest discussing the following: -Genesis of gigantic hydrocarbon deposits located in the offshore area of oceans (North American, Mexican Gulf, Cuanza-Kamerunian, East Brazilian etc.) as well as in the continental parts of different mainlands (Kanadian-Arctic Caspian, East Siberian etc.) - Genesis of metamorphogenic diamonds and diamonds in the lava streams (Guinea-Liberian, Kokchetav, Kanadian, Kamchatka-Tolbachinian, etc.).

Keywords: dehydration, diamonds, hydrocarbons, serpentinites

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Authors: M. D. Nirmani, K. L. N. Perera, G. H. Galhena

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Dengue has become one of the most important arbo-viral disease in all tropical and subtropical regions of the world. Aedes aegypti, is the principal vector of the virus, vary in both epidemiological and behavioral characteristics, which could be finely measured through DNA sequence comparison at their population level. Such knowledge in the population differences can assist in implementation of effective vector control strategies allowing to make estimates of the gene flow and adaptive genomic changes, which are important predictors of the spread of Wolbachia infection or insecticide resistance. As such, this study was undertaken to investigate the phylogenetic relationships of Ae. aegypti from Galle and Colombo, Sri Lanka, based on the ribosomal protein region which spans between two exons, in order to understand the geographical distribution of genetically distinct mosquito clades and its impact on mosquito control measures. A 320bp DNA region spanning from 681-930 bp, corresponding to the ribosomal protein, was sequenced in 62 Ae. aegypti larvae collected from Galle (N=30) and Colombo (N=32), Sri Lanka. The sequences were aligned using ClustalW and the haplotypes were determined with DnaSP 5.10. Phylogenetic relationships among haplotypes were constructed using the maximum likelihood method under Tamura 3 parameter model in MEGA 7.0.14 including three previously reported sequences of Australian (N=2) and Brazilian (N=1) Ae. aegypti. The bootstrap support was calculated using 1000 replicates and the tree was rooted using Aedes notoscriptus (GenBank accession No. KJ194101). Among all sequences, nineteen different haplotypes were found among which five haplotypes were shared between 80% of mosquitoes in the two populations. Seven haplotypes were unique to each of the population. Phylogenetic tree revealed two basal clades and a single derived clade. All observed haplotypes of the two Ae. aegypti populations were distributed in all the three clades, indicating a lack of genetic differentiation between populations. The Brazilian Ae. aegypti haplotype and one of the Australian haplotypes were grouped together with the Sri Lankan basal haplotype in the same basal clade, whereas the other Australian haplotype was found in the derived clade. Phylogram showed that Galle and Colombo Ae. aegypti populations are highly related to each other despite the large geographic distance (129 Km) indicating a substantial genetic similarity between them. This may have probably arisen from passive migration assisted by human travelling and trade through both land and water as the two areas are bordered by the sea. In addition, studied Sri Lankan mosquito populations were closely related to Australian and Brazilian samples. Probably this might have caused by shipping industry between the three countries as all of them are fully or partially enclosed by sea. For example, illegal fishing boats migrating to Australia by sea is perhaps a good mean of transportation of all life stages of mosquitoes from Sri Lanka. These findings indicate that extensive mosquito migrations occur between populations not only within the country, but also among other countries in the world which might be a main barrier to the successful vector control measures.

Keywords: Aedes aegypti, dengue control, extensive mosquito migration, haplotypes, phylogeny, ribosomal protein

