Search results for: frozen tissue
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1679

Search results for: frozen tissue

1079 The Angiogenic Activity of α-Mangostin in the Development of Zebrafish Embryo In Vivo

Authors: Titis Indah Adi Rahayu

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Angiogenesis is the process of generating new capillary from pre-existing blood vessels. VEGFA is a major regulator in angiogenesis that binds and activates two tyrosine kinase receptors, VEGFR1 (Flt-1) and VEGFR2 (Flk-1/KDR) which regulate pathological and physiological angiogenesis. Disruption of VEGFA and VEGFR2 regulation lead to many diseases. The study of α-Mangostin (derivate of xanthone) as anti-oxidant and anti inflammation has been explored recently and both of them have relation to vasculature however the effect of α-Mangostin in blood vessel formation in healthy tissue in vivo has not been studied. Zebrafish is a powerful model in studying angiogenesis and shared many advantages that is a viable whole animal model for screening small molecules that affect blood vessel formation. Therefore the aim of this study is to evaluate angiogenic activity of α-Mangostin in healthy tissue in vivo in zebrafish embryo in relation of patterning blood vessel. Blood vessel patterning is highly characteristic in the developing of zebrafish embryo and the subintestinal vessel (SIV) can be stained and visualized microscopically as a primary screen for α-Mangostin that effect angiogenesis. The zebrafish embryos are divided into 2 groups. Group one consists of the zebrafish embryos at 1 dpf for 4 days which are tested to α-Mangostin in several concentration 2 µM, 4 µM, 6 µM, 8 µM and 10 µM whereas in group two the zebrafish larva at 4 dpf are exposed to α-Mangostin 1,75 µM, 2,3 µM, 2,9 µM, 3,8 µM dan 5 µM for 2 days. DMSO is served as a control for each group. The level expression of vegfa and vegfr2 are observed quantitatively using real time q-PCR and patterning of SIV are then analized via alkaline phospatase staining. Result shows that the level expression of vegfa and vegfr2 is repressed quantitatively as shown in real time q-PCR in the group of 1-4 days of α-Mangostin exposure where it is increased in the group of 4-6 days of α-Mangostin exposure. The result is then compared to alkaline phospatase staining of SIV using stereo microscope. It indicates that α-Mangostin does not disturb the patterning of SIV formation in zebrafish.

Keywords: angiogenesis, Danio rerio, α-Mangostin, SIV, vegfa, vegfr2

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1078 An Unexpected Hand Injury with Pluridigital Fractures Due to Premature Explosion of a Ramadan Cannon

Authors: Hakan Akgul

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Purpose: The use of firecrackers (i.e., Ramadan Cannon) during the month of Ramadan is a traditional way of indicating that the fasting period is over in Muslim countries. Here, we report the rehabilitation of a case of hand injury with pluridigital fractures due to premature explosion of a Ramadan cannon. Materials and Methods: A 48-year old man admitted to the Emergency Department due to left hand injury as a result of a premature explosion of a Ramadan cannon. The patient was immediately taken to operation room because of the multiple fractures, tendon loss, and soft tissue loss in the left hand. Range of motion (ROM) of joints was measured with goniometer, pain and oedema were measured and splinting was performed. Results: Rehabilitation team took over the patient at postoperative 9th week. During the 3 month rehabilitation, range of motion increased, oedema was taken under control, pain was reduced, the colour of the skin turned to the normal tone. According to the visual analog scale (VAS), pain decreased from 9 to 4. Oedema, around the metacarpofalangeal (MCP) joints, decreased from 27,5 cm to 23,5 cm. Total active range of motion of the wrist increased from 5 degrees to 50 degrees.Total active range of motion of supination and pronation increased from 55 degrees to 70 degrees. Discussion: The rehabilitation of multiple hand injury is quite difficult. Different aspects of trauma should be taken into consideration when rehabilitation is planned. Factors such as waiting for the bone union, wound healing, and use of external fixators may delay rehabilitation process. Joint mobilization, massage for reducing oedema and preventing scar tissue, exercise within the range of motion are efficient measures. Poor patient compliance to treatment may lead to poor outcome. First of all, oedema and scar formation must be taken under control. Removing fixators should not be delayed depending on the bone union, and exercise within the range of motion should be started.

Keywords: explosion, fracture, hand, injury

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1077 Comparative Study of Active Release Technique and Myofascial Release Technique in Patients with Upper Trapezius Spasm

Authors: Harihara Prakash Ramanathan, Daksha Mishra, Ankita Dhaduk

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Relevance: This qualitative study will educate the clinician in putting into practice the advanced method of movement science in restoring the function. Purpose: The purpose of this study is to compare the effectiveness of Active Release Technique and myofascial release technique on range of motion, neck function and pain in patients with upper trapezius spasm. Methods/Analysis: The study was approved by the institutional Human Research and Ethics committee. This study included sixty patients of age group between 20 to 55 years with upper trapezius spasm. Patients were randomly divided into two groups receiving Active Release Technique (Group A) and Myofascial Release Technique (Group B). The patients were treated for 1 week and three outcome measures ROM, pain and functional level were measured using Goniometer, Visual analog scale(VAS), Neck disability Index Questionnaire(NDI) respectively. Paired Sample 't' test was used to compare the differences of pre and post intervention values of Cervical Range of motion, Neck disability Index, Visual analog scale of Group A and Group B. Independent't' test was used to compare the differences between two groups in terms of improvement in cervical range of motion, decrease in visual analogue scale(VAS), decrease in Neck disability index score. Results: Both the groups showed statistically significant improvements in cervical ROM, reduction in pain and in NDI scores. However, mean change in Cervical flexion, cervical extension, right side flexion, left side flexion, right side rotation, left side rotation, pain, neck disability level showed statistically significant improvement (P < 0. 05)) in the patients who received Active Release Technique as compared to Myofascial release technique. Discussion and conclusions: In present study, the average improvement immediately post intervention is significantly greater as compared to before treatment but there is even more improvement after seven sessions as compared to single session. Hence, this proves that several sessions of Manual techniques are necessary to produce clinically relevant results. Active release technique help to reduce the pain threshold by removing adhesion and promote normal tissue extensibility. The act of tensioning and compressing the affected tissue both with digital contact and through the active movement performed by the patient can be a plausible mechanism for tissue healing in this study. This study concluded that both Active Release Technique (ART) and Myofascial release technique (MFR) are equally effective in managing upper trapezius muscle spasm, but more improvement can be achieved by Active Release Technique (ART). Impact and Implications: Active Release Technique can be adopted as mainstay of treatment approach in treating trapezius spasm for faster relief and improving the functional status.

