Search results for: Tracey Bone

Authors: B. R. O. Omidiwura, A. F. Agboola, E. A. Iyayi

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Pure form of L-Dopa is used to enhance muscular development, fat breakdown and suppress Parkinson disease in humans. However, the L-Dopa in mucuna seed, when present with other antinutritional factors, causes nutritional disorders in monogastric animals. Information on the utilisation of pure L-Dopa in monogastric animals is scanty. Therefore, effect of L-Dopa on growth performance and carcass characteristics in broiler chickens was investigated. Two hundred and forty one-day-old chicks were allotted to six treatments, which consisted of a positive control (PC) with standard energy (3100Kcal/Kg) and negative control (NC) with high energy (3500Kcal/Kg). The rest 4 diets were NC+0.1, NC+0.2, NC+0.3 and NC+0.4% L-Dopa, respectively. All treatments had 4 replicates in a completely randomized design. Body weight gain, final weight, feed intake, dressed weight and carcass characteristics were determined. Body weight gain and final weight of birds fed PC were 1791.0 and 1830.0g, NC+0.1% L-Dopa were 1827.7 and 1866.7g and NC+0.2% L-Dopa were 1871.9 and 1910.9g respectively, and the feed intake of PC (3231.5g), were better than other treatments. The dressed weight at 1375.0g and 1357.1g of birds fed NC+0.1% and NC+0.2% L-Dopa, respectively, were similar but better than other treatments. Also, the thigh (202.5g and 194.9g) and the breast meat (413.8g and 410.8g) of birds fed NC+0.1% and NC+0.2% L-Dopa, respectively, were similar but better than birds fed other treatments. The drum stick of birds fed NC+0.1% L-Dopa (220.5g) was observed to be better than birds on other diets. Meat to bone ratio and relative organ weights were not affected across treatments. L-Dopa extract, at levels tested, had no detrimental effect on broilers, rather better bird performance and carcass characteristics were observed especially at 0.1% and 0.2% L-Dopa inclusion rates. Therefore, 0.2% inclusion is recommended in diets of broiler chickens for improved performance and carcass characteristics.

Keywords: broilers, carcass characteristics, l-dopa, performance

Procedia PDF Downloads 284

Authors: Obinna K. Ihesiulor, Krishna Shankar, Paul Smith, Alan Fien

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Excessive fretting wear at the taper-trunnion junction (trunnionosis) apparently contributes to the high failure rates of hip implants. Implant wear and corrosion lead to the release of metal particulate debris and subsequent release of metal ions at the taper-trunnion surface. This results in a type of metal poisoning referred to as metallosis. The consequences of metal poisoning include; osteolysis (bone loss), osteoarthritis (pain), aseptic loosening of the prosthesis and revision surgery. Follow up after revision surgery, metal debris particles are commonly found in numerous locations. Background: A stable connection between the femoral ball head (taper) and stem (trunnion) is necessary to prevent relative motions and corrosion at the taper junction. Hence, the importance of component assembly cannot be over-emphasized. Therefore, the aim of this study is to determine the influence of head-stem junction assembly by press fitting and the subsequent disengagement/disassembly on the connection strength between the taper ball head and stem. Methods: CoCr femoral heads were assembled with High stainless hydrogen steel stem (trunnion) by Push-in i.e. press fit; and disengaged by Pull-out test. The strength and stability of the two connections were evaluated by measuring the head pull-out forces according to ISO 7206-10 standards. Findings: The head-stem junction strength linearly increases with assembly forces.

Keywords: wear, modular hip prosthesis, taper head-stem, force assembly and disassembly

Procedia PDF Downloads 372

Authors: Doaa M. Elghannam, Nashwa Khayrat Abousamra, Doaa A. Shahin, Enas F. Goda, Hanan Azzam, Emad Azmy, Manal Salah El-Din

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Background: The pathogenesis of acute myeloid leukemia (AML) involves the cooperation of mutations promoting proliferation/survival and those impairing differentiation. Point mutations of the NRAS gene are the most frequent somatic mutations causing aberrant signal-transduction in acute myeloid leukemia (AML). Aim: The present work was conducted to study the frequency and prognostic significance of NRAS gene mutations (NRASmut) in de novo Egyptian adult AML. Material and methods: Bone marrow specimens from 150 patients with de novo acute myeloid leukemia and controls were analyzed by genomic PCR-SSCP at codons 12, 13 (exon 1), and 61 (exon 2) for NRAS mutations. Results: NRAS gene mutations was found in 19/150 (12.7%) AML cases, represented more frequently in the FAB subtype M4eo (P = 0.028), and at codon 12, 13 (14of 19; 73.7%). Patients with NRASmut had a significant lower peripheral marrow blasts (P = 0.004, P=0.03) and non significant improved clinical outcome than patients without the mutation. Complete remission rate was (63.2% vs 56.5%; p=0.46), resistant disease (15.8% vs 23.6%; p=0.51), three years overall survival (44% vs 42%; P = 0.85) and disease free survival (42.1% vs 38.9%, P = 0.74). Multivariate analysis showed that age was the strongest unfavorable factor for overall survival (relative risk [RR], 1.9; P = .002), followed by cytogenetics (P = .004). FAB types, NRAS mutation, and leukocytosis were less important. Conclusions: NRAS gene mutation frequency and spectrum differ between biologically distinct subtypes of AML but do not significantly influence prognosis and clinical outcome.

