Search results for: nerve differentiation

Authors: Alireza Shams, Ali Zamanian, Atefehe Shamosi, Farnaz Ghorbani

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Introduction: Although the application of a new strategy remains a remarkable challenge for treatment of disabilities due to neuronal defects, progress in Nanomedicine and tissue engineering, suggesting the new medical methods. One of the promising strategies for reconstruction and regeneration of nervous tissue is replacing of lost or damaged cells by specific scaffolds after Compressive, ischemic and traumatic injuries of central nervous system. Furthermore, ultrastructure, composition, and arrangement of tissue scaffolds are effective on cell grafts. We followed implantation and differentiation of mesenchyme stem cells to neural cells on Gelatin Polylactic-co-glycolic acid (PLGA) scaffolds coated with iron nanoparticles. The aim of this study was to evaluate the capability of stem cells to differentiate into motor neuron-like cells under topographical cues and morphogenic factors. Methods and Materials: Bone marrow mesenchymal stem cells (BMMSCs) was obtained by primary cell culturing of adult rat bone marrow got from femur bone by flushing method. BMMSCs were incubated with DMEM/F12 (Gibco), 15% FBS and 100 U/ml pen/strep as media. Then, BMMSCs seeded on Gel/PLGA scaffolds and tissue culture (TCP) polystyrene embedded and incorporated by Fe Nano particles (FeNPs) (Fe3o4 oxide (M w= 270.30 gr/mol.). For neuronal differentiation, 2×10 5 BMMSCs were seeded on Gel/PLGA/FeNPs scaffolds was cultured for 7 days and 0.5 µ mol. Retinoic acid, 100 µ mol. Ascorbic acid,10 ng/ml. Basic fibroblast growth factor (Sigma, USA), 250 μM Iso butyl methyl xanthine, 100 μM 2-mercaptoethanol, and 0.2 % B27 (Invitrogen, USA) added to media. Proliferation of BMMSCs was assessed by using MTT assay for cell survival. The morphology of BMMSCs and scaffolds was investigated by scanning electron microscopy analysis. Expression of neuron-specific markers was studied by immunohistochemistry method. Data were analyzed by analysis of variance, and statistical significance was determined by Turkey’s test. Results: Our results revealed that differentiation and survival of BMMSCs into motor neuron-like cells on Gel/PLGA/FeNPs as a biocompatible and biodegradable scaffolds were better than those cultured in Gel/PLGA in absence of FeNPs and TCP scaffolds. FeNPs had raised physical power but decreased capacity absorption of scaffolds. Well defined oriented pores in scaffolds due to FeNPs may activate differentiation and synchronized cells as a mechanoreceptor. Induction effects of magnetic FeNPs by One way flow of channels in scaffolds help to lead the cells and can facilitate direction of their growth processes. Discussion: Progression of biological properties of BMMSCs and the effects of FeNPs spreading under magnetic field was evaluated in this investigation. In vitro study showed that the Gel/PLGA/FeNPs scaffold provided a suitable structure for motor neuron-like cells differentiation. This could be a promising candidate for enhancing repair and regeneration in neural defects. Dynamic and static magnetic field for inducing and construction of cells can provide better results for further experimental studies.

Keywords: differentiation, mesenchymal stem cells, nano particles, neuronal defects, Scaffolds

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Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

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Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

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Authors: Joseph Roberson, Carina Kleynhans, Willie Coetzee

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The low entry barriers of the restaurant industry lead to an extremely competitive business environment. In this volatile business sector it is of the utmost importance to implement a strategy of quality differentiation. Vital aspects of a quality differentiation strategy are total quality management, benchmarking and service quality management. Ultimately, restaurant success depends on the continuous support of customers. Customers select restaurants based on their expectations of quality. If the customers' expectations are met, they perceive quality service and will re-patronize the restaurant. The restaurateur can manage perceptions of quality by influencing expectations while ensuring that those expectations are not inflated. The management of expectations can be done by communicating service quality to customers. The aim of this research paper is to describe the development of a grading process for restaurants. An assessment of the extensive body of literature on grading was conducted through content analysis. A standardized method for developing a grading system would assist in successful grading systems that could inform both customers and restaurateurs of restaurant quality.