Keywords: trapezius spasm, myofascial release, active release technique, pain

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1076 Nutritive Value of Three-Stage Olive Cake (Olea europaea L.) for Growing Rabbit

Authors: Zahia Dorbane, Si Ammar Kadi, Dalila Boudouma, Thierry Gidenne

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In rabbits feeding, minimum fibre intake is essential to avoid digestive disorders. However, this concentration of fibre is not easy to obtain when formulating feeds, without reduction of nutritional value. Three stage olive cake, the residual material after oil extraction by centrifugation, including pulp and stones, can be used as a fibre source in rabbit diet. The incorporation of olive cake can allow a better balance between different fibre fractions and reduce health disorder. However, for practical use of any raw material, it is necessary to know its chemical and nutritive value. The aim of this study was to assess the nutritive value of three-stage olive cake (TSOC) for growing rabbits. Thus, 36 rabbits weaned at 35 days (702.8 ± 28.5) were divided into three groups of 12 receiving one of the following diets: control with 0% of TSOC, TSOC10 (10% of TSOC) and TSOC20 (20% TSOC). The rabbits were individually housed in digestibility cages and received ad libitum one of the three diets, fresh and clean water was provided ad libitum. After an adaptation period of 7d, feces were collected for 4d. Collected feces were frozen and stored for further analysis. The chemical composition of TSOC shows that it is a rich fiber raw material since it contains (%DM): 6% of CP; 7.4% of EE; 78.7% of NDF; 55.4% of ADF and 24.3% of ADL. The inclusion of TSOC at 20% of basal diet reduced the digestibility coefficient of organic matter, crude protein and NDF from 67.8 to 55.3%, 80.4 to 75.3% and from 31.5 to 18.4% (p < 0.001) respectively. The digestible energy and digestible protein content of the three-stage olive cake estimated by regression was 2.94 ± 0.52MJ DE/kg DM and 22.4 ± 6 g DP/kg DM respectively. In conclusion, based on the results of the present experiment, the three-stage olive cake can be used as a fibre source for rabbit.

Keywords: digestibility, nutritive value, olive cake, rabbit

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1075 Finite Element Modeling of Two-Phase Microstructure during Metal Cutting

Authors: Junior Nomani

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This paper presents a novel approach to modelling the metal cutting of duplex stainless steels, a two-phase alloy regarded as a difficult-to-machine material. Calculation and control of shear strain and stresses during cutting are essential to achievement of ideal cutting conditions. Too low or too high leads to higher required cutting force or excessive heat generation causing premature tool wear failure. A 2D finite element cutting model was created based on electron backscatter diffraction (EBSD) data imagery of duplex microstructure. A mesh was generated using ‘object-oriented’ software OOF2 version V2.1.11, converting microstructural images to quadrilateral elements. A virtual workpiece was created on ABAQUS modelling software where a rigid body toolpiece advanced towards workpiece simulating chip formation, generating serrated edge chip formation cutting. Model results found calculated stress strain contour plots correlated well with similar finite element models tied with austenite stainless steel alloys. Virtual chip form profile is also similar compared experimental frozen machining chip samples. The output model data provides new insight description of strain behavior of two phase material on how it transitions from workpiece into the chip.

Keywords: Duplex stainless steel, ABAQUS, OOF2, Chip formation

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1074 Comparison of Conjunctival Autograft versus Amniotic Membrane Transplantation for Pterygium Surgery

Authors: Luksanaporn Krungkraipetch

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Currently, surgery is the only known effective treatment for pterygium. In certain groups, the probability of recurrence after basic sclera excision is very significant. Tissue grafting is substantially more time-consuming and challenging than keeping the sclera uncovered, but it reduces the chance of recurrence. Conjunctival autograft surgery is older than amniotic membrane graft surgery. The purpose of this study was to compare pterygium surgery with conjunctival autograft against an amniotic membrane transplant. In the study, a randomized controlled trial was used. Four cases were ruled out (two for failing to meet inclusion criteria and the other for refusing to participate). Group I (n = 40) received the intervention, whereas Group II (n = 40) served as the control. Both descriptive and inferential statistical approaches were used, including data analysis and data analysis statistics. The descriptive statistics analysis covered basic pterygium surgery information as well as the risk of recurrent pterygium. As an inferential statistic, the chi-square was used. A p-value of 0.05 is statistically significant. The findings of this investigation were the majority of patients in Group I were female (70.0%), aged 41–60 years, had no underlying disease (95.0%), and had nasal pterygium (97.5%). The majority of Group II patients were female (60.0%), aged 41–60 years, had no underlying disease (97.5%) and had nasal pterygium (97.5%). Group I had no recurrence of pterygium after surgery, but Group II had a 7.5% recurrence rate. Typically, the recurrence time is twelve months. The majority of pterygium recurrences occur in females (83.3%), between the ages of 41 and 60 (66.7%), with no underlying disease. The recurrence period is typically six months (60%) and a nasal pterygium site (83.3%). Pterygium recurrence after surgery is associated with nasal location (p =.002). 16.7% of pterygium surgeries result in complications; one woman with nasal pterygium underwent autograft surgery six months later. The presence of granulation tissue at the surgical site is a mild complication. A pterygium surgery recurrence rate comparison of conjunctival autograft and amniotic membrane transplantation revealed that conjunctival autograft had a higher recurrence rate than amniotic membrane transplantation (p =.013).

Keywords: pterygium, pterygium surgery, conjunctival autograft, amniotic membrane transplantation

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1073 Non-Invasive Pre-Implantation Genetic Assessment Using NGS in IVF Clinical Routine

Authors: Katalin Gombos, Bence Gálik, Krisztina Ildikó Kalács, Krisztina Gödöny, Ákos Várnagy, József Bódis, Attila Gyenesei, Gábor L. Kovács

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Although non-invasive pre-implantation genetic testing for aneuploidy (NIPGT-A) is potentially appropriate to assess chromosomal ploidy of the embryo, practical application of it in a routine IVF center has not been started in the absence of a recommendation. We developed a comprehensive workflow for a clinically applicable strategy for NIPGT-A based on next-generation sequencing (NGS) technology. We performed MALBAC whole genome amplification and NGS on spent blastocyst culture media of Day 3 embryos fertilized with intra-cytoplasmic sperm injection (ICSI). Spent embryonic culture media of morphologically good quality score embryos were enrolled in further analysis with the blank culture media as background control. Chromosomal abnormalities were identified by an optimized bioinformatics pipeline applying a copy number variation (CNV) detecting algorithm. We demonstrate a comprehensive workflow covering both wet- and dry-lab procedures supporting a clinically applicable strategy for NIPGT-A. It can be carried out within 48 h which is critical for the same-cycle blastocyst transfer, but also suitable for “freeze all” and “elective frozen embryo” strategies. The described integrated approach of non-invasive evaluation of embryonic DNA content of the culture media can potentially supplement existing pre-implantation genetic screening methods.

Keywords: next generation sequencing, in vitro fertilization, embryo assessment, non-invasive pre-implantation genetic testing

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1072 Ex Vivo Permeation Comparison Study of Flurbiprofen from Nanoparticles through Human Skin

Authors: Sheimah El Bejjaji, Lara Gorsek, Chandler Quilchez, Joaquim Suñer, Mireia Mallandrich

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Flurbiprofen is an anti-inflammatory drug used in several treatments. The purpose of this study was to compare the permeation of two different formulations of flurbiprofen through the human skin. The first formulation was a solution of flurbiprofen dissolved with polyethylene glycol 3350 (PEG 3350). The second formulation was flurbiprofen encapsulated in poly-ɛ-caprolactone (PɛCL) nanoparticles (NPs), stabilized with poloxamer 188, submitted individually for freeze-drying with PEG 3350 as a cryoprotectant and sterilized by gamma-irradiation. Human skin was obtained from the abdominal region of a healthy patient. The experimental protocol was approved by the Bioethics Committee of Barcelona SCIAS Hospital (Spain), and they obtained the written informed consent forms. After being frozen to -20ºC, the skin samples were cut with a dermatome at 400 µm. The ex vivo permeation study was performed in Franz diffusion cells with a diffusion area of 2.54 cm². Skin samples were placed between two compartment sites, the dermal side in contact with the receptor medium and the epidermis side in contact with the donor chamber to which the formulation was applied. The permeation study was conducted for 24 hours at 32 ± 0.5 °C in accordance with sink conditions. The results were analyzed with an unpaired t-test, and the p-values indicate the formulation with nanoparticles had a higher permeability coefficient, flux, partition parameter, diffusion parameter, and lag time. The applicability of this formulation topically can benefit articulations and ligament inflammation as an alternative to oral drugs.