Keywords: NRAS Gene, egyptian adult, acute myeloid leukemia, cytogenetics

Procedia PDF Downloads 67

Authors: A. Reum Son, Jin Seon Kwon, Seung Hun Park, Hai Bang Lee, Moon Suk Kim

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These days, stem cell therapy is focused on for promising source of treatment in clinical human disease. As a supporter of stem cells, in situ-forming hydrogels with growth factors and cells appear to be a promising approach in tissue engineering. To examine osteogenic differentiation of hTMSCs which is one of mesenchymal stem cells in vivo in an injectable hydrogel, we use a methoxy polyethylene glycol-polycaprolactone blockcopolymer (MPEG-PCL) solution with osteogenic factors. We synthesized MPEG-PCL hydrogel and measured viscosity to check sol-gel transition. In order to demonstrate osteogenic ability of hTMSCs, we conducted in vitro osteogenesis experiment. Then, to confirm the cell cytotoxicity, we performed WST-1 with hTMSCs and MPEG-PCL. As the result of in vitro experiment, we implanted cell and hydrogel mixture into animal model and checked degree of osteogenesis with histological analysis and amount of expression genes. Through these experimental data, MPEG-PCL hydrogel has sol-gel transition in temperature change and is biocompatible with stem cells. In histological analysis and gene expression, hTMSCs are very good source of osteogenesis with hydrogel and will use it to tissue engineering as important treatment method. hTMSCs could be a good adult stem cell source for usability of isolation and high proliferation. When hTMSCs are used as cell therapy method with in situ-formed hydrogel, they may provide various benefits like a noninvasive alternative for bone tissue engineering applications.

Keywords: injectable hydrogel, stem cell, osteogenic differentiation, tissue engineering

Procedia PDF Downloads 422

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

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Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

Procedia PDF Downloads 290

Authors: Zahra Asgharpour, Eric Renteria, Sebastian De Boodt

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There is a growing trend towards personalization of medical care, as evidenced by the emphasis on outcomes based medicine, the latest developments in CT and MR imaging and personalized treatment in a variety of surgical disciplines. 3D Printing has been introduced and applied in the medical field since 2000. The first applications were in the field of dental implants and custom prosthetics. According to recent publications, 3D printing in the medical field has been used in a wide range of applications which can be organized into several categories including implants, prosthetics, anatomical models and tissue bioprinting. Some of these categories are still in their infancy stage of the concept of proof while others are in application phase such as the design and manufacturing of customized implants and prosthesis. The approach of 3D printing in this category has been successfully used in the health care sector to make both standard and complex implants within a reasonable amount of time. In this study, some of the clinical applications of 3D printing in design and manufacturing of a patient-specific hip implant would be explained. In cases where patients have complex bone geometries or are undergoing a complex revision on hip replacement, the traditional surgical methods are not efficient, and hence these patients require patient-specific approaches. There are major advantages in using this new technology for medical applications, however, in order to get this technology widely accepted in medical device industry, there is a need for gaining more acceptance from the medical device regulatory offices. This is a challenge that is moving onward and will help the technology find its way at the end as an accepted manufacturing method for medical device industry in an international scale. The discussion will conclude with some examples describing the future directions of 3D Medical Printing.

Keywords: CT/MRI, image processing, 3D printing, medical devices, patient specific implants

Procedia PDF Downloads 259

Authors: Rajandeep Kaur, Rajeev Gupta

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Background: Chronic myeloid leukemia (CML) is the most common leukaemia in India. The annual incidence of chronic myeloid leukemia in India was originally reported to be 0.8 to 2.2 per 1,00,000 population. CML is a clonal disorder that is usually easily diagnosed because the leukemic cells of more than 95% of patients have a distinctive cytogenetic abnormality, the Philadelphia chromosome (Ph1). The approval of tyrosine kinase inhibitors (TKIs), which target BCR-ABL1 kinase activity, has significantly reduced the mortality rate associated with chronic myeloid leukemia (CML) and revolutionized treatment. Material and Methods: 80 diagnosed cases of CML were taken. Investigations were done. Bone marrow and molecular studies were also done and with EUTOS, patients were stratified into low and high-risk groups and then treatment with Imatinib was given to all patients and the molecular response was evaluated at 6 months and 12 months follow up with BCR-ABL by RT-PCR quantitative assay. Results: In the study population, out of 80 patients in the study population, 40 were females and 40 were males, with M: F is 1:1. Out of total 80 patients’ maximum patients (54) were in 31-60 years age group. Our study showed a most common symptom of presentation is abdominal discomfort followed by fever. Out of the total 80 patients, 25 (31.3%) patients had high EUTOS scores and 55 (68.8%) patients had low EUTOS scores. On 6 months follow up 36.3% of patients had Complete Molecular Response, 16.3% of patients had Major Molecular Response and 47.5% of patients had No Molecular Response but on 12 months follow up 71.3% of patients had Complete Molecular Response, 16.25% of patients had Major Molecular Response and 12.5% patients had No Molecular Response. Conclusion: In this study, we found a significant correlation between EUTOS score and Molecular response at 6 months and 12 months follow up after Imatinib therapy.