Procedia PDF Downloads 153

Authors: Maryam Ziaee

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This research proposes a new approach and provides an opportunity for manufacturing companies to produce large amounts of products that meet their prospective customers’ tastes, needs, and expectations and simultaneously enable manufacturers to increase their profit. Mass customization is the production of products or services to meet each individual customer’s desires to the greatest possible extent in high quantities and at reasonable prices. This process takes place at different levels such as the customization of goods’ design, assembly, sale, and delivery status, and classifies in several categories. The main focus of this study is on one class of mass customization, called optional customization, in which companies try to provide their customers with as many options as possible to customize their products. These options could range from the design phase to the manufacturing phase, or even methods of delivery. Mass customization values customers’ tastes, but it is only one side of clients’ satisfaction; on the other side is companies’ fast responsiveness delivery. It brings the concept of agility, which is the ability of a company to respond rapidly to changes in volatile markets in terms of volume and variety. Indeed, mass customization is not effectively feasible without integrating the concept of agility. To gain the customers’ satisfaction, the companies need to be quick in responding to their customers’ demands, thus highlighting the significance of agility. This research offers a different method that successfully integrates mass customization and fast production in manufacturing industries. This research is built upon the hypothesis that the success key to being agile in mass customization is to forecast demand, cooperate with suppliers, and control inventory. Therefore, the significance of the supply chain (SC) is more pertinent when it comes to this stage. Since SC behavior is dynamic and its behavior changes constantly, companies have to apply one of the predicting techniques to identify the changes associated with SC behavior to be able to respond properly to any unwelcome events. System dynamics utilized in this research is a simulation approach to provide a mathematical model among different variables to understand, control, and forecast SC behavior. The final stage is delayed differentiation, the production strategy considered in this research. In this approach, the main platform of products is produced and stocked and when the company receives an order from a customer, a specific customized feature is assigned to this platform and the customized products will be created. The main research question is to what extent applying system dynamics for the prediction of SC behavior improves the agility of mass customization. This research is built upon a qualitative approach to bring about richer, deeper, and more revealing results. The data is collected through interviews and is analyzed through NVivo software. This proposed model offers numerous benefits such as reduction in the number of product inventories and their storage costs, improvement in the resilience of companies’ responses to their clients’ needs and tastes, the increase of profits, and the optimization of productivity with the minimum level of lost sales.

Keywords: agility, manufacturing, resilience, supply chain

Procedia PDF Downloads 66

Authors: A. Moghadasein, M. Ghasemi, S. Fazelifar

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Aim: Chemerin is a novel adipokine that play an important role in regulating lipid metabolism and abiogenesis. Chemerin is dependent on autocrine and paracrine signals for the differentiation and maturation of fat cells; it also regulates glucose uptake in fat cells and stimulates lipolysis. It has been reported that in adipocytes, chemerin enhances the insulin-stimulated glucose and causes the phosphorylation of tyrosine in Insulin receptor substrate. According to the studies, Chemerin may increase insulin sensitivity in adipose tissue and is largely associated with Body mass index, triglycerides, and blood pressure in those with normal glucose tolerance. There is limited information available regarding the effect of exercise training on serum chemerin concentrations. The purpose of this study was to investigate the effect of endurance training on serum chemerin levels and lipids of plasma in overweight women. Methodology: This study was a quasi-experimental research with a pre-post test design. After required examination and verification of high pressure by the physician, 22 obese subjects (age: 35.64±5.55 yr, weight: 75.62±9.30 kg, body mass index: 32.4±1.6 kg/m2) were randomly assigned to aerobic training (n= 12) and control (n= 12) groups. Participants completed a questionnaire indicating the lack of sports history during the past six months, the lack of anti-hypertension drugs use, hormone therapy, cardiovascular problems, and complete stoppage of menstrual cycle. Aerobic training was performed 3 times weekly for 8 weeks. Resting levels of chemerin plasma, metabolic parameters were measured prior to and after the intervention. The control group did not participate in any training program. In this study, ethical considerations included the complete description of the objectives to the study participants, ensuring the confidentiality of their information. Kolmogorov-Smirnov and Levin test were used for determining the normal distribution of data and homogeneity of variances, respectively. Analyze of variance with repeated measure were used to investigate the changes in the intra-group and the differences in inter-group of variables. Statistical operations were performed using SPSS 16 and the significance level of the tests was considered at P < 0.05. Results: After an 8 week aerobic training, levels of chemerin plasma were significantly decreased in aerobic trained group when compared with their control groups (p < 0.05).Concurrently, levels of HDL-c were significantly decreased (p < 0.05) whereas, levels of cholesterol, TG and LDL-c, showed no significant changes (p > 0.05). No significant correlations between chemerin levels and weight loss were observed in subjects with overweight women. Conclusion: The present study demonstrated, 8 weeks aerobic training, reduced serum chemerin concentrations in overweight women. Whereas, aerobic training exercise programmers affected the lipid profile response of obese subjects differently. However further research is warranted in order to unravel the molecular mechanism for the range of responses and the role of serum chemerin.