Keywords: benchmarking, restaurants, grading, service quality, total quality management

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Authors: Manoj Deepak, N. Mathivanan, K. Venkatachalam

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Nerve root stenosis is one of the main cause for back pain. There are many methods both conservative and surgical to treat this disease. It is pertinent to decompress the spine to a proper extent so as to avoid the recurrence of symptoms. But too much of an aggressive approach also has its disadvantages. We present one of the methods to effectively decompress the nerve with better results. Our study was carried out in 52 patients with foramina stenosis between 2008 to 2011.We carried out the surgical procedure of shaving off the medial part of the facet joint so as to decompress the root. We selected those patients who had symptoms of claudication for more than 2 years. They had no signs of instability and they underwent conservative treatment for a period of 2 months before the procedure. Oswersty scoring was used to record the functional level of the patient before and after the procedure. All patients were followed up for a period of minimum 2.5 years. After evaluation for a minimum of 2.5 years, 34 patients had no evidence of recurrence of symptoms with improvement in the functional level.7 patients complained of minimal pain but their functional quality had improved postop. Six patients had symptoms of lumbar canal disease which reduced with conservative treatment. 5 patients required spinal fusion surgeries in the later period. Conclusion: Thus, we can effectively conclude that our procedure is safe and effective in reducing the symptoms in those patients with neurogenic claudication.

Keywords: facetectoemy, stenosis, decompression, Lumbar Foraminal Stenosis, hemi-facetectomy

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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: Jasna Hrncic

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Poor family relations predict depression, but also to other mental health issues. Mediating effect of self-efficacy and differentiation of self and moderating effect of decreased accessibility and/or success of other adaptive and defensive mechanisms for overcoming social disadvantages could explain depression as a specific outcome of dysfunctional family relations. The present study analyzes the mediation effect of self-efficacy and differentiation of self from poor family functioning to depressive symptoms and the moderation effect of deprivation of liberty on the listed mediation effect. Deprivation of liberty has, as a general consequence, a decreased accessibility and/or success of many adaptive and defensive mechanisms. It is hypothesized that: 1) self-efficacy and differentiation of self will mediate between family functioning and depressiveness in the total sample, and 2) deprivation of liberty will moderate the stated relations. Cross-sectional study was conducted among 323 male juveniles in Serbia divided in three groups: 98 adolescents deprived of their liberty due to antisocial behavior (incarcerated antisocial group - IAG), 121 adolescents with antisocial behavior in their natural setting (antisocial control group - CAG) and 105 adolescents in general population (general control group - CGG). The CAG was included along with GCG to control the possible influence that comorbidity of antisocial behavior and depressiveness could have on results. Instruments for family relations assessment were: for a whole family of origin the emotional exchange scale and individuation scale from GRADIR by Knezevic, and for a relationship with mother PCS-YSR and CRPBI by barber, and intimacy, rejection, sacrifice, punishment, demands, control and internal control by Opacic and Kos. Differentiation of self (DOS) is measured by emotional self scale (Opacic), self-efficacy (SE) by general incompetence scale by Bezinovic, and depression by BDI (Back), CES-D (Radloff) and D6R (Momirovic). Two-path structural equation modeling based on most commonly reported fit indices, showed that the mediation model had unfavorable fit to our data for total sample [(χ2 (1, N = 324) = 13.73); RMSEA= .20 (90% CI= [.12, .30]); CFI= .98; NFI= .97; AIC=31.73]. Path model provided an adequate fit to the data only for AIG - and not to the data from ACG and GCG. SE and DOS mediated the relationship between PFF and depressiveness. Test of the indirect effects revealed that 23.85% of PFF influences on depressiveness is mediated by these two mediators (the quotient of mediated effect = .24). Test of specific indirect effects showed that SE mediates 22.17%, while DOS mediates 1.67% of PFF influence on depressiveness. Lack of expected mediation effect could be explained by missing other potential mediators (i.e., relationship with that father, social skills, self-esteem) and lower variability of both predictor and criterion variable due to their low levels on the whole sample and on control subsamples. Results suggested that inaccessibility and/or successfulness of other adaptive and defensive mechanisms for overcoming social disadvantages has a strong impact on the mediation effect of self/efficacy and differentiation of self from poor family functioning to depressive symptoms. Further researches could include other potential mediators and a sample of clinically depressed people.