Keywords: chronic myeloid leukemia, European treatment and outcome study score, hematological response, molecular response, tyrosine kinase inhibitor

Procedia PDF Downloads 74

Authors: Chih-Yi Ho, Tzu-Fang Chang, Chih-Chia Huang, Chia-Yen Lee

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In dental clinics, the dentists use the periapical radiography image to assess the effectiveness of endodontic treatment of teeth with chronic apical periodontitis. Periapical radiography images are taken at different times to assess alveolar bone variation before and after the root canal treatment, and furthermore to judge whether the treatment was successful. Current clinical assessment of apical tissue recovery relies only on dentist personal experience. It is difficult to have the same standard and objective interpretations due to the dentist or radiologist personal background and knowledge. If periapical radiography images at the different time could be registered well, the endodontic treatment could be evaluated. In the image registration area, it is necessary to assign representative control points to the transformation model for good performances of registration results. However, detection of representative control points (feature points) on periapical radiography images is generally very difficult. Regardless of which traditional detection methods are practiced, sufficient feature points may not be detected due to the low-contrast characteristics of the x-ray image. Barnard detector is an algorithm for feature point detection based on grayscale value gradients, which can obtain sufficient feature points in the case of gray-scale contrast is not obvious. However, the Barnard detector would detect too many feature points, and they would be too clustered. This study uses the local extrema of clustering feature points and the suppression radius to overcome the problem, and compared different feature point detection methods. In the preliminary result, the feature points could be detected as representative control points by the proposed method.

Keywords: feature detection, Barnard detector, registration, periapical radiography image, endodontic treatment

Procedia PDF Downloads 423

Authors: Ewa Mojs, Katarzyna Wiechec, Maia Kubiak, Wlodzimierz Samborski

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Interactions between mother’s psychological condition and child’s health status are complex and derive from the nature of the mother-child relationship. The aim of the study was to analyze the issue of anxiety amongst mothers of short children in the aspect of growth hormone therapy. The study was based on a group of 101 mothers of originally short-statured children – 70 with growth hormone deficiency (GHD) treated with recombinant human growth hormone (rhGH) and 31 undergoing the diagnostic process, without any treatment. Collected medical data included child's gender, height and weight, chronological age, bone age delay, and rhGH therapy duration. For all children, the height SDS and BMI SDS were calculated. To evaluate anxiety in mothers, the Spielberger State-Trait Anxiety Inventory (STAI) was used. Obtained results revealed low trait anxiety levels, with no statistically significant differences between the groups. State anxiety levels were average when mothers of all children were analyzed together, but when divided into groups, statistical differences appeared. Mothers of children without diagnosis and treatment had significantly higher levels of state anxiety than mothers of children with GHD receiving appropriate therapy. These results show, that the occurrence of growth failure in children is not related to high maternal trait anxiety, but the lack of diagnosis and lack of appropriate treatment generates higher levels of maternal state anxiety than the process of rh GH therapy in the offspring. Commencement of growth hormone therapy induce a substantial reduction of the state anxiety in mothers, and the duration of treatment causes its further decrease.

Keywords: anxiety, development, growth hormone deficiency, motherhood

Procedia PDF Downloads 254

Authors: Shu-Jen Chen, Yi-Chien Wan, Ruey-Chi Wang

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Hydroxyapatite (Ca10(PO4)6(OH)2, HAp) can be applied to the fabrication of bone replacement materials, the composite of dental filling, and the adsorption of biomolecules and dyes. The integration of HAp and magnetic materials would offer several advantages for bio-separation process because the magnetic adsorbents is capable of recovered by applied magnetic field. C-phycocyanin (C-PC) and Allophycocyanin (APC), isolated from Spirulina platensis, can be used in fluorescent labeling probes, health care foods and clinical diagnostic reagents. Although the purification of C-PC and APC are reported by HAp adsorption, the adsorption of C-PC and APC by magnetic HAp composites was not reported yet. Therefore, the fabrication of HAp with magnetic silica nanoparticles for proteins adsorption was investigated in this work. First, the magnetic silica particles were prepared by covering silica layer on Fe3O4 nanoparticles with a reverse micelle method. Then, the Fe3O4@SiO2 nanoparticles were mixed with calcium carbonate to obtain magnetic silica/calcium carbonate composites (Fe3O4@SiO2/CaCO3). The Fe3O4@SiO2/CaCO3 was further reacted with K2HPO4 for preparing the magnetic silica/hydroxyapatite composites (Fe3O4@SiO2/HAp). The adsorption experiments indicated that the adsorption capacity of Fe3O4@SiO2/HAp toward C-PC and APC were highest at pH 6. The adsorption of C-PC and APC by Fe3O4@SiO2/HAp could be correlated by the pseudo-second-order model, indicating chemical adsorption dominating the adsorption process. Furthermore, the adsorption data showed that the adsorption of Fe3O4@SiO2/HAp toward C-PC and APC followed the Langmuir isotherm. The isoelectric points of C-PC and APC were around 5.0. Additionally, the zeta potential data showed the Fe3O4@SiO2/HAp composite was negative charged at pH 6. Accordingly, the adsorption mechanism of Fe3O4@SiO2/HAp toward C-PC and APC should be governed by hydrogen bonding rather than electrostatic interaction. On the other hand, as compared to C-PC, the Fe3O4@SiO2/HAp shows higher adsorption affinity toward APC. Although the Fe3O4@SiO2/HAp cannot recover C-PC and APC from Spirulina platensis homogenate, the Fe3O4@SiO2/HAp can be applied to separate C-PC and APC.