Keywords: chemerin, aerobic training, lipid profile, obese women

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Authors: Ashish Thakur

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This technical paper was written in view of focusing the biomaterials and its various applications in modern industries. Author tires to elaborate not only the medical, infect plenty of application in other industries. The scope of the research area covers the wide range of physical, biological and chemical sciences that underpin the design of biomaterials and the clinical disciplines in which they are used. A biomaterial is now defined as a substance that has been engineered to take a form which, alone or as part of a complex system, is used to direct, by control of interactions with components of living systems, the course of any therapeutic or diagnostic procedure. Biomaterials are invariably in contact with living tissues. Thus, interactions between the surface of a synthetic material and biological environment must be well understood. This paper reviews the benefits and challenges associated with surface modification of the metals in biomedical applications. The paper also elaborates how the surface characteristics of metallic biomaterials, such as surface chemistry, topography, surface charge, and wettability, influence the protein adsorption and subsequent cell behavior in terms of adhesion, proliferation, and differentiation at the biomaterial–tissue interface. The chapter also highlights various techniques required for surface modification and coating of metallic biomaterials, including physicochemical and biochemical surface treatments and calcium phosphate and oxide coatings. In this review, the attention is focused on the biomaterial-associated infections, from which the need for anti-infective biomaterials originates. Biomaterial-associated infections differ markedly for epidemiology, aetiology and severity, depending mainly on the anatomic site, on the time of biomaterial application, and on the depth of the tissues harbouring the prosthesis. Here, the diversity and complexity of the different scenarios where medical devices are currently utilised are explored, providing an overview of the emblematic applicative fields and of the requirements for anti-infective biomaterials. In addition to this, chapter introduces nanomedicine and the use of both natural and synthetic polymeric biomaterials, focuses on specific current polymeric nanomedicine applications and research, and concludes with the challenges of nanomedicine research. Infection is currently regarded as the most severe and devastating complication associated to the use of biomaterials. Osteoporosis is a worldwide disease with a very high prevalence in humans older than 50. The main clinical consequences are bone fractures, which often lead to patient disability or even death. A number of commercial biomaterials are currently used to treat osteoporotic bone fractures, but most of these have not been specifically designed for that purpose. Many drug- or cell-loaded biomaterials have been proposed in research laboratories, but very few have received approval for commercial use. Polymeric nanomaterial-based therapeutics plays a key role in the field of medicine in treatment areas such as drug delivery, tissue engineering, cancer, diabetes, and neurodegenerative diseases. Advantages in the use of polymers over other materials for nanomedicine include increased functionality, design flexibility, improved processability, and, in some cases, biocompatibility.