Keywords: anti-inflammatory drug, flurbiprofen, human skin, nanoparticles, skin permeation

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1071 Effect of Scrotal Circumference on Freezability of Bangladeshi Crossbred Bulls

Authors: Ajeet K. Jha, Pankaj K. Jha, Pravin Mishra

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The study was conducted to evaluate the freezability of crossbred bulls’ semen at early age. Semen of three consecutive collections at 7 days interval from 12 crossbred bulls 17 was evaluated. The age at first collection was 15 to 20 months. Evaluation of semen was done soon after collection. Triladyl, Minitub, Germany was used as extender and was frozen using standard semen freezing protocol. Post-thaw sperm motility was evaluated. Morphology of paraformaldehyde fixed spermatozoa was evaluated under differential interference phase contrast microscopy and the viability of spermatozoa was evaluated by using stain SYBR-14 (1 mM/ml) and propidium iodide (2.41 mM/ml) under an epifluorescent microscopy. Scrotal circumference was correlated with all possible measures in all groups of crossbred bulls. Volume of semen, sperm concentration, total number of spermatozoa, initial sperm motility, post-thaw sperm motility, proportion of normal spermatozoa and proportion of live spermatozoa were compared among individual bull within and between two groups of crossbred bulls. A significant positive correlation was observed between scrotal circumference and volume of semen and between scrotal circumference and the total number of sperm production per ejaculate (r = 0.72, p < 0.04). Significant variation was observed in different semen parameters among individual bulls within the same group (p < 0.05) but no significant variation was found between two groups of crossbred bulls. Out of 12 bulls, semen freezability of 10 bulls was found satisfactory while semen of 2 bulls (Local × Friesian) was unsatisfactory. In conclusion, crossbred bulls aged 18 months having scrotal circumference > 30 cm produce freezable quality semen.

Keywords: Bangladesh, crossbred bull, scrotal circumference, semen freezability

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1070 Effect of Degree of Phosphorylation on Electrospinning and In vitro Cell Behavior of Phosphorylated Polymers as Biomimetic Materials for Tissue Engineering Applications

Authors: Pallab Datta, Jyotirmoy Chatterjee, Santanu Dhara

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Over the past few years, phosphorous containing polymers have received widespread attention for applications such as high performance optical fibers, flame retardant materials, drug delivery and tissue engineering. Being pentavalent, phosphorous can exist in different chemical environments in these polymers which increase their versatility. In human biochemistry, phosphorous based compounds exert their functions both in soluble and insoluble form occurring as inorganic or as organophosphorous compounds. Specifically in case of biomacromolecules, phosphates are critical for functions of DNA, ATP, phosphoproteins, phospholipids, phosphoglycans and several coenzymes. Inspired by the role of phosphorous in functional biomacromolecules, design and synthesis of biomimetic materials are thus carried out by several authors to study macromolecular function or as substitutes in clinical tissue regeneration conditions. In addition, many regulatory signals of the body are controlled by phoshphorylation of key proteins present either in form of growth factors or matrix-bound scaffold proteins. This inspires works on synthesis of phospho-peptidomimetic amino acids for understanding key signaling pathways and this is extended to obtain molecules with potentially useful biological properties. Apart from above applications, phosphate groups bound to polymer backbones have also been demonstrated to improve function of osteoblast cells and augment performance of bone grafts. Despite the advantages of phosphate grafting, however, there is limited understanding on effect of degree of phosphorylation on macromolecular physicochemical and/or biological properties. Such investigations are necessary to effectively translate knowledge of macromolecular biochemistry into relevant clinical products since they directly influence processability of these polymers into suitable scaffold structures and control subsequent biological response. Amongst various techniques for fabrication of biomimetic scaffolds, nanofibrous scaffolds fabricated by electrospinning technique offer some special advantages in resembling the attributes of natural extracellular matrix. Understanding changes in physico-chemical properties of polymers as function of phosphorylation is therefore going to be crucial in development of nanofiber scaffolds based on phosphorylated polymers. The aim of the present work is to investigate the effect of phosphorous grafting on the electrospinning behavior of polymers with aim to obtain biomaterials for bone regeneration applications. For this purpose, phosphorylated derivatives of two polymers of widely different electrospinning behaviors were selected as starting materials. Poly(vinyl alcohol) is a conveniently electrospinnable polymer at different conditions and concentrations. On the other hand, electrospinning of chitosan backbone based polymers have been viewed as a critical challenge. The phosphorylated derivatives of these polymers were synthesized, characterized and electrospinning behavior of various solutions containing these derivatives was compared with electrospinning of pure poly (vinyl alcohol). In PVA, phosphorylation adversely impacted electrospinnability while in NMPC, higher phosphate content widened concentration range for nanofiber formation. Culture of MG-63 cells on electrospun nanofibers, revealed that degree of phosphate modification of a polymer significantly improves cell adhesion or osteoblast function of cultured cells. It is concluded that improvement of cell response parameters of nanofiber scaffolds can be attained as a function of controlled degree of phosphate grafting in polymeric biomaterials with implications for bone tissue engineering applications.

Keywords: bone regeneration, chitosan, electrospinning, phosphorylation

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1069 Needle Track Technique In Strabismus Surgery

Authors: Seema Dutt Bandhu, Yashi Bansal, Tania Moudgil, Barinder Kaur

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Introduction: Scleral perforation during the passage of suture needle is a known complication of strabismus surgery. The present study was conducted to evolve a safe and easy technique of passing the suture needle through the sclera. A scleral tunnel was created with a 26-guage needle through which the suture needle was passed. The rest of the steps of strabismus surgery were carried out as usual. Material and Methods: After taking clearance from the Institutional Ethics Committee, an interventional study was carried out on twenty patients. The scleral tunnel technique was performed on the patients of strabismus after taking written informed consent. Before passing the suture needle through the sclera during strabismus surgery, a tunnel through approximately half the thickness of the sclera was created with the help of a bent 26-gauge needle. The suture needle was then passed through this tunnel. Rest of the steps of the surgery were carried out in the conventional manner. In a control group of same number of patients, the surgery was performed in the conventional method. Both the groups were followed up for any complications. Ease of passing suture and surgeons’ satisfaction with the technique was noted on a 10-point Likert scale. Results: None of the patients in either group suffered from any complications. Four surgeons participated in the study. The average Likert scale score of the surgeons for satisfaction with the technique was 4.5 on a scale of 5. The score for ease of passage of suture needle was 5 on a score of 5. Discussion: Scleral perforation during passing the sutures through the sclera is a known complication of strabismus surgery. Incidence reported is 7.8% It occurs due to inappropriate engagement of the scleral tissue or passage of the suture needle along a wrong axis during the process of passing the suture needle. The needle track technique eases the passage of passing the suture needle through the sclera as the engagement of the scleral tissue can be done with greater control with a 26-guage needle. The surgeons have reported that they are highly satisfied with the technique and they have reported that the technique eased the passage of the suture needle through the sclera.