Keywords: hydroxyapatite, magnetic, C-phycocyanin, allophycocyanin

Procedia PDF Downloads 122

Authors: Rupita Ghosh, Ritwik Sarkar, Sumit K. Pal, Soumitra Paul

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The use of Hydroxyapatite bioceramics as restorative implants is widely known. These materials can be manufactured by pressing and sintering route to a particular shape. However machining processes are still a basic requirement to give a near net shape to those implants for ensuring dimensional and geometrical accuracy. In this context, optimising the machining parameters is an important factor to understand the machinability of the materials and to reduce the production cost. In the present study a method has been optimized to produce true particulate drilled composite of Hydroxyapatite Yttrium Phosphate. The phosphates are used in varying ratio for a comparative study on the effect of flexural strength, hardness, machining (drilling) parameters and bioactivity.. The maximum flexural strength and hardness of the composite that could be attained are 46.07 MPa and 1.02 GPa respectively. Drilling is done with a conventional radial drilling machine aided with dynamometer with high speed steel (HSS) and solid carbide (SC) drills. The effect of variation in drilling parameters (cutting speed and feed), cutting tool, batch composition on torque, thrust force and tool wear are studied. It is observed that the thrust force and torque varies greatly with the increase in the speed, feed and yttrium phosphate content in the composite. Significant differences in the thrust and torque are noticed due to the change of the drills as well. Bioactivity study is done in simulated body fluid (SBF) upto 28 days. The growth of the bone like apatite has become denser with the increase in the number of days for all the composition of the composites and it is comparable to that of the pure hydroxyapatite.

Keywords: Bioactivity, Drilling, Hydroxyapatite, Yttrium Phosphate

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Authors: Antarip Bhattacharya, Dhritiman Maitra

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Primary hyperparathyroidism (PHPT) is the most common cause of hypercalcemia due to autonomous production of parathormone (PTH) and the third most common endocrine disorder. Upto 2% of postmenopausal women could have this condition. Primary hyperparathyroidism is characterized by hypercalcemia with a high or insufficiently suppressed level of parathyroid hormone and is caused by a solitary parathyroid adenoma in 85-90% of patients. PHPT may also be caused by parathyroid hyperplasia (involving multiple glands) or parathyroid carcinoma. Associated morbidities and sequelae include decreased bone mineral density, fractures, kidney stones, hypertension, cardiac comorbidities and psychiatric disorder which entail huge costs for treatment. In the year 2021, by virtue of running a Breast and Endocrine Surgery clinic in a Tier 1 city at a tertiary care hospital, the opportunity to be associated with patients of hyperparathyroidism came our way. Here, we shall describe the spectrum of clinical presentations and customisation of treatment for parathyroid diseases with reference to the above patients. A retrospective analysis of the data of all patients presenting with symptoms of parathyroid diseases was made and classified according to the cause. 13 patients had presented with symptoms of hyperparathyroidism and each case presented with unique symptoms and necessitated detailed evaluation. The treatment or surgery offered to each patient was tailored to his/her individual disease and led to favourable outcomes. Diseases affecting parathyroid are not as rare as we believe. Each case merits detailed clinical evaluation, investigations and tailoring of suitable treatment with regard to medical management and extent of surgery. Intra-operative frozen section/iOPTH monitoring are really useful adjuncts for intra-operative decision making.

Keywords: hyperparathyroidism, parathyroid adenoma, parathyroid surgery, PTH

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Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler

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Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.

Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome

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Authors: Marija Zulić, Vanda Hájková, Nina Brkić–Jovanović, Srećko Potić, Sanja Tomić

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The name cerebral palsy comes from the word cerebrum, which means the brain and the word palsy, which means seizure, and essentially refers to the movement disorder. In the clinical picture of cerebral palsy, basic neuromotor disorders are associated with other various disorders: behavioural, intellectual, speech, sensory, epileptic seizures, and bone and joint deformities. Motor speech disorders are among the most common difficulties present in people with cerebral palsy. Social participation represents an interaction between an individual and their social environment. Quality of social participation of the students with cerebral palsy at school is an important indicator of their successful participation in adulthood. One of the most important skills for the undisturbed social participation is ability of good communication. The aim of the study was to determine relation between social participation of students with cerebral palsy and presence of their speech impairment in primary schools in the Czech Republic. The study was performed in the Czech Republic in mainstream schools and schools established for the pupils with special education needs. We analysed 75 children with cerebral palsy aged between six and twelve years attending up to sixth grade by using the first and the third part of the school function assessment questionnaire as the main instrument. The other instrument we used in the research is the Gross motor function classification system–five–level classification system, which measures degree of motor functions of children and youth with cerebral palsy. Funding for this study was provided by the Grant Agency of Charles University in Prague.

Keywords: cerebral palsy, social participation, speech disorders, The Czech Republic, the school function assessment

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Authors: Recep Duzsoz, Ozlem Gormez, Umit Memis, Selma Demer, Hikmet Orhan

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Stylohyoid ligament is a connective tissue extending from apex of the styloid process to small horn of the hyoid bone. The normal length of styloid process ranges from 20 to 30 mm and measurements more than 30 mm is named stylohyoid ligament calcification (SLC). Fluorosis is a health problem that arises in individuals who intake large amounts of fluor long periods of time. The aim of this study was to investigate the effects of fluorosis on SLC. This study has been conducted on 100 patients who had SLC detected on panoramic radiograph. The study group was consisted of 50 patients with dental fluorosis and control group was consisted of 50 patients without dental fluorosis. Length and thickness of SLC were measured and the type of SLC was determined on panoramic radiographs. There was no statistically significant differences between the study and control group for SLC length, thickness and type. The thickness of left and right SLC of severe dental fluorosis group was statistically significant higher than moderate dental fluorosis group (p < 0,05). Cervicopharyngeal trauma, tonsillectomy, endocrine disease in menopause, persistent mesenchymal tissue, mechanical stress have reported as etiology of SLC in the literature and studies are still ongoing. It was reported that fluorosis as a factor on calcification of some ligaments in body (posterior longitudunal ligament, ligamentum flavum and transverse atlantal ligament) previously but relationship between fluorosis with SLC was not investigated. Our study is unique because it is the first study on SLC thickness measurements on panoramic radiographs and the relationship between fluorosis and SLC to our knowledge. According to the obtained results, it is thought that fluorosis may have an effect on SLC in thickness due to the relationship between dental fluorosis severity with SLC thickness and this study will contribute to the progress of the future studies.