Keywords: nanomedicine, tissue, infections, biomaterials

Procedia PDF Downloads 235

Authors: K. Mohon Singh

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Cambro-Ordovician (512.5 ± 8.7 Ma) felsic magmatism in the Kyrdem region of Meghalaya plateau, herewith referred to as Kyrdem granitoids (KG), intrudes the low-grade Shillong Group of metasediments and Precambrian Basement Gneissic complex forming an oval-shaped plutonic body with longer axis almost trending N-S. Thermal aureole is poorly developed or covered under the alluvium. KG exhibit very coarse grained porphyritic texture with abundant K-feldspar megacrysts (up to 9cm long) and subordinate amount of amphibole, biotite, plagioclase, and quartz. The size of K-feldspar megacrysts increases from margin (Dwarksuid) to the interior (Kyrdem) of the KG pluton. Late felsic pulses as fine grained granite, leucocratic (aplite), and pegmatite veins intrude the KG at several places. Grey and pink varieties of KG can be recognized, but pink colour of KG is the result of post-magmatic fluids, which have not affected the magnetic properties of KG. Modal composition of KG corresponds to quartz monzonite, monzogranite, and granodiorite. KG has been geochemically characterized as metaluminous (I-type) to peraluminous (S-type) granitoids. The KG is characterized by development of variable attitude of primary foliations mostly marked along the margin of the pluton and is located at the proximity of Tyrsad-Barapani lineament. The KG contains country rock xenoliths (amphibolite, gneiss, schist, etc.) which are mostly confined to the margin of the pluton, and microgranular enclaves (ME) are hosted in the porphyritic variety of KG. Microgranular Enclaves (ME) in Kyrdem Granitoids are fine- to medium grained, mesocratic to melanocratic, phenocryst bearing or phenocryst-free, rounded to ellipsoidal showing typical magmatic textures. Mafic-felsic phenocrysts in ME are partially corroded and dissolved because of their involvement in magma-mixing event, and thus represent xenocrysts. Sharp to diffused contacts of ME with host Kyrdem Granitoids, fine grained nature and presence of acicular apatite in ME suggest comingling and undercooling of coeval, semi-solidified ME magma within partly crystalline felsic host magma. Geochemical features recognize the nature of ME (molar A/CNK=0.76-1.42) and KG (molar A/CNK =0.41-1.75) similar to hybrid-type formed by mixing of mantle-derived mafic and crustal-derived felsic magmas. Major and trace including rare earth elements variations of ME suggest the involvement of combined processes such as magma mixing, mingling and crystallization differentiation in the evolution of ME but KG variations appear primarily controlled by fractionation of plagioclase, hornblende biotite, and accessory phases. Most ME are partially to nearly re-equilibrate chemically with felsic host KG during magma mixing and mingling processes.

Keywords: geochemistry, Kyrdem Granitoids, microgranular enclaves, Northeast India

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Authors: Jessie Rapoza, Petr Moroshkin, Jimmy Xu

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Chirality is omnipresent, in nature, in life, and in the field of physics. One intriguing example is the homochirality that has remained a great secret of life. Another is the pairs of mirror-image molecules – enantiomers. They are identical in atomic composition and therefore indistinguishable in the scalar physical properties. Yet, they can be either therapeutic or toxic, depending on their chirality. Recent studies suggest a potential link between abnormal levels of certain D-amino acids and some serious health impairments, including schizophrenia, amyotrophic lateral sclerosis, and potentially cancer. Although indistinguishable in their scalar properties, the chirality of a molecule reveals itself in interaction with the surrounding of a certain chirality, or more generally, a broken mirror-symmetry. In this work, we report on a system for chiral molecule detection, in which the mirror-symmetry is doubly broken, first by asymmetric structuring a nanopatterned plasmonic surface than by the incidence of circularly polarized light (CPL). In this system, the incident circularly-polarized light induces a surface plasmon polariton (SPP) wave, propagating along the asymmetric plasmonic surface. This SPP field itself is chiral, evanescently bound to a near-field zone on the surface (~10nm thick), but with an amplitude greatly intensified (by up to 104) over that of the incident light. It hence probes just the molecules on the surface instead of those in the volume. In coupling to molecules along its path on the surface, the chiral SPP wave favors one chirality over the other, allowing for chirality detection via the change in an optical rectification current measured at the edges of the sample. The asymmetrically structured surface converts the high-frequency electron plasmonic-oscillations in the SPP wave into a net DC drift current that can be measured at the edge of the sample via the mechanism of optical rectification. The measured results validate these design concepts and principles. The observed optical rectification current exhibits a clear differentiation between a pair of enantiomers. Experiments were performed by focusing a 1064nm CW laser light at the sample - a gold grating microchip submerged in an approximately 1.82M solution of either L-arabinose or D-arabinose and water. A measurement of the current output was then recorded under both rights and left circularly polarized lights. Measurements were recorded at various angles of incidence to optimize the coupling between the spin-momentums of the incident light and that of the SPP, that is, spin-momentum locking. In order to suppress the background, the values of the photocurrent for the right CPL are subtracted from those for the left CPL. Comparison between the two arabinose enantiomers reveals a preferential signal response of one enantiomer to left CPL and the other enantiomer to right CPL. In sum, this work reports on the first experimental evidence of the feasibility of chiral molecule detection via optical rectification in a metal meta-grating. This nanoscale interfaced electrical detection technology is advantageous over other detection methods due to its size, cost, ease of use, and integration ability with read-out electronic circuits for data processing and interpretation.