Keywords: suture, scleral tunnel, strabismus, scleral perforation

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1068 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

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Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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1067 Methylation Profiling and Validation of Candidate Tissue-Specific Differentially Methylated Regions for Identification of Human Blood, Saliva, Semen and Vaginal Fluid and Its Application in Forensics

Authors: Meenu Joshi, Natalie Naidoo, Farzeen Kader

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Identification of body fluids is an essential step in forensic investigation to aid in crime reconstruction. Tissue-specific differentially methylated regions (tDMRs) of the human genome can be targeted to be used as biomarkers to differentiate between body fluids. The present study was undertaken to establish the methylation status of potential tDMRs in blood, semen, saliva, and vaginal fluid by using methylation-specific PCR (MSP) and bisulfite sequencing (BS). The methylation statuses of 3 potential tDMRS in genes ZNF282, PTPRS, and HPCAL1 were analysed in 10 samples of each body fluid. With MSP analysis, the ZNF282, and PTPRS1 tDMR displayed semen-specific hypomethylation while HPCAL1 tDMR showed saliva-specific hypomethylation. With quantitative analysis by BS, the ZNF282 tDMR showed statistically significant difference in overall methylation between semen and all other body fluids as well as at individual CpG sites (p < 0.05). To evaluate the effect of environmental conditions on the stability of methylation profiles of the ZNF282 tDMR, five samples of each body fluid were subjected to five different forensic simulated conditions (dry at room temperature, wet in an exsiccator, outside on the ground, sprayed with alcohol, and sprayed with bleach) for 50 days. Vaginal fluid showed highest DNA recovery under all conditions while semen had least DNA quantity. Under outside on the ground condition, all body fluids except semen showed a decrease in methylation level; however, a significant decrease in methylation level was observed for saliva. A statistical significant difference was observed for saliva and semen (p < 0.05) for outside on the ground condition. No differences in methylation level were observed for the ZNF282 tDMR under all conditions for vaginal fluid samples. Thus, in the present study ZNF282 tDMR has been identified as a novel and stable semen-specific hypomethylation marker.

Keywords: body fluids, bisulphite sequencing, forensics, tDMRs, MSP

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1066 On the Thermodynamics of Biological Cell Adhesion

Authors: Ben Nadler

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Cell adhesion plays a vital role in many cell activities. The motivation to model cell adhesion is to study important biological processes, such as cell spreading, cell aggregation, tissue formation, and cell adhesion, which are very challenging to study by experimental methods alone. This study provides important insight into cell adhesion, which can lead to improve regenerative medicine and tissue formation techniques. In this presentation the biological cells adhesion is mediated by receptors–ligands binding and the diffusivity of the receptor on the cell membrane surface. The ability of receptors to diffuse on the cell membrane surface yields a very unique and complicated adhesion mechanism, which is exclusive to cells. The phospholipid bilayer, which is the main component in the cell membrane, shows fluid-like behavior associated with the molecules’ diffusivity. The biological cell is modeled as a fluid-like membrane with negligible bending stiffness enclosing the cytoplasm fluid. The in-plane mechanical behavior of the cell membrane is assumed to depend only on the area change, which is motivated by the fluidity of the phospholipid bilayer. In addition, the presence of receptors influences on the local mechanical properties of the cell membrane is accounted for by including stress-free area change, which depends on the receptor density. Based on the physical properties of the receptors and ligands the attraction between the receptors and ligands is modeled as a charged-nonpolar which is a noncovalent interaction. Such interaction is a short-range type, which decays fast with distance. The mobility of the receptor on the cell membrane is modeled using the diffusion equation and Fick’s law is used to model the receptor–receptor interactions. The resultant interaction force, which includes receptor–ligand and receptor–receptor interaction, is decomposed into tangential part, which governs the receptor diffusion, and normal part, which governs the cell deformation and adhesion. The formulation of the governing equations and numerical simulations will be presented. Analysis of the adhesion characteristic and properties are discussed. The roles of various thermomechanical properties of the cell, receptors and ligands on the cell adhesion are investigated.

Keywords: cell adhesion, cell membrane, receptor-ligand interaction, receptor diffusion

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1065 Increased Expression Levels of Soluble Epoxide Hydrolase in Obese and Its Modulation by Physical Exercise

Authors: Abdelkrim Khadir, Sina Kavalakatt, Preethi Cherian, Ali Tiss

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Soluble epoxide hydrolase (sEH) is an emerging therapeutic target in several chronic states that have inflammation as a common underlying cause such as immunometabolic diseases. Indeed, sEH is known to play a pro-inflammatory role by metabolizing anti-inflammatory, epoxyeicosatrienoic acids (EETs) to pro-inflammatory diols. Recently, it was shown sEH to be linked to diet and microbiota interaction in rat models of obesity. Nevertheless, the functional contribution of sEH and its anti-inflammatory substrates EETs in obesity remain poorly understood. In the current study, we compared the expression pattern of sEH between lean and obese nondiabetic human subjects using subcutaneous adipose tissue (SAT) and peripheral blood mononuclear cells (PBMCs). Using RT-PCR, western blot and immunofluorescence confocal microscopy, we show here that the level of sEH mRNA and protein to be significantly increased in obese subjects with concomitant increase in endoplasmic reticulum (ER) stress components (GRP78 and ATF6α) and inflammatory markers (TNF-α, IL-6) when compared to lean controls. The observation that sEH was overexpressed in obese subjects’ prompt us to investigate whether physical exercise could reduce its expression. In this study, we report here 3-months supervised physical exercise significantly attenuated the expression of sEH in both the SAT and PBMCs, with a parallel decrease in the expression of ER stress markers along with attenuated inflammatory response. On the other hand, homocysteine, a sulfur containing amino acid deriving from the essential amino acid methionine was shown to be directly associated with insulin resistance. When 3T3-L1 preadipocytes cells were treated with homocysteine our results show increased sEH levels along with ER stress markers. Collectively, our data suggest that sEH upregulation is strongly linked to ER stress in adiposity and that physical exercise modulates its expression. This gives further evidence that exercise might be useful as a strategy for managing obesity and preventing its associated complications.