Keywords: antisocial behavior, mediating effect, moderating effect, natural setting, incarceration

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Authors: Deborah Zelinsky

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The retina filters millions of incoming signals into a smaller amount of exiting optic nerve fibers that travel to different portions of the brain. Most of the signals are for eyesight (called "image-forming" signals). However, there are other faster signals that travel "elsewhere" and are not directly involved with eyesight (called "non-image-forming" signals). This article centers on the neurons of the optic nerve connecting to parts of the limbic system. Eye care providers are currently looking at parvocellular and magnocellular processing pathways without realizing that those are part of an enormous "galaxy" of all the body systems. Lenses are modifying both non-image and image-forming pathways, taking A.M. Skeffington's seminal work one step further. Almost 100 years ago, he described the Where am I (orientation), Where is It (localization), and What is It (identification) pathways. Now, among others, there is a How am I (animation) and a Who am I (inclination, motivation, imagination) pathway. Classic eye testing considers pupils and often assesses posture and motion awareness, but classical prescriptions often overlook limbic involvement in visual processing. The limbic system is composed of the hippocampus, amygdala, hypothalamus, and anterior nuclei of the thalamus. The optic nerve's limbic connections arise from the intrinsically photosensitive retinal ganglion cells (ipRGC) through the "retinohypothalamic tract" (RHT). There are two main hypothalamic nuclei with direct photic inputs. These are the suprachiasmatic nucleus and the paraventricular nucleus. Other hypothalamic nuclei connected with retinal function, including mood regulation, appetite, and glucose regulation, are the supraoptic nucleus and the arcuate nucleus. The retino-hypothalamic tract is often overlooked when we prescribe eyeglasses. Each person is different, but the lenses we choose are influencing this fast processing, which affects each patient's aiming and focusing abilities. These signals arise from the ipRGC cells that were only discovered 20+ years ago and do not address the campana retinal interneurons that were only discovered 2 years ago. As eyecare providers, we are unknowingly altering such factors as lymph flow, glucose metabolism, appetite, and sleep cycles in our patients. It is important to know what we are prescribing as the visual processing evaluations expand past the 20/20 central eyesight.

Keywords: neuromodulation, retinal processing, retinohypothalamic tract, limbic system, visual processing

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Authors: Sundip Charmode, Simmi Mehra, Sudhir Kushwaha, Shalom Philip, Pratik Amrutiya, Ranjna Jangal

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Introduction: Quadrangular space syndrome involves compression of the axillary nerve and posterior circumflex humeral artery and its management in few cases, requires surgical decompression. The current study reviews the surgical approaches used in the decompression of neurovascular structures and presents our reflections and recommendations. Methods: Four human cadavers, in the Department of Anatomy were used for dissection of the Axillae and the Scapular region by the senior residents of the Department of Anatomy and Department of Orthopedics, who dissected quadrangular space in the eight upper limbs, using anterior and posterior surgical approaches. Observations: Posterior approach to identify the quadrangular space and secure its contents was recognized as the easier and much quicker method by both the Anatomy and Orthopedic residents, but it may result in increased postoperative morbidity. Whereas the anterior (Delto-pectoral) approach involves more skill but reduces postoperative morbidity. Conclusions: Anterior (Delto-pectoral) approach with suggested modifications can prove as an effective method in surgical decompression of quadrangular space syndrome. The authors suggest more cadaveric studies to facilitate anatomists and surgeons with the opportunities to practice and evaluate older and newer surgical approaches.

Keywords: surgical approach, anatomical approach, decompression, axillary nerve, quadrangular space

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Authors: G. Cunningham, E. Cantor, X. Wu, F. Shen, G. Jiang, S. Philips, C. Bales, Y. Xiao, T. R. Cummins, J. C. Fehrenbacher, B. P. Schneider