Keywords: Chirality, detection, molecule, spin

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Authors: Leo Nnamdi Ozurumba-Dwight

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Messenger ribooxynucleic acid (mRNA) molecules are compositional, protein-based. These proteins, encoding mRNA molecules (which collectively connote the transcriptome), when analyzed by RNA sequencing (RNAseq), unveils the nature of gene expression in the RNA. The obtained gene expression provides clues of cellular traits and their dynamics in presentations. These can be studied in relation to function and responses. RNAseq is a practical concept in Genomics as it enables detection and quantitative analysis of mRNA molecules. Single cell and spatial transcriptomics both present varying avenues for expositions in genomic characteristics of single cells and pooled cells in disease conditions such as cancer, auto-immune diseases, hematopoietic based diseases, among others, from investigated biological tissue samples. Single cell transcriptomics helps conduct a direct assessment of each building unit of tissues (the cell) during diagnosis and molecular gene expressional studies. A typical technique to achieve this is through the use of a single-cell RNA sequencer (scRNAseq), which helps in conducting high throughput genomic expressional studies. However, this technique generates expressional gene data for several cells which lack presentations on the cells’ positional coordinates within the tissue. As science is developmental, the use of complimentary pre-established tissue reference maps using molecular and bioinformatics techniques has innovatively sprung-forth and is now used to resolve this set back to produce both levels of data in one shot of scRNAseq analysis. This is an emerging conceptual approach in methodology for integrative and progressively dependable transcriptomics analysis. This can support in-situ fashioned analysis for better understanding of tissue functional organization, unveil new biomarkers for early-stage detection of diseases, biomarkers for therapeutic targets in drug development, and exposit nature of cell-to-cell interactions. Also, these are vital genomic signatures and characterizations of clinical applications. Over the past decades, RNAseq has generated a wide array of information that is igniting bespoke breakthroughs and innovations in Biomedicine. On the other side, spatial transcriptomics is tissue level based and utilized to study biological specimens having heterogeneous features. It exposits the gross identity of investigated mammalian tissues, which can then be used to study cell differentiation, track cell line trajectory patterns and behavior, and regulatory homeostasis in disease states. Also, it requires referenced positional analysis to make up of genomic signatures that will be sassed from the single cells in the tissue sample. Given these two presented approaches to RNA transcriptomics study in varying quantities of cell lines, with avenues for appropriate resolutions, both approaches have made the study of gene expression from mRNA molecules interesting, progressive, developmental, and helping to tackle health challenges head-on.

Keywords: transcriptomics, RNA sequencing, single cell, spatial, gene expression.