Keywords: obesity, adipose tissue, epoxide hydrolase, ER stress

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1064 Prevalence of Lupus Glomerulonephritis in Renal Biopsies in an Eastern Region of the Arab World

Authors: M. Fayez Al Homsi, Reem Al Homsi

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Renal disease is a major cause of morbidity and mortality. Glomerular diseases make a small portion of the renal disease. Lupus glomerulonephritis (GN) is the commonest among the GN of systemic diseases. More than a hundred and eighty-eight consecutive renal biopsies are performed and evaluated for clinically suspected glomerular diseases over a period of two years. As in a standard practice after receiving the ultrasound-guided renal biopsies, the fresh biopsy is divided to three parts, one part is frozen for immunofluorescence evaluation, the second part is placed in 4% glutaraldehyde for electron microscopic evaluation, and the third part is placed in 10% buffered formalin for light microscopic evaluation. Primary glomerular diseases are detected in 83 biopsies; glomerulonephritis (GN) of systemic diseases are identified in 88, glomerular lesions in vascular diseases in 3, glomerular lesions in metabolic diseases in 7, hereditary nephropathies in 2, end-stage kidney in 2, and glomerular lesions in transplantation in 3 biopsies. Among the primary lesions, focal segmental glomerulosclerosis (28) and mesangial proliferative GN (26) were the most common. Lupus GN (67) and Ig A nephropathy (20) were the most common of the GN of systemic diseases. Lupus nephritis biopsies included one biopsy diagnosed as class 1 (normal), 17 biopsies class 2 (mesangial proliferation), 5 biopsies class 3 (focal proliferative GN), 39 biopsies class 4 diffuse proliferative GN), 3 biopsies class 5 (membranous GN), and 2 biopsies class 6 (crescentic GN). Lupus GN is the most common among GN of systemic diseases. While diabetes is very common here, diabetic GN (3 biopsies) is not as common as might one expects. Most likely this is due to sampling and reluctance on part of nephrologists and patients in sampling the kidney in diabetes mellitus.

Keywords: diabetes, glomerulonephritis, lupus, mesangial proliferation, nephropathy

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1063 Plasma Treatment in Conjunction with EGM-2 Medium Can Enhance Endothelial and Osteogenic Marker Expressions of Bone Marrow MSCs

Authors: Chih-Hsin Lin, Shyh-Yuan Lee, Yuan-Min Lin

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For many tissue engineering applications, an important goal is to create functional tissues in-vitro, and such tissues to be viable, they have to be vascularized. Endothelial cells (EC) and endothelial progenitor cells (EPC) are promising candidates for vascularization. However, both of them have limited expansion capacity and autologous cells currently do not exist for either ECs or EPCs. Therefore, we use bone marrow mesenchymal stem cells (MSC) as a source material for ECs. Growth supplements are commonly used to induce MSC differentiation, and further improvements in differentiation conditions can be made by modifying the cell's growth environment. An example is pre-treatment of the growth dish with gas plasma, in order to modify the surface functional groups of the material that the cells are seeded on. In this work, we compare the effects of different gas plasmas on the growth and differentiation of MSCs. We treat the dish with different plasmas (CO2, N2, and O2) and then induce MSC differentiation with endothelial growth medium-2 (EGM-2). We find that EGM-2 by itself upregulates EC marker CD31 mRNA expression, but not VEGFR2, CD34, or vWF. However, these additional EC marker expressions were increased for cells seeded on plasma treated substrates. Specifically, for EC markers, we found that N2 plasma treatment upregulated CD31 and VEGFR-2 mRNA expressions; CO2 plasma treatment upregulated CD34 and vWF mRNA expressions. The osteogenic markers ALP and osteopontin mRNA expressions were markedly enhanced on all plasma-treated dishes. We also found that plasma treatment in conjunction with EGM-2 growth medium can enhance MSCs differentiation into endothelial-like cells and osteogenic-like cells. Our work shows that the effect of the growth medium (EGM-2) on MSCs differentiation is influenced by the plasma modified surface chemistry of the substrate. In conclusion, plasma surface modification can enhance EGM-2 effectiveness and induced both endothelial and osteogenic differentiation. Our findings provide a method to enhance EGM-2 based cell differentiation, with consequences for tissue engineering and stem cell biology applications.

Keywords: endothelial differentiation, EGM-2, osteogenesis, plasma treatment, surface modification

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1062 Ill-Posed Inverse Problems in Molecular Imaging

Authors: Ranadhir Roy

Abstract:

Inverse problems arise in medical (molecular) imaging. These problems are characterized by large in three dimensions, and by the diffusion equation which models the physical phenomena within the media. The inverse problems are posed as a nonlinear optimization where the unknown parameters are found by minimizing the difference between the predicted data and the measured data. To obtain a unique and stable solution to an ill-posed inverse problem, a priori information must be used. Mathematical conditions to obtain stable solutions are established in Tikhonov’s regularization method, where the a priori information is introduced via a stabilizing functional, which may be designed to incorporate some relevant information of an inverse problem. Effective determination of the Tikhonov regularization parameter requires knowledge of the true solution, or in the case of optical imaging, the true image. Yet, in, clinically-based imaging, true image is not known. To alleviate these difficulties we have applied the penalty/modified barrier function (PMBF) method instead of Tikhonov regularization technique to make the inverse problems well-posed. Unlike the Tikhonov regularization method, the constrained optimization technique, which is based on simple bounds of the optical parameter properties of the tissue, can easily be implemented in the PMBF method. Imposing the constraints on the optical properties of the tissue explicitly restricts solution sets and can restore uniqueness. Like the Tikhonov regularization method, the PMBF method limits the size of the condition number of the Hessian matrix of the given objective function. The accuracy and the rapid convergence of the PMBF method require a good initial guess of the Lagrange multipliers. To obtain the initial guess of the multipliers, we use a least square unconstrained minimization problem. Three-dimensional images of fluorescence absorption coefficients and lifetimes were reconstructed from contact and noncontact experimentally measured data.

Keywords: constrained minimization, ill-conditioned inverse problems, Tikhonov regularization method, penalty modified barrier function method

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1061 Modification of Titanium Surfaces with Micro/Nanospheres for Local Antibiotic Release

Authors: Burcu Doymus, Fatma N. Kok, Sakip Onder

Abstract:

Titanium and titanium-based materials are commonly used to replace or regenerate the injured or lost tissues because of accidents or illnesses. Hospital infections and strong bond formation at the implant-tissue interface are directly affecting the success of the implantation as weak bonding with the native tissue and hospital infections lead to revision surgery. The purpose of the presented study is to modify the surface of the titanium substrates with nano/microspheres for local drug delivery and to prevent hospital infections. Firstly, titanium surfaces were silanized with APTES (3-Triethoxysilylpropylamine) following the negatively charged oxide layer formation. Then characterization studies using Scanning Electron Microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) were done on the modified surfaces. Secondly, microspheres/nanospheres were prepared with chitosan that is a natural polymer and having valuable properties such as non-toxicity, high biocompatibility, low allergen city and biodegradability for biomedical applications. Antibiotic (ciprofloxacin) loaded micro/nanospheres have been fabricated using emulsion cross-linking method and have been immobilized onto the titanium surfaces with different immobilization techniques such as covalent bond and entrapment. Optimization studies on size and drug loading capacities of micro/nanospheres were conducted before the immobilization process. Light microscopy and SEM were used to visualize and measure the size of the produced micro/nanospheres. Loaded and released drug amounts were determined by using UV- spectrophotometer at 278 nm. Finally, SEM analysis and drug release studies on the micro/nanospheres coated Ti surfaces were done. As a conclusion, it was shown that micro/nanospheres were immobilized onto the surfaces successfully and drug release from these surfaces was in a controlled manner. Moreover, the density of the micro/nanospheres after the drug release studies was higher on the surfaces where the entrapment technique was used for immobilization. Acknowledgement: This work is financially supported by The Scientific and Technological Research Council Of Turkey (Project # 217M220)

Keywords: chitosan, controlled drug release, nanosphere, nosocomial infections, titanium