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Background: Taxane-induced peripheral neuropathy (TIPN) is the most devastating survivorship issue for patients receiving therapy. Dose reductions due to TIPN in the curative setting lead to inferior outcomes for African American patients, as prior research has shown that this group is more susceptible to developing severe neuropathy. The mechanistic underpinnings of TIPN, however, have not been entirely elucidated. While it would be appealing to use primary tissue to study the development of TIPN, procuring nerves from patients is not realistically feasible, as nerve biopsies are painful and may result in permanent damage. Therefore, our laboratory has investigated paclitaxel-induced neuronal morphological and molecular changes using an ex vivo model of human-induced pluripotent stem cell (iPSC)-derived neurons. Methods: iPSCs are undifferentiated and endlessly dividing cells that can be generated from a patient’s somatic cells, such as peripheral blood mononuclear cells (PBMCs). We successfully reprogrammed PBMCs into iPSCs using the Erythroid Progenitor Reprograming Kit (STEMCell Technologiesᵀᴹ); pluripotency was verified by flow cytometry analysis. iPSCs were then induced into neurons using a differentiation protocol that bypasses the neural progenitor stage and uses selected small-molecule modulators of key signaling pathways (SMAD, Notch, FGFR1 inhibition, and Wnt activation). Results: Flow cytometry analysis revealed expression of core pluripotency transcription factors Nanog, Oct3/4 and Sox2 in iPSCs overlaps with commercially purchased pluripotent cell line UCSD064i-20-2. Trilineage differentiation of iPSCs was confirmed with immunofluorescent imaging with germ-layer-specific markers; Sox17 and ExoA2 for ectoderm, Nestin, and Pax6 for mesoderm, and Ncam and Brachyury for endoderm. Sensory neuron markers, β-III tubulin, and Peripherin were applied to stain the cells for the maturity of iPSC-derived neurons. Patch-clamp electrophysiology and calcitonin gene-related peptide (CGRP) release data supported the functionality of the induced neurons and provided insight into the timing for which downstream assays could be performed (week 4 post-induction). We have also performed a cell viability assay and fluorescence-activated cell sorting (FACS) using four cell-surface markers (CD184, CD44, CD15, and CD24) to select a neuronal population. At least 70% of the cells were viable in the isolated neuron population. Conclusion: We have found that these iPSC-derived neurons recapitulate mature neuronal phenotypes and demonstrate functionality. Thus, this represents a patient-derived ex vivo neuronal model to investigate the molecular mechanisms of clinical TIPN.

Keywords: chemotherapy, iPSC-derived neurons, peripheral neuropathy, taxane, paclitaxel

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Authors: S. alireza Mirghasemi, Shervin Rashidinia, Mohammadsaleh Sadeghi, Mohsen Talebizadeh, Narges Rahimi Gabaran, S. Shahin Eftekhari, Sara Shahmoradi

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Purpose: There are various pin-in-plaster methods for treating distal radius fractures. This study is meant to introduce a modified technique of pin-in-plaster. Materials and methods: Fifty-four patients with distal radius fractures were followed up for one year. Patients were excluded if they had type B fractures according to AO classification, multiple injuries or pathological fractures, and were treated more than 7 days after injury. Range of motion and functional results were evaluated. Radiographic parameters including radial inclination, tilt, and height, were measured preoperatively and postoperatively. Results: The average radial tilt was 10.6° and radial height was 10.2 mm at the sixth month postoperatively. Three cases of pin tract infection were recorded, who were treated totally with oral antibiotics. There was no case of pin loosening. Of total 73 patients underwent surgery, three cases of radial nerve irritation were recorded at the time of cast removal. All of them resolved at the 6th month follow up. No median nerve compression and carpal tunnel syndrome have found. We also had no case of tendon injury. Conclusion: Our modified technique is effective to restore anatomic congruity and maintain reduction.

Keywords: distal radius fracture, percutaneous pinning, pin-in-plaster, modified method of pin-in-plaster, operative treatment

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Authors: P. Renjitha, P. Hari Prakash