Keywords: calcification, fluorosis, ligament, stylohyoid

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Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu

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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.

Keywords: inflammasome, inflammation, innate immunity, pyroptosis

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Authors: Naeim Akbari Shahkhosravi, Reza Kakavand, Helen M. S. Davies, Amin Komeili

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Equine locomotion and performance are significantly affected by hoof health. One of the most critical diseases of the hoof is laminitis, which can lead to horse lameness in a severe condition. This disease exhibits the mechanical properties degradation of the laminar junction tissue within the hoof. Therefore, it is essential to investigate the biomechanics of the hoof, focusing specifically on excessive and cumulatively accumulated stresses within the laminar junction tissue. For this aim, the current study generated a novel equine hoof Finite Element (FE) model under dynamic physiological loading conditions and employing a hyperelastic material model. Associated tissues of the equine hoof were segmented from computed tomography scans of an equine forelimb, including the navicular bone, third phalanx, sole, frog, laminar junction, digital cushion, and medial- dorsal- lateral wall areas. The inner tissues were connected based on the hoof anatomy, and the hoof was under a dynamic loading over cyclic strides at the trot. The strain distribution on the hoof wall of the model was compared with the published in vivo strain measurements to validate the model. Then the validated model was used to study the development of laminitis. The ultimate stress tolerated by the laminar junction before rupture was considered as a stress threshold. The tissue damage was simulated through iterative reduction of the tissue’s mechanical properties in the presence of excessive maximum principal stresses. The findings of this investigation revealed how damage initiates from the medial and lateral sides of the tissue and propagates through the hoof dorsal area.

Keywords: horse hoof, laminitis, finite element model, continuous damage

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Authors: Pranee Suecharoen, Jaturat Kanpittaya

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Background: The incidence of adverse reactions to iodinated contrast media has risen. The dearth of reports on reactions to the administration of iso- and low-osmolar contrast media should be addressed. We, therefore, studied the profile of adverse reactions to iodinated contrast media; viz., (a) the body systems affected (b) causality, (c) severity, and (d) preventability. Objective: To study adverse reactions (causes and severity) to iodinated contrast media at Srinagarind Hospital. Method: Between March and July, 2015, 1,101 patients from the Department of Radiology were observed and interviewed for the occurrence of adverse reactions. The patients were classified per Naranjo’s algorithm and through use of an adverse reactions questionnaire. Results: A total of 105 cases (9.5%) reported adverse reactions (57% male; 43% female); among whom 2% were iso-osmolar vs. 98% low-osmolar. Diagnoses included hepatoma and cholangiocarcinoma (24.8%), colorectal cancer (9.5%), breast cancer (5.7%), cervical cancer (3.8%), lung cancer (2.9%), bone cancer (1.9%), and others (51.5%). Underlying diseases included hypertension and diabetes mellitus type 2. Mild, moderate, and severe adverse reactions accounted for 92, 5 and 3%, respectively. The respective groups of escalating symptoms included (a) mild urticaria, itching, rash, nausea, vomiting, dizziness, and headache; (b) moderate hypertension, hypotension, dyspnea, tachycardia and bronchospasm; and (c) severe laryngeal edema, profound hypotension, and convulsions. All reactions could be anticipated per Naranjo’s algorithm. Conclusion: Mild to moderate adverse reactions to low-osmolar contrast media were most common and these occurred immediately after administration. For patient safety and better outcomes, improving the identification of patients likely to have an adverse reaction is essential.

Keywords: adverse reactions, contrast media, computed tomography, iodinated contrast agents

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Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

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The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

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Authors: Morteza Elsa, Amirhossein Moghanian

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The major aim of this study was to evaluate the effect of CaO content on in vitro hydroxyapatite formation, MC3T3 cells cytotoxicity and proliferation as well as antibacterial efficiency of sol-gel derived SiO₂–CaO–P₂O₅ ternary system. For this purpose, first two grades of bioactive glass (BG); BG-58s (mol%: 60%SiO₂–36%CaO–4%P₂O₅) and BG-68s (mol%: 70%SiO₂–26%CaO–4%P₂O₅)) were synthesized by sol-gel method. Second, the effect of CaO content in their composition on in vitro bioactivity was investigated by soaking the BG-58s and BG-68s powders in simulated body fluid (SBF) for time periods up to 14 days and followed by characterization inductively coupled plasma atomic emission spectrometry (ICP-AES), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. Additionally, live/dead staining, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity assays were conducted respectively, as qualitatively and quantitatively assess for cell viability, proliferation and differentiations of MC3T3 cells in presence of 58s and 68s BGs. Results showed that BG-58s with higher CaO content showed higher in vitro bioactivity with respect to BG-68s. Moreover, the dissolution rate was inversely proportional to oxygen density of the BG. Live/dead assay revealed that both 58s and 68s increased the mean number live cells which were in good accordance with MTT assay. Furthermore, BG-58s showed more potential antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Taken together, BG-58s with enhanced MC3T3 cells proliferation and ALP activity, acceptable bioactivity and significant high antibacterial effect against MRSA bacteria is suggested as a suitable candidate in order to further functionalizing for delivery of therapeutic ions and growth factors in bone tissue engineering.