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1060 Biomaterials Solutions to Medical Problems: A Technical Review

Authors: Ashish Thakur

Abstract:

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

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1059 Alternate Optical Coherence Tomography Technologies in Use for Corneal Diseases Diagnosis in Dogs and Cats

Authors: U. E. Mochalova, A. V. Demeneva, Shilkin A. G., J. Yu. Artiushina

Abstract:

Objective. In medical ophthalmology OCT has been actively used in the last decade. It is a modern non-invasive method of high-precision hardware examination, which gives a detailed cross-sectional image of eye tissues structure with a high level of resolution, which provides in vivo morphological information at the microscopic level about corneal tissue, structures of the anterior segment, retina and optic nerve. The purpose of this study was to explore the possibility of using the OCT technology in complex ophthalmological examination in dogs and cats, to characterize the revealed pathological structural changes in corneal tissue in cats and dogs with some of the most common corneal diseases. Procedures. Optical coherence tomography of the cornea was performed in 112 animals: 68 dogs and 44 cats. In total, 224 eyes were examined. Pathologies of the organ of vision included: dystrophy and degeneration of the cornea, endothelial corneal dystrophy, dry eye syndrome, chronic superficial vascular keratitis, pigmented keratitis, corneal erosion, ulcerative stromal keratitis, corneal sequestration, chronic glaucoma and also postoperative period after performed keratoplasty. When performing OCT, we used certified medical devices: "Huvitz HOCT-1/1F», «Optovue iVue 80» and "SOCT Copernicus Revo (60)". Results. The results of a clinical study on the use of optical coherence tomography (OCT)of the cornea in cats and dogs, performed by the authors of the article in the complex diagnosis of keratopathies of variousorigins: endothelial corneal dystrophy, pigmented keratitis, chronic keratoconjunctivitis, chronic herpetic keratitis, ulcerative keratitis, traumatic corneal damage, sequestration of the cornea of cats, chronic keratitis, complicating the course of glaucoma. The characteristics of the OCT scans are givencorneas of cats and dogs that do not have corneal pathologies. OCT scans of various corneal pathologies in dogs and cats with a description of the revealed pathological changes are presented. Of great clinical interest are the data obtained during OCT of the cornea of animals undergoing keratoplasty operations using various forms of grafts. Conclusions. OCT makes it possible to assess the thickness and pathological structural changes of the corneal surface epithelium, corneal stroma and descemet membrane. We can measure them, determine the exact localization, and record pathological changes. Clinical observation of the dynamics of the pathological process in the cornea using OCT makes it possible to evaluate the effectiveness of drug treatment. In case of negative dynamics of corneal disease, it is necessary to determine the indications for surgical treatment (to assess the thickness of the cornea, the localization of its thinning zones, to characterize the depth and area of pathological changes). According to the OCT of the cornea, it is possible to choose the optimal surgical treatment for the patient, the technique and depth of optically constructive surgery (penetrating or anterior lamellar keratoplasty).; determine the depth and diameter of the planned microsurgical trepanation of corneal tissue, which will ensure good adaptation of the edges of the donor material.

Keywords: optical coherence tomography, corneal sequestration, optical coherence tomography of the cornea, corneal transplantation, cat, dog

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1058 Protective Effect of N-Acetyl Cysteine and Alpha Lipoic Acid on Rats Chronically Exposed to Cadmium Chloride

Authors: S. El Ballal, H. El Sabbagh, M. Abd El Gaber, A. Eisa, A. Al Gamal

Abstract:

Cadmium is one of the most harmful heavy metals able to induce severe injury. In this study, sixty four male Sprague Dawley rats weighing (70-80 gm) were used. Rats were divided into 4 groups each group of 16 rats. Group A: served as control and received commercial ration and distilled water Group B: cadmium chloride was administered orally in water at dose of 300 ppm cadmium (560 mg/L as CdCl2). Group C: Animals received cadmium in drinking water in addition to administration of N-acetylcysteine (NAC) orally at a dose of 150 mg/kg body weight, equivalent to 1500 ppm in food. Group D: Animals received cadmium in drinking water in addition to administration of alpha lipoic acid (ALA) orally at a dose of 150 mg/kg body weight, equivalent to 1500 ppm in food. The experiment was continued for 2 months. Collection of blood and tissue samples was performed at 2, 4, 6, 8 weeks. Blood sample were collected for serum biochemical analysis including malondialdehyde (MDA), total antioxidants, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, urea and uric acid. Tissue specimens were collected for histopathological examination including liver, kidney, brain and testis. Histopathological examination revealed that cadmium choloride induces pathological alterations which increased in severity with time. The use of NAC and ALA can ameliorate toxic effect of CdCl2. The results showed significant decrease MDA and significant increase total antioxidants in group C and D compared to group B, Liver enzymes include AST and ALT showed significant decrease. Regarding to results of total protein and albumin, they revealed significant increase. Urea and uric acid showed significant decrease. From our study we conclude that NAC and ALA have protective effect against cadmium toxicity.

Keywords: ALA, cadmium, histopathology, NAC

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1057 Inflammatory and Cardio Hypertrophic Remodeling Biomarkers in Patients with Fabry Disease

Authors: Margarita Ivanova, Julia Dao, Andrew Friedman, Neil Kasaci, Rekha Gopal, Ozlem Goker-Alpan

Abstract:

In Fabry disease (FD), α-galactosidase A (α-Gal A) deficiency leads to the accumulation of globotriaosylceramide (Lyso-Gb3 and Gb3), triggering a pathologic cascade that causes the severity of organs damage. The heart is one of the several organs with high sensitivity to the α-Gal A deficiency. A subgroup of patients with significant residual of α-Gal A activity with primary cardiac involvement is occasionally referred to as “cardiac variant.” The cardiovascular complications are most frequently encountered, contributing substantially to morbidity, and are the leading cause of premature death in male and female patients with FD. The deposition of Lyso-Gb-3 and Gb-3 within the myocardium affects cardiac function with resultant progressive cardiovascular pathology. Gb-3 and Lyso-Gb-3 accumulation at the cellular level trigger a cascade of events leading to end-stage fibrosis. In the cardiac tissue, Lyso-Gb-3 deposition is associated with the increased release of inflammatory factors and transforming growth factors. Infiltration of lymphocytes and macrophages into endomyocardial tissue indicates that inflammation plays a significant role in cardiac damage. Moreover, accumulated data suggest that chronic inflammation leads to multisystemic FD pathology even under enzyme replacement therapy (ERT). NF-κB activation plays a subsequent role in the inflammatory response to cardiac dysfunction and advanced heart failure in the general population. TNFalpha/NF-κB signaling protects the myocardial evoking by ischemic preconditioning; however, this protective effect depends on the concentration of TNF-α. Thus, we hypothesize that TNF-α is a critical factor in determining the grade of cardio-pathology. Cardiac hypertrophy corresponds to the expansion of the coronary vasculature to maintain a sufficient supply of nutrients and oxygen. Coronary activation of angiogenesis and fibrosis plays a vital role in cardiac vascularization, hypertrophy, and tissue remodeling. We suggest that the interaction between the inflammatory pathways and cardiac vascularization is a bi-directional process controlled by secreted cytokines and growth factors. The co-coordination of these two processes has never been explored in FD. In a cohort of 40 patients with FD, biomarkers associated with inflammation and cardio hypertrophic remodeling were studied. FD patients were categorized into three groups based on LVmass/DSA, LVEF, and ECG abnormalities: FD with no cardio complication, FD with moderate cardio complication, and severe cardio complication. Serum levels of NF-kB, TNFalpha, Il-6, Il-2, MCP1, ING-gamma, VEGF, IGF-1, TGFβ, and FGF2 were quantified by enzyme-linked immunosorbent assays (ELISA). Among the biomarkers, MCP-1, INF-gamma, VEGF, TNF-alpha, and TGF-beta were elevated in FD patients. Some of these biomarkers also have the potential to correlate with cardio pathology in FD. Conclusion: The study provides information about the role of inflammatory pathways and biomarkers of cardio hypertrophic remodeling in FD patients. This study will also reveal the mechanisms that link intracellular accumulation of Lyso-GB-3 and Gb3 to the development of cardiomyopathy with myocardial thickening and resultant fibrosis.