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Auditory neuropathy spectrum disorder (ANSD) is an auditory disorder with normal cochlear outer hair cell function and disrupted auditory nerve function. It results in unique clinical characteristic with absent auditory brainstem response (ABR), absent acoustic reflex and the presence of otoacoustic emissions (OAE) and cochlear microphonics. The lesion site could be at cochlear inner hair cells, the synapse between the inner hair cells and type I auditory nerve fibers, and/or the auditory nerve itself. But the literatures on synchrony at higher auditory system are sporadic and are less understood. It might be interesting to see if there is a recovery of neural synchrony at higher auditory centers. Also, does the level at which the auditory system recovers with adequate synchrony to the extent of observable evoke response potentials (ERPs) can predict speech perception? In the current study, eight ANSD participants and healthy controls underwent detailed audiological assessment including ABR, auditory middle latency response (AMLR), and auditory late latency response (ALLR). AMLR was recorded for clicks and ALLR was evoked using 500Hz and 2 kHz tone bursts. Analysis revealed that the participant could be categorized into three groups. Group I (2/8) where ALLR was present only for 2kHz tone burst. Group II (4/8), where AMLR was absent and ALLR was seen for both the stimuli. Group III (2/8) consisted individuals with identifiable AMLR and ALLR for all the stimuli. The highest speech identification sore observed in ANSD group was 30% and hence considered having poor speech perception. Overall test result indicates that the site of neural synchrony recovery could be varying across individuals with ANSD. Some individuals show recovery of neural synchrony at the thalamocortical level while others show the same only at the cortical level. Within ALLR itself there could be variation across stimuli again could be related to neural synchrony. Nevertheless, none of these patterns could possible explain the speech perception ability of the individuals. Hence, it could be concluded that neural synchrony as measured by evoked potentials could not be a good clinical predictor speech perception.

Keywords: auditory late latency response, auditory middle latency response, auditory neuropathy spectrum disorder, correlation with speech identification score

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Authors: Z. B. Ibrahim, N. Ismail, K. I. Othman

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Fuzzy Differential Equations (FDEs) play an important role in modelling many real life phenomena. The FDEs are used to model the behaviour of the problems that are subjected to uncertainty, vague or imprecise information that constantly arise in mathematical models in various branches of science and engineering. These uncertainties have to be taken into account in order to obtain a more realistic model and many of these models are often difficult and sometimes impossible to obtain the analytic solutions. Thus, many authors have attempted to extend or modified the existing numerical methods developed for solving Ordinary Differential Equations (ODEs) into fuzzy version in order to suit for solving the FDEs. Therefore, in this paper, we proposed the development of a fuzzy version of three-point block method based on Block Backward Differentiation Formulas (FBBDF) for the numerical solution of first order FDEs. The three-point block FBBDF method are implemented in uniform step size produces three new approximations simultaneously at each integration step using the same back values. Newton iteration of the FBBDF is formulated and the implementation is based on the predictor and corrector formulas in the PECE mode. For greater efficiency of the block method, the coefficients of the FBBDF are stored at the start of the program. The proposed FBBDF is validated through numerical results on some standard problems found in the literature and comparisons are made with the existing fuzzy version of the Modified Simpson and Euler methods in terms of the accuracy of the approximated solutions. The numerical results show that the FBBDF method performs better in terms of accuracy when compared to the Euler method when solving the FDEs.

Keywords: block, backward differentiation formulas, first order, fuzzy differential equations

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Authors: Laila Montaser, Hala Gabr, Maha El-Bassuony, Gehan Tawfeek

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Orthotropic liver transplantation (OLT) is the final procedure of both end stage and metabolic liver diseases. Hepatocyte transplantation is an alternative for OLT, but the sources of hepatocytes are limited. Bone marrow mesenchymal stem cells (BM-MSCs) can differentiate into hepatocyte-like cells and are a potential alternative source for hepatocytes. The MSCs from bone marrow are a promising target population as they are capable of differentiating along multiple lineages and, at least in vitro, have significant expansion capability. MSCs from bone marrow may have the potential to differentiate in vitro and in vivo into hepatocytes. Our study examined whether mesenchymal stem cells (MSCs), which are stem cells originated from human bone marrow, are able to differentiate into functional hepatocyte-like cells in vitro. Our aim was to investigate the differentiation potential of BM-MSCs into hepatocyte-like cells. Adult stem cell therapy could solve the problem of degenerative disorders, including liver disease.

Keywords: bone marrow, differentiation, hepatocyte, stem cells

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Authors: G. Fitzpatrick, S. O. Chonghaile, D. Harmon