Keywords: antibacterial, bioactive glass, hydroxyapatite, proliferation, sol-gel processes

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Authors: Jingling Li, Yi Zhang, Wei Liang, Tao Cui, Jun Li

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Compared with terrestrial network, the traffic of spatial information network has both self-similarity and short correlation characteristics. By studying its traffic prediction method, the resource utilization of spatial information network can be improved, and the method can provide an important basis for traffic planning of a spatial information network. In this paper, considering the accuracy and complexity of the algorithm, the spatial information network traffic is decomposed into approximate component with long correlation and detail component with short correlation, and a time series hybrid prediction model based on wavelet decomposition is proposed to predict the spatial network traffic. Firstly, the original traffic data are decomposed to approximate components and detail components by using wavelet decomposition algorithm. According to the autocorrelation and partial correlation smearing and truncation characteristics of each component, the corresponding model (AR/MA/ARMA) of each detail component can be directly established, while the type of approximate component modeling can be established by ARIMA model after smoothing. Finally, the prediction results of the multiple models are fitted to obtain the prediction results of the original data. The method not only considers the self-similarity of a spatial information network, but also takes into account the short correlation caused by network burst information, which is verified by using the measured data of a certain back bone network released by the MAWI working group in 2018. Compared with the typical time series model, the predicted data of hybrid model is closer to the real traffic data and has a smaller relative root means square error, which is more suitable for a spatial information network.

Keywords: spatial information network, traffic prediction, wavelet decomposition, time series model

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Authors: Wei Wen Yang, Emily Chia Yu Su

Abstract:

Dengue fever is a mosquito-borne tropical infectious disease caused by the dengue virus. After infection, symptoms usually start from three to fourteen days. Dengue virus may cause a high fever and at least two of the following symptoms, severe headache, severe eye pain, joint pains, muscle or bone pain, vomiting, feature skin rash, and mild bleeding manifestation. In addition, recovery will take at least two to seven days. Dengue fever has rapidly spread in tropical and subtropical areas in recent years. Several phenomena around the world such as global warming, urbanization, and international travel are the main reasons in boosting the spread of dengue. In Taiwan, epidemics occur annually, especially during summer and fall seasons. On the other side, Singapore government also has announced the amounts number of dengue cases spreading in Singapore. As the serious epidemic of dengue fever outbreaks in Taiwan and Singapore, countries around the Asia-Pacific region are becoming high risks of susceptible to the outbreaks and local hub of spreading the virus. To improve public safety and public health issues, firstly, we are going to use Microsoft Excel and SAS EG to do data preprocessing. Secondly, using support vector machines and decision trees builds predict model, and analyzes the infectious cases between Taiwan and Singapore. By comparing different factors causing vector mosquito from model classification and regression, we can find similar spreading patterns where the disease occurred most frequently. The result can provide sufficient information to predict the future dengue infection outbreaks and control the diffusion of dengue fever among countries.

Keywords: dengue fever, Taiwan, Singapore, Aedes aegypti

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Authors: Gina Leisching, Ray-Dean Pietersen, Carel Van Heerden, Paul Van Helden, Ian Wiid, Bienyameen Baker

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The distinguishing factors that characterize the host response to infection with virulent Mycobacterium tuberculosis (M.tb) are largely confounding. We present an infection study with two genetically closely related M.tb strains that have vastly different pathogenic characteristics. The early host response to infection with these detergent-free cultured strains was analyzed through RNAseq in an attempt to provide information on the subtleties which may ultimately contribute to the virulent phenotype. Murine bone marrow-derived macrophages (BMDMs) were infected with either a hyper- (R5527) or hypovirulent (R1507) Beijing M. tuberculosis clinical isolate. RNAseq revealed 69 differentially expressed host genes in BMDMs during comparison of these two transcriptomes. Pathway analysis revealed activation of the stress-induced and growth inhibitory Gadd45 signaling pathway in hypervirulent infected BMDMs. Upstream regulators of interferon activation such as and IRF3 and IRF7 were predicted to be upregulated in hypovirulent-infected BMDMs. Additional analysis of the host immune response through ELISA and qPCR included the use of human THP-1 macrophages where a robust proinflammatory response was observed after infection with the hypervirulent strain. RNAseq revealed two early-response genes (IER3 and SAA3) and two host-defence genes (OASL1 and SLPI) that were significantly upregulated by the hypervirulent strain. The role of these genes under M.tb infection conditions are largely unknown but here we provide validation of their presence with use of qPCR and Western blot. Further analysis into their biological role under infection with virulent M.tb is required.

Keywords: host-response, Mycobacterium tuberculosis, RNAseq, virulence

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Authors: Alireza Arab, Morteza Elsa, Amirhossein Moghanian

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In this study, experiments were carried out to achieve a promising multifunctional and modified silicate based bioactive glass (BG). The main aim of the study was investigating the effect of lithium (Li) and magnesium (Mg) substitution, on in vitro bioactivity of substituted-58S BG. Moreover, it is noteworthy to state that modified BGs were synthesized in 60SiO₂–(36-x)CaO–4P₂O₅–(x)Li₂O and 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO (where x = 0, 5, 10 mol.%) quaternary systems, by sol-gel method. Their performance was investigated through different aspects such as biocompatibility, antibacterial activity as well as their effect on alkaline phosphatase (ALP) activity, and proliferation of MC3T3 cells. The antibacterial efficiency was evaluated against methicillin-resistant Staphylococcus aureus bacteria. To do so, CaO was substituted with Li₂O and MgO up to 10 mol % in 58S-BGs and then samples were immersed in simulated body fluid up to 14 days and then, characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry, and scanning electron microscopy. Results indicated that this modification led to a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium revealed further pronounced effect. The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and ALP analysis illustrated that substitutions of both Li₂O and MgO, up to 5 mol %, had increasing effect on biocompatibility and stimulating proliferation of the pre-osteoblast MC3T3 cells in comparison to the control specimen. Regarding to bactericidal efficiency, the substitution of either Li or Mg for Ca in the 58s BG composition led to statistically significant difference in antibacterial behaviors of substituted-BGs. Meanwhile, the sample containing 5 mol % CaO/Li₂O substitution (BG-5L) was selected as a multifunctional biomaterial in bone repair/regeneration due to the improved biocompatibility, enhanced ALP activity and antibacterial efficiency among all of the synthesized L-BGs and M-BGs.