Keywords: biomarkers, Fabry disease, inflammation, growth factors

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1056 Calibration of Mini TEPC and Measurement of Lineal Energy in a Mixed Radiation Field Produced by Neutrons

Authors: I. C. Cho, W. H. Wen, H. Y. Tsai, T. C. Chao, C. J. Tung

Abstract:

Tissue-equivalent proportional counter (TEPC) is a useful instrument used to measure radiation single-event energy depositions in a subcellular target volume. The quantity of measurements is the microdosimetric lineal energy, which determines the relative biological effectiveness, RBE, for radiation therapy or the radiation-weighting factor, WR, for radiation protection. TEPC is generally used in a mixed radiation field, where each component radiation has its own RBE or WR value. To reduce the pile-up effect during radiotherapy measurements, a miniature TEPC (mini TEPC) with cavity size in the order of 1 mm may be required. In the present work, a homemade mini TEPC with a cylindrical cavity of 1 mm in both the diameter and the height was constructed to measure the lineal energy spectrum of a mixed radiation field with high- and low-LET radiations. Instead of using external radiation beams to penetrate the detector wall, mixed radiation fields were produced by the interactions of neutrons with TEPC walls that contained small plugs of different materials, i.e. Li, B, A150, Cd and N. In all measurements, mini TEPC was placed at the beam port of the Tsing Hua Open-pool Reactor (THOR). Measurements were performed using the propane-based tissue-equivalent gas mixture, i.e. 55% C3H8, 39.6% CO2 and 5.4% N2 by partial pressures. The gas pressure of 422 torr was applied for the simulation of a 1 m diameter biological site. The calibration of mini TEPC was performed using two marking points in the lineal energy spectrum, i.e. proton edge and electron edge. Measured spectra revealed high lineal energy (> 100 keV/m) peaks due to neutron-capture products, medium lineal energy (10 – 100 keV/m) peaks from hydrogen-recoil protons, and low lineal energy (< 10 keV/m) peaks of reactor photons. For cases of Li and B plugs, the high lineal energy peaks were quite prominent. The medium lineal energy peaks were in the decreasing order of Li, Cd, N, A150, and B. The low lineal energy peaks were smaller compared to other peaks. This study demonstrated that internally produced mixed radiations from the interactions of neutrons with different plugs in the TEPC wall provided a useful approach for TEPC measurements of lineal energies.

Keywords: TEPC, lineal energy, microdosimetry, radiation quality

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1055 The Importance of including All Data in a Linear Model for the Analysis of RNAseq Data

Authors: Roxane A. Legaie, Kjiana E. Schwab, Caroline E. Gargett

Abstract:

Studies looking at the changes in gene expression from RNAseq data often make use of linear models. It is also common practice to focus on a subset of data for a comparison of interest, leaving aside the samples not involved in this particular comparison. This work shows the importance of including all observations in the modeling process to better estimate variance parameters, even when the samples included are not directly used in the comparison under test. The human endometrium is a dynamic tissue, which undergoes cycles of growth and regression with each menstrual cycle. The mesenchymal stem cells (MSCs) present in the endometrium are likely responsible for this remarkable regenerative capacity. However recent studies suggest that MSCs also plays a role in the pathogenesis of endometriosis, one of the most common medical conditions affecting the lower abdomen in women in which the endometrial tissue grows outside the womb. In this study we compared gene expression profiles between MSCs and non-stem cell counterparts (‘non-MSC’) obtained from women with (‘E’) or without (‘noE’) endometriosis from RNAseq. Raw read counts were used for differential expression analysis using a linear model with the limma-voom R package, including either all samples in the study or only the samples belonging to the subset of interest (e.g. for the comparison ‘E vs noE in MSC cells’, including only MSC samples from E and noE patients but not the non-MSC ones). Using the full dataset we identified about 100 differentially expressed (DE) genes between E and noE samples in MSC samples (adj.p-val < 0.05 and |logFC|>1) while only 9 DE genes were identified when using only the subset of data (MSC samples only). Important genes known to be involved in endometriosis such as KLF9 and RND3 were missed in the latter case. When looking at the MSC vs non-MSC cells comparison, the linear model including all samples identified 260 genes for noE samples (including the stem cell marker SUSD2) while the subset analysis did not identify any DE genes. When looking at E samples, 12 genes were identified with the first approach and only 1 with the subset approach. Although the stem cell marker RGS5 was found in both cases, the subset test missed important genes involved in stem cell differentiation such as NOTCH3 and other potentially related genes to be used for further investigation and pathway analysis.

Keywords: differential expression, endometriosis, linear model, RNAseq

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1054 Transcranial Electric Field Treatments on Redox-Toxic Iron Deposits in Transgenic Alzheimer’s Disease Mouse Models: The Electroceutical Targeting of Alzheimer’s Disease

Authors: Choi Younshick, Lee Wonseok, Lee Jaemeun, Park Sun-Hyun, Kim Sunwoung, Park Sua, Kim Eun Ho, Kim Jong-Ki

Abstract:

Iron accumulation in the brain accelerates Alzheimer’s disease progression. To cure iron toxicity, we assessed the therapeutic effects of noncontact transcranial electric field stimulation to the brain on toxic iron deposits in either the Aβ-fibril structure or the Aβ plaque in a mouse model of Alzheimer’s disease (AD). A capacitive electrode-based alternating electric field (AEF) was applied to a suspension of magnetite (Fe₃O₄) to measure the field-sensitized electro-Fenton effect and resultant reactive oxygen species (ROS) generation. The increase in ROS generation compared to the untreated control was both exposure-time and AEF-frequency dependent. The frequency-specific exposure of AEF to 0.7–1.4 V/cm on a magnetite-bound Aβ-fibril or a transgenic Alzheimer’s disease (AD) mouse model revealed the removal of intraplaque ferrous magnetite iron deposit and Aβ-plaque burden together at the same time compared to the untreated control. The results of the behavioral tests show an improvement in impaired cognitive function following AEF treatment on the AD mouse model. Western blot assay found some disease-modifying biological responses, including down-regulating ferroptosis, neuroinflammation and reactive astrocytes that eventually made cognitive improvement feasible. Tissue clearing and 3D-imaging analysis revealed no induced damage to the neuronal structures of normal brain tissue following AEF treatment. In conclusion, our results suggest that the effective degradation of magnetite-bound amyloid fibrils or plaques in the AD brain by the electro-Fenton effect from electric field-sensitized magnetite offers a potential electroceutical treatment option for AD.