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Objective: Chronic pain patients represent a unique challenge in the Emergency Department. Very little literature has been published regarding this group of patients. Our aim was to determine the attitude of patients with chronic pain to the Emergency Department in order to improve and streamline their future visits. Methods: A two-year survey was carried out on Chronic Pain Patients regarding their Emergency Department Attendances. Patients attending the Pain Clinic in Croom Hospital, Co. Limerick were asked to complete a 20-part questionnaire regarding their experiences of visiting the Emergency Department in the preceding year. 46 questionnaires were completed. Results: Unbearable breakthrough pain was the main reason for visiting the Emergency Department. More than half (54%) of those surveyed were not satisfied with the treatment received. Problems indicated included under-treatment of pain (59%), a sense of being under undue suspicion of drug-seeking behaviour (33%) and a perception that the patient themselves understood their condition better than the treating doctor (76%). Paracetamol, NSAIDs, or time off work comprised 72% of the treatments offered – all of which could have been provided by their General Practitioner. Only 4% were offered a nerve block. 67% felt that the creation of personalised Patient Plans, consisting of an agreed plan between the patient, their pain specialist, and the Emergency Department, would expedite their trip through the Emergency Department. Conclusions: Chronic pain patients generally have a negative experience in the ED. Possible future solutions include increasing our empathy and levels of knowledge, provision of nerve blocks in the ED, and use of personalised “Patient Plans” to streamline the treatment pathway for this group of patients.

Keywords: chronic pain, survey, patients, emergency department

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Authors: Hye-Young Shin, Chan Kee Park

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Background: To compare the thinning patterns of the ganglion cell-inner plexiform layer (GCIPL) and peripapillary retinal nerve fiber layer (pRNFL) as measured using Cirrus high-definition optical coherence tomography (HD-OCT) in patients with visual field (VF) defects that respect the vertical meridian. Methods: Twenty eyes of eleven patients with VF defects that respect the vertical meridian were enrolled retrospectively. The thicknesses of the macular GCIPL and pRNFL were measured using Cirrus HD-OCT. The 5% and 1% thinning area index (TAI) was calculated as the proportion of abnormally thin sectors at the 5% and 1% probability level within the area corresponding to the affected VF. The 5% and 1% TAI were compared between the GCIPL and pRNFL measurements. Results: The color-coded GCIPL deviation map showed a characteristic vertical thinning pattern of the GCIPL, which is also seen in the VF of patients with brain lesions. The 5% and 1% TAI were significantly higher in the GCIPL measurements than in the pRNFL measurements (all P < 0.01). Conclusions: Macular GCIPL analysis clearly visualized a characteristic topographic pattern of retinal ganglion cell (RGC) loss in patients with VF defects that respect the vertical meridian, unlike pRNFL measurements. Macular GCIPL measurements provide more valuable information than pRNFL measurements for detecting the loss of RGCs in patients with retrograde degeneration of the optic nerve fibers.

Keywords: brain lesion, macular ganglion cell, inner plexiform layer, spectral-domain optical coherence tomography

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Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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Authors: Waseem Al Talalwah, Shorok Ali Al Dorazi, Roger Soames

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The inferior gluteal artery is a largest branch of the anterior division of internal iliac artery. It escapes from the pelvic cavity through the greater sciatic foramen below the lower edge of piriformis. In gluteal region, it provides several muscular branches to gluteal maximus and articular branch to hip joint. Further, it provides sciatic branch to sciatic nerve. Present study explores the origin of the inferior gluteal artery of 41 cadavers in Centre for Anatomy and Human Identification, University of Dundee, UK. It arose directly from the anterior division of internal iliac artery in 39% and 45.7% indirectly as with the internal pudendal artery. Further, it arose indirectly from anterior division with internal pudendal and obturator arteries in 1.5% referred as obturatogluteopudendal trunk in 1.5%. Therefore, it arose from the anterior division of the internal iliac artery in 86.2%. However, it found to be as a branch of the posterior division of internal iliac artery in 7.7% which is either a direct branch in 6.2% as or indirect branch (as from the sciatic artery) in 1.5%. It neither arose from anterior or posterior division in 1.5% as from gluteopudendal trunk arising from the internal iliac artery bifurcation site. In few cases, the inferior gluteal artery found to be congenital absence in 4.6% which is compensated by the persistent sciatic artery. Therefore, radiologists have to aware of the origin variability of the inferior gluteal artery to alert surgeons. Knowing the origin of the inferior gluteal artery may help the surgeons to avoid iatrogenic sciatic neuropathy or gluteal claudication due to prolonged ligation in pelvic procedures such as removing prostate or of uterine fibroid.