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Authors: Yael Livneh, Shay Tzafrir, Ilan Meshoulam

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Technological startup companies have been recognized as bearing tremendous potential for business and economic success. However, many entrepreneurs who produce promising innovative ideas fail to implement them as successful businesses. A key argument for such failure is the entrepreneurs' lack of competence in adaptation of the relevant level of formality of human resource management (HRM). The purpose of the present research was to examine multiple antecedents and consequences of HRM formality in growing startup companies. A review of the research literature identified two central components of HRM formality: HR control and professionalism. The effect of three contextual predictors was examined. The first was an intra-organizational factor: the development level of the organization. We based on a differentiation between knowledge exploration and knowledge exploitation. At a given time, the organization chooses to focus on a specific mix of these orientations, a choice which requires an appropriate level of HRM formality, in order to efficiently overcome the challenges. It was hypothesized that the mix of orientations of knowledge exploration and knowledge exploitation would predict HRM formality. The second predictor was the personal characteristics the organization's leader. According the idea of blueprint effect of CEO's on HRM, it was hypothesized that the CEO's cognitive style would predict HRM formality. The third contextual predictor was an external organizational factor: the level of investor involvement. By using the agency theory, and based on Transaction Cost Economy, it was hypothesized that the level of investor involvement in general management and HRM would be positively related to the HRM formality. The effect of formality on trust was examined directly and indirectly by the mediation role of procedural justice. The research method included a time-lagged field study. In the first study, data was obtained using three questionnaires, each directed to a different source: CEO, HR position-holder and employees. 43 companies participated in this study. The second study was conducted approximately a year later. Data was recollected using three questionnaires by reapplying the same sample. 41 companies participated in the second study. The organizations samples included technological startup companies. Both studies included 884 respondents. The results indicated consistency between the two studies. HRM formality was predicted by the intra-organizational factor as well as the personal characteristics of the CEO, but not at all by the external organizational context. Specifically, the organizational orientations was the greatest contributor to both components of HRM formality. The cognitive style predicted formality to a lesser extent. The investor's involvement was found not to have any predictive effect on the HRM formality. The results indicated a positive contribution to trust in HRM, mainly via the mediation of procedural justice. This study contributed a new concept for technological startup company development by a mixture of organizational orientation. Practical implications indicated that the level of HRM formality should be matched to that of the company's development. This match should be challenged and adjusted periodically by referring to the organization orientation, relevant HR practices, and HR function characteristics. A relevant matching could enhance further trust and business success.

Keywords: control, formality, human resource management, organizational development, professionalism, technological startup company

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Authors: Elena I. Oprita, Oana Craciunescu, Rodica Tatia, Teodora Ciucan, Reka Barabas, Orsolya Raduly, Anca Oancea

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Healing of osteochondral defects requires repair of the damaged articular cartilage, the underlying subchondral bone and the interface between these tissues (the functional calcified layer). For this purpose, developing a single monophasic scaffold that can regenerate two specific lineages (cartilage and bone) becomes a challenge. The aim of this work was to develop variants of biodegradable cross-linked composite hydrogel based on natural polypeptides (gelatin), polysaccharides components (chondroitin-4-sulphate and hyaluronic acid), in a ratio of 2:0.08:0.02 (w/w/w) and mixed with Si-hydroxyapatite (Si-Hap), in two ratios of 1:1 and 2:1 (w/w). Si-Hap was synthesized and characterized as a better alternative to conventional Hap. Subsequently, both composite hydrogel variants were cross-linked with (N, N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with a small bioactive molecule (icariin). The small molecule icariin (Ica) (C33H40O15) is the main active constituent (flavonoid) of Herba epimedium used in traditional Chinese medicine to cure bone- and cartilage-related disorders. Ica enhances osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), facilitates matrix calcification and increases the specific extracellular matrix (ECM) components synthesis by chondrocytes. Afterward, the composite hydrogels were characterized for their physicochemical properties in terms of the enzymatic biodegradation in the presence of type I collagenase and trypsin, the swelling capacity and the degree of crosslinking (TNBS assay). The cumulative release of Ica and real-time concentration were quantified at predetermined periods of time, according to the standard curve of standard Ica, after hydrogels incubation in saline buffer at physiological parameters. The obtained cross-linked composite hydrogels enriched with small-molecule Ica were also characterized for morphology by scanning electron microscopy (SEM). Their cytocompatibility was evaluated according to EN ISO 10993-5:2009 standard for medical device testing. Thus, analyses regarding cell viability (Live/Dead assay), cell proliferation (Neutral Red assay) and cell adhesion to composite hydrogels (SEM) were performed using NCTC clone L929 cell line. The final results showed that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical, structural and biological properties to be used as a natural scaffold able to repair osteochondral defects. The data did not reveal any toxicity of composite hydrogels in NCTC stabilized cell lines within the tested range of concentrations. Moreover, cells were capable of spreading and proliferating on both composite hydrogel surfaces. In conclusion, the designed biodegradable cross-linked composites enriched with Si and Ica are recommended for further testing as natural temporary scaffolds, which can allow cell migration and synthesis of new extracellular matrix within osteochondral defects.