Keywords: alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes

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Authors: Gi-Hoon Yang, JongHan Ha, MyungGu Yeo, JaeYoon Lee, SeungHyun Ahn, Hyeongjin Lee, HoJun Jeon, YongBok Kim, Minseong Kim, GeunHyung Kim

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Tissue engineering scaffolds must be biocompatible and biodegradable, provide adequate mechanical strength and cell attachment site for proliferation and differentiation. Furthermore, the scaffold morphology (such as pore size, porosity and pore interconnectivity) plays an important role. The electrospinning process has been widely used to fabricate micro/nano-sized fibres. Electrospinning allows for the fabrication of non-woven meshes containing micro- to nano-sized fibers providing high surface-to-volume area for cell attachment. Due to its advantageous characteristics, electrospinning is a useful method for skin, cartilage, bone, and nerve regeneration. In this study, we fabricated PCL scaffolds (SP) consisting of spiral-like struts using 3D melt-plotting system and micro/nanofibers using direct electrospinning writing. By altering the conditions of the conventional melt-plotting method, spiral-like struts were generated. Then, micro/nanofibers were deposited selectively. The control scaffold composed of perpendicular PCL struts was fabricated using the conventional melt-plotting method to compare the cellular activities. The effect on the attached cells (osteoblast-like cells (MG63)) was evaluated depending on the bending instability of the struts. The SP scaffolds showed enhanced biological properties such as initial cell attachment, proliferation and osteogenic differentiation. These results suggest that the SP scaffolds has potential as a bioengineered substitute for soft and hard tissue regeneration.

Keywords: cell attachment, electrospinning, mechanical strength, melt-plotting

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Authors: Garg Ramneesh, Uppal Sanjeev, Mittal Rajinder, Shah Sheerin, Jain Vikas, Singla Bhupinder

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Introduction: Despite advances in trauma care, a classification system for soft tissue injuries of the face still needs to be objectively defined. Aim: To develop a classification system for soft tissue injuries of the face; that is objective, easy to remember, reproducible, universally applicable, aids in surgical management and helps to develop a structured data that can be used for future use. Material and Methods: This classification system includes those patients that need surgical management of facial injuries. Associated underlying bony fractures have been intentionally excluded. Depending upon the severity of soft tissue injury, these can be graded from 0 to IV (O-Abrasions, I-lacerations, II-Avulsion injuries with no skin loss, III-Avulsion injuries with skin loss that would need graft or flap cover, and IV-complex injuries). Anatomically, the face has been divided into three zones (Zone 1/2/3), as per aesthetic subunits. Zone 1e stands for injury of eyebrows; Zones 2 a/b/c stand for nose, upper eyelid and lower eyelid respectively; Zones 3 a/b/c stand for upper lip, lower lip and cheek respectively. Suffices R and L stand for right or left involved side, B for presence of foreign body like glass or pellets, C for extensive contamination and D for depth which can be graded as D 1/2/3 if depth is still fat, muscle or bone respectively. I is for damage to facial nerve or parotid duct. Results and conclusions: This classification system is easy to remember, clinically applicable and would help in standardization of surgical management of soft tissue injuries of face. Certain inherent limitations of this classification system are inability to classify sutured wounds, hematomas and injuries along or against Langer’s lines.

Keywords: soft tissue injuries, face, avulsion, classification

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Authors: Somaiya Mateen, Shagufta Moin, Mohammad Owais, Abdul Khan, Atif Zafar

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In rheumatoid arthritis (RA), impaired oxidative metabolism and imbalance between pro-and anti-inflammatory cytokines are responsible for causing inflammation and the degradation of cartilage and bone. The present study was done to evaluate the level and hence the role of reactive oxygen species (ROS) and inflammatory cytokines in the pathogenesis of RA. The present study was performed in the blood of 80 RA patients and 55 age and sex-matched healthy controls. The level of ROS (in 5% hematocrit) and the plasma level of pro-inflammatory cytokines [TNF-α, interleukin-6 (IL-6), IL-22] and anti-inflammatory cytokines (IL-4 and IL-5) were monitored in healthy subjects and RA patients. For evaluating the role of rheumatoid factor (RF) in the pathogenesis of RA, patients were sub-divided on the basis of presence or absence of RF. Reactive species and inflammatory cytokines were correlated with disease activity measure-Disease Activity Score for 28 joints (DAS28). The level of ROS, TNF-α, IL-6 and IL-22 were found to be significantly higher in RA patients as compared to the healthy controls, with the increase being more significant in patients positive for rheumatoid factor and those having high disease severity. On the other hand, a significant decrease in the level of IL-4 and IL-10 were observed in RA patients compared with healthy controls, with the decrease being more prominent in severe cases of RA. Higher ROS (indicative of impaired anti-oxidant defence system) and pro-inflammatory cytokines level in RA patients may lead to the damage of biomolecules which in turn contributes to tissue damage and hence to the development of more severe RA. The imbalance between pro-and anti-inflammatory cytokines may lead to the development of multi-system immune complications. ROS and inflammatory cytokines may also serve as a potential biomarker for assessing the disease severity.