Keywords: electroceutical, intraplaque magnetite, alzheimer’s disease, transcranial electric field, electro-fenton effect

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1053 Different Feedings on Chemical Characteristics of Atlantic Salmon Fillet

Authors: Mahsa Jalili, Trude Johansen, Signe Dille Lovmo, Turid Rustad, Rolf Erik Olsen, Atle M. Bones

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The quality of fish muscle is a key factor in fish industry, and dietary ingredients can influence fish quality. The aim of this study was to examine the impact of krill meal, soybean meal, Bactocell® and butyrate fortified feeds and control diet on characteristics of salmon fillet. Thirty Atlantic salmon (6 per each group) were farmed for 12 weeks. All the fish were killed and frozen immediately. The white muscle from top posterior part of dorsal fin was dissected to analyze fat content, carotenoid content, content of water-soluble and salt-soluble proteins, cathepsin B and cathepsin B-L activities. ANOVA test was used to analyze mean and standard error of mean values at 0.05 significance level. There were significant difference in cathepsin B activity, water-soluble proteins and salt-soluble proteins (p-value= 0.005, 0.009 and 0.002). The mean values of other factors were not significantly different among the groups. Cathepsin B activity was higher in soymeal group. Water-soluble proteins were reported higher in soy meal and krill groups and salt-soluble proteins were significantly higher in soy meal and butyrate rich diets. Although soy meal has proven effect on enteritis, it results in higher percentage of protein in fillets. On the other hand, this feeding may have role in textural deterioration of fillets owing to higher values of endogenous cathepsin B in soymeal group.

Keywords: aquaculture, food quality, Krill protein extract, prebiotics, probiotics, Salmo salar, soy

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1052 The Modulation of Health and Inflammatory Status in Young Pigs by Grape Waste Enriched in Polyphenols

Authors: Gina Cecilia Pistol, Loredana Calin, Mariana Stancu, Veronica Chedea, Ionelia Taranu

Abstract:

Inflammatory-associated diseases have an increased trend in the past decades. The pharmacological strategies aimed to treat these inflammatory diseases are very expensive and with non-beneficial results. The current trend is to find alternative strategies to counteract or to control inflammatory component of diseases. The grape by-products either seeds or pomace are rich in bioactive compounds (e.g. polyphenols) which may be beneficial in prevention of inflammation associated with cancer progression and other pathologies with inflammatory component. The in vivo models are very useful for studying the immune and inflammatory status. The domestic pig (Sus scrofa domesticus) is related to human from anatomic and physiologic point of view, representing a feasible model for studying the human inflammatory pathologies. Starting from these data, we evaluated the effect of a diet containing 5% grape seed cakes (GS) on piglets blood biochemical parameters and immune pro- and anti-inflammatory biomarkers (IL-1 beta, IL-8, TNF-alpha, IL-6, IFN-gamma, IL-10, IL-4) in spleen and lymph nodes. 12 weaned piglets were fed for 30 days with a control diet or an experimental diet containing 5% GS. At the end of trial, plasma and tissue samples (spleen and lymph nodes) were collected and the biochemical and inflammatory markers were analysed by using biochemistry analyser and ELISA techniques. Our results showed that diet included 5% GS did not influence the health status determined by plasma biochemical parameters. Only a tendency for a slight increase of the biochemical parameters associated with energetic profile (glucose, cholesterol, triglycerides) was observed. Also, GS diet had no effect on pro- and anti-inflammatory cytokines content in spleen and lymph nodes tissue. Further experiments are needed in order to investigate other rate of dietary inclusion which could provide more evidence about the effect of grape bioactive compounds on pigs used as animal model.

Keywords: animal model, inflammation, grape seed by-product, immune organs

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1051 Influence of the Cooking Technique on the Iodine Content of Frozen Hake

Authors: F. Deng, R. Sanchez, A. Beltran, S. Maestre

Abstract:

The high nutritional value associated with seafood is related to the presence of essential trace elements. Moreover, seafood is considered an important source of energy, proteins, and long-chain polyunsaturated fatty acids. Generally, seafood is consumed cooked. Consequently, the nutritional value could be degraded. Seafood, such as fish, shellfish, and seaweed, could be considered as one of the main iodine sources. The deficient or excessive consumption of iodine could cause dysfunction and pathologies related to the thyroid gland. The main objective of this work is to evaluated iodine stability in hake (Merluccius) undergone different culinary techniques. The culinary process considered were: boiling, steaming, microwave cooking, baking, cooking en papillote (twisted cover with the shape of a sweet wrapper) and coating with a batter of flour and deep-frying. The determination of iodine was carried by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Regarding sample handling strategies, liquid-liquid extraction has demonstrated to be a powerful pre-concentration and clean-up approach for trace metal analysis by ICP techniques. Extraction with tetramethylammonium hydroxide (TMAH reagent) was used as a sample preparation method in this work. Based on the results, it can be concluded that the stability of iodine was degraded with the cooking processes. The major degradation was observed for the boiling and microwave cooking processes. The content of iodine in hake decreased up to 60% and 52%, respectively. However, if the boiling cooking liquid is preserved, this loss that has been generated during cooking is reduced. Only when the fish was cooked by following the cooking en papillote process the iodine content was preserved.

Keywords: cooking process, ICP-MS, iodine, hake

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1050 Open Fields' Dosimetric Verification for a Commercially-Used 3D Treatment Planning System

Authors: Nashaat A. Deiab, Aida Radwan, Mohamed Elnagdy, Mohamed S. Yahiya, Rasha Moustafa

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This study is to evaluate and investigate the dosimetric performance of our institution's 3D treatment planning system, Elekta PrecisePLAN, for open 6MV fields including square, rectangular, variation in SSD, centrally blocked, missing tissue, square MLC and MLC shaped fields guided by the recommended QA tests prescribed in AAPM TG53, NCS report 15 test packages, IAEA TRS 430 and ESTRO booklet no.7. The study was performed for Elekta Precise linear accelerator designed for clinical range of 4, 6 and 15 MV photon beams with asymmetric jaws and fully integrated multileaf collimator that enables high conformance to target with sharp field edges. Seven different tests were done applied on solid water equivalent phantom along with 2D array dose detection system, the calculated doses using 3D treatment planning system PrecisePLAN, compared with measured doses to make sure that the dose calculations are accurate for open fields including square, rectangular, variation in SSD, centrally blocked, missing tissue, square MLC and MLC shaped fields. The QA results showed dosimetric accuracy of the TPS for open fields within the specified tolerance limits. However large square (25cm x 25cm) and rectangular fields (20cm x 5cm) some points were out of tolerance in penumbra region (11.38 % and 10.9 %, respectively). For the test of SSD variation, the large field resulted from SSD 125 cm for 10cm x 10cm filed the results recorded an error of 0.2% at the central axis and 1.01% in penumbra. The results yielded differences within the accepted tolerance level as recommended. Large fields showed variations in penumbra. These differences between dose values predicted by the TPS and the measured values at the same point may result from limitations of the dose calculation, uncertainties in the measurement procedure, or fluctuations in the output of the accelerator.

Keywords: quality assurance, dose calculation, 3D treatment planning system, photon beam

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