Keywords: composites, gelatin, osteochondral defect, small molecule

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Authors: Colin S. Chen, Chien-Jung Tien, Hsin-Jan Huang

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For weathering studies, the change of chemical constituents by biodegradation effect in diesel-contaminated soils are important factors to be considered, especially when there is a prolonged period of weathering processes. The objective was to evaluate biodegradation effects onto hydrocarbon fingerprinting and distribution patterns of diesel fuels, fuel source screening and differentiation, source-specific marker compounds, and diagnostic ratios of diesel fuel constituents by laboratory and field studies. Biodegradation processes of diesel contaminated soils were evaluated by experiments lasting for 15 and 12 months, respectively. The degradation of diesel fuel in top soils was affected by organic carbon content and biomass of microorganisms in soils. Higher depletion of total petroleum hydrocarbon (TPH), n-alkanes, and polynuclear aromatic hydrocarbons (PAHs) and their alkyl homologues was observed in soils containing higher organic carbon content and biomass. Decreased ratio of selected isoprenoids (i.e., pristane (Pr) and phytane (Ph)) including n-C17/pristane and n-C18/phytane was observed. The ratio of pristane/phytane was remained consistent for a longer period of time. At the end of the experimental period, a decrease of pristane/phytane was observed. Biomarker compounds of bicyclic sesquiterpanes (BS) were less susceptible to the effects of biodegradation. The ratios of characteristic factors such as C15 sesquiterpane/ 8β(H)-drimane (BS3/BS5), C15 sesquiterpane/ 8β(H)-drimane (BS4/BS5), 8β(H)-drimane/8β(H)-homodrimane (BS5/BS10), and C15 sesquiterpane/8β(H)-homodrimane (BS3/BS10) could be adopted for source identification of diesel fuels in top soil. However, for biodegradation processes lasted for six months but shorter than nine months, only BS3/BS5 and BS3/BS10 could be distinguished in two diesel fuels. In subsoil experiments (contaminated soil located 50 cm below), the ratios of characteristic factors including BS3/BS5, BS4/BS5, and BS5/BS10 were valid for source identification of two diesel fuels for nine month biodegradation. At the early stage of contamination, biomass of soil decreased significantly. However, 6 and 7 dominant species were found in soils in top soil experiments, respectively. With less oxygen and nutrients in subsoil, less biomass of microorganisms was observed in subsoils. Only 2 and 4 diesel-degrading species of microorganisms were identified in two soils, respectively. Parameters of double ratio such as fluorene/C1-fluorene: C2-phenanthrene/C3-phenanthrene (C0F/C1F:C2P/C3P) in both top and subsoil, C2-naphthalene/C2-phenanthrene: C1-phenanthrene/C3-phenanthrene (C2N/C2P:C1P/C3P), and C1-phenanthrene/C1-fluorene: C3-naphthalene/C3-phenanthrene (C1P/C1F:C3N/C3P) in subsoil could serve as forensic indicators in diesel contaminated sites. BS3/BS10:BS4/BS5 could be used in 6 to 9 months of biodegradation processes. Results of principal component analysis (PCA) indicated that source identification of diesel fuels in top soil could only be perofrmed for weathering process less than 6 months. For subsoil, identification can be conducted for weathering process less than 9 months. Ratio of isoprenoids (pristane and phytane) and PAHs might be affected by biodegradation in spilled sites. The ratios of bicyclic sesquiterpanes could serve as forensic indicators in diesel-contaminated soils. Finally, source identification was attemped for samples collected from different fuel contaminated sites by using the unique pattern of sesquiterpanes. It was anticipated that the information generated from this study would be adopted by decision makers to evaluate the liability of cleanup in diesel contaminated sites.