Keywords: rheumatoid arthritis, reactive oxygen species, pro-inflammatory cytokines, anti-inflammatory cytokines

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Authors: H. Hadjeris, R. B. Ghoul, Lekhlaf, M. Nebbal

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Introduction: Adenoid cystic carcinomas of the lacrimal gland or orbital cylindroma constitute the second cause of epithelial tumors of this gland. It is a malignant tumor usually developed at the expense of the salivary glands; its orbital location is exceptional. It is a rare clinical entity, formidable by its malignancy and local aggressiveness; the recurrence rate is high. Materials and methods: Clinical case: 63 years old woman who presents with irreducible no pulsatile painful left exophthalmos with inflammatory chemosis and a decrease in visual acuity with a moderate intracranial hypertension syndrome that has been evolving for 03 months. Antecedent; a biopsy of the tumor was made; the histological examination was in favor of an adenoid cystic carcinoma of the lacrimal gland. Lesion assessment: computed tomography and brain MRI: show an intra and extra-conical mass; with sinus (ethmoido-frontal) and cerebral (left frontal) extension strongly enhanced after injection of contrast product surrounded by edema around the lesion, associated with left frontal bone lysis extension assessment: unremarkable treatment: Patient operated by left frontotemporal approach, a total exenteration was performed with macroscopically complete excision of the frontal lesion and wide frontal craniectomy with craniofacial reconstruction, followed by complementary radiotherapy. Results: The patient was seen again after 3 months in consultation; she does not present any signs in favor of a recurrence. Conclusion: Adenoid cystic carcinomas of the lacrimal gland are rare malignant tumors; they are very infiltrating and invasive. The prognosis is strongly linked to the treatment time.

Keywords: adenoid cystic, lacrimal gland, orbital location, fronto-temporal approac

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Authors: Lay Poh Tan, Chor Yong Tay, Haiyang Yu

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Micro-contact printing is a form of soft lithography that uses the relief patterns on a master polydimethylsiloxane (PDMS) stamp to form patterns of self-assembled monolayers (SAMs) of ink on the surface of a substrate through conformal contact technique. Here, we adopt this method to print proteins of different dimensions on our biodegradable polymer substrates. We started off with printing 20-500 μm scale lanes of fibronectin to engineer the shape of bone marrow derived human mesenchymal stem cell (hMSCs). After 8 hours of culture, the hMSCs adopted elongated shapes, and upon analysis of the gene expressions, genes commonly associated with myogenesis (GATA-4, MyoD1, cTnT and β-MHC) and neurogenesis (NeuroD, Nestin, GFAP, and MAP2) were up-regulated but gene expression associated to osteogenesis (ALPL, RUNX2, and SPARC) were either down modulated or remained at the nominal level. This is the first evidence that cellular morphology control via micropatterning could be used to modulate stem cell fate without external biochemical stimuli. We further our studies to modulate the focal adhesion (FA) instead of the macro shape of cells. Micro-contact printed islands of different smaller dimensions were investigated. We successfully regulated the FAs into dense FAs and elongated FAs by micropatterning. Additionally, the combined effects of hard (40.4 kPa), and intermediate (10.6 kPa) PA gel and FAs patterning on hMSCs differentiation were studied. Results showed that FA and matrix compliance plays an important role in hMSCs differentiation, and there is a cross-talk between different physical stimulants and the significance of these stimuli can only be realized if they are combined at the optimum level.

Keywords: micro-contact printing, polymer substrate, cell-material interaction, stem cell differentiation

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Authors: Matej Patzelt, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Vladimir Musil, Jitka Riedlova, Viktor Sykora, Jana Mrzilkova, Petr Zach

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Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to create a new fixation method for soft tissue imaging in micro-CT. Methodology: We used organs of 18 mice - heart, lungs, kidneys, liver and brain, which we fixated in ethanol after meticulous preparation. We fixated organs in different concentrations of ethanol and for different period of time. We used three types of ethanol concentration - 97%, 50% and ascending ethanol concentration (25%, 50%, 75%, 97% each for 12 hours). Fixated organs were scanned after 72 hours, 168 hours and 336 hours period of fixation. We scanned all specimens in micro-CT MARS (Medipix All Resolution System). Results: Ethanol method provided contrast enhancement in all studied organs in all used types of fixation. Fixation in 97% ethanol provided very fast fixation and the contrast among the tissues was visible already after 72 hours of fixation. Fixation for the period of 168 and 336 hours gave better details, especially in lung tissue, where alveoli were visualized. On the other hand, this type of fixation caused organs to petrify. Fixation in 50% ethanol provided best results in 336 hours fixation, details were visualized better than in 97% ethanol and samples were not as hard as in fixation in 97% ethanol. Best results were obtained in fixation in ascending ethanol concentration. All organs were visualized in great details, best-visualized organ was heart, where trabeculae and valves were visible. In this type of fixation, organs stayed soft for whole time. Conclusion: New ethanol method is a great option for soft tissue fixation as well as the method for enhancing contrast among tissues in organs. The best results were obtained with fixation of the organs in ascending ethanol concentration, the best visualized organ was the heart.

Keywords: x-ray imaging, small animals, ethanol, ex-vivo

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