Keywords: biodegradation, diagnostic ratio, diesel fuel, environmental forensics

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Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

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Authors: Tsirekidze M., Aprasidze T.

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Introduction: The problem of dissocial behavior is at the heart of the social sciences and psychiatry; however, it should be noted that its psychiatric aspect is little studied, and some issues of the problem are still controversial. This is complicated by the diversity of terminological concepts in defining “dissocial behavior”, “behavioral disorder”, “abnormal behavior”, “deviant behavior”, “delinquent behavior”, etc. In literature, there is no comprehensive definition of the essence of dissociative behavior. Numerous attempts to systematize dissociative disorders should also be considered unsatisfactory, which is primarily related to the lack of solid criteria for defining this group of disorders. According to the clinical classification, dissocial behavior is divided into psychotic and non-psychotic forms. Such differentiation is conditional in nature since it is not always possible to draw precise, clear distinctions between these forms, and in addition, there is a transition of a behavior disorder or so-called intermediate forms. One group of authors distinguishes two main forms of deviant behavior in terms of both theoretical and practical significance - non-pathological and pathological. In recent years, especially, the non-pathological form of behavior disorder has become topical. It refers to a large group of forms of deviant behavior, the emergence of which is associated with psychologically full-fledged reactions of children and adolescents to stressful situations and extreme conditions. According to the authors, its concept is understandable-it is difficult to draw a line between psychologically understandable reactions and psychogenically induced reactive states. In addition, the concept of "normal" child and adolescent is, to some extent, a vague concept, as in medicine, any definition of the norm. From a practical (more precisely, pragmatic) point of view, the term "abnormal behavioral disorder" undoubtedly makes sense, especially for the purpose of forensic psychiatric examination. Non-pathological deviation mainly includes transient situational reactions, microsocial-pedagogical backwardness, and character accentuation.Deviant behavior was predominantly manifested in a non-pathological form, which, in our opinion, is due to the difficult socio-economic situation of the country, moral-ethical deprivation, and expressed frustration. By itself, society is an indicator of deviation. Add to this situation complicated factors such as micro-social-pedagogical leave, unfavorable family environment, and parenting defects. Consideration is also given to the connection of acceptable deviation with the personal structural features of the adolescent. Aim: The topic of our discussion is the dissocial behavior of the non-psychotic register. Methods: We surveyed 120 adolescents with deviant behaviors. 61% of them were diagnosed with various neuropsychiatric disorders. Results: Abnormal forms of deviant behavior were observed in 13%, and non-pathological forms in -69%. A combination of non-pathological and pathological forms was present in 10% of cases. In the case of non-pathological deviation, microsocial-pedagogical acceptance was revealed in 62%, character accentuation in 22%; during the pathological forms, pathological reactions were observed in 21%, and abnormal formation of the person -21%. Conclusion: It should be emphasized that in case of any of the above defects, if the so-called family psychosis, and medical and pedagogical habilitation measures for the adolescent, it is quite possible to prevent the abnormal development of the child's personality, correct his character, regulate behavior and develop positive labor-social relations.

Keywords: dissocial personality, deviant behavior, dissocial, delinquent behavior

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