Keywords: inferior gluteal artery, internal pudendal, sciatic nerve, sciatic artery, gluteal claudication, sciatic neuopathy

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Authors: W. Baumgartner, M. Welti, N. Hild, S. C. Hess, W. J. Stark, G. Meier Bürgisser, P. Giovanoli, J. Buschmann

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Perfusion bioreactors are used to solve problems in tissue engineering in terms of sufficient nutrient and oxygen supply. Such problems especially occur in critical size grafts because vascularization is often too slow after implantation ending up in necrotic cores. Biominerizable and biocompatible nanocomposite materials are attractive and suitable scaffold materials for bone tissue engineering because they offer mineral components in organic carriers – mimicking natural bone tissue. In addition, human adipose derived stem cells (ASCs) can potentially be used to increase bone healing as they are capable of differentiating towards osteoblasts or endothelial cells among others. In the present study, electrospun nanocomposite disks of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/a-CaP) were seeded with human ASCs and eight disks were stacked in a bioreactor running with normal culture medium (no differentiation supplements). Under continuous perfusion and uniaxial cyclic compression, load-displacement curves as a function of time were assessed. Stiffness and energy dissipation were recorded. Moreover, stem cell densities in the layers of the piled scaffold were determined as well as their morphologies and differentiation status (endothelial cell differentiation, chondrogenesis and osteogenesis). While the stiffness of the cell free constructs increased over time caused by the transformation of the a-CaP nanoparticles into flake-like apatite, ASC-seeded constructs showed a constant stiffness. Stem cell density gradients were histologically determined with a linear increase in the flow direction from the bottom to the top of the 3.5 mm high pile (r2 > 0.95). Cell morphology was influenced by the flow rate, with stem cells getting more roundish at higher flow rates. Less than 1 % osteogenesis was found upon osteopontin immunostaining at the end of the experiment (9 days), while no endothelial cell differentiation and no chondrogenesis was triggered under these conditions. All ASCs had mainly remained in their original pluripotent status within this time frame. In summary, we have fabricated a critical size bone graft based on a biominerizable bone-biomimetic nanocomposite with preserved stiffness when seeded with human ASCs. The special feature of this bone graft was that ASC densities inside the piled construct varied with a linear gradient, which is a good starting point for tissue engineering interfaces such as bone-cartilage where the bone tissue is cell rich while the cartilage exhibits low cell densities. As such, this tissue-engineered graft may act as a bone-cartilage interface after the corresponding differentiation of the ASCs.

Keywords: bioreactor, bone, cartilage, nanocomposite, stem cell gradient

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Authors: Hoo Cheol Lee, Dae Seung Kim, Sang Myung Jung, Gwang Heum Yoon, Hwa Sung Shin

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Loss of bone tissue can occur due to a bone tissue disease and aging or fracture. Renewable formation of bone is mainly made by its differentiation and metabolism. For this reason, osteoblasts have been studied for regeneration of bone tissue. So, tissue engineering has attracted attention as a recovery means. In tissue engineering, a particularly important factor is a scaffold that supports cell growth. For osteoblast scaffold, we used the cellulose nanowhisker (CNW) extracted from marine organism. CNW is one of an abundant material obtained from a number of plants and animals. CNW is polymer consisting of monomer cellulose and this composition offers biodegradability and biocompatibility to CNW. Mechanical strength of CNW is superior to the existing natural polymers. In addition, substances of marine origin have a low risk of secondary infection by bacteria and pathogen in contrast with those of land-derived. For evaluating its suitability as an osteoblast scaffold, we fabricate CNW film for osteoblast culture and performed the MTT assay and ALP assay to confirm its cytotoxicity and effect on differentiation. Taking together these results, we assessed CNW is a potential candidate of a material for bone tissue regeneration.

Keywords: bone regeneration, cellulose nanowhisker, marine derived material, osteoblast

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Authors: Trevor E. Davis, Isaac Cassar, Yi-Kai Lo, Wentai Liu

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This study models the use of transcutaneous electrical nerve stimulation on skin with a disk electrode in order to simulate tissue damage. The current density distribution above a disk electrode is known to be a dynamic and non-uniform quantity that is intensified at the edges of the disk. The non-uniformity is subject to change through using various electrode geometries or stimulation methods. One of these methods known as edge-retarded stimulation has shown to reduce this edge enhancement. Though progress has been made in modeling the behavior of a disk electrode, little has been done to test the validity of these models in simulating the actual heat transfer from the electrode. This simulation uses finite element software to couple the injection of current from a disk electrode to heat transfer described by the Pennesbioheat transfer equation. An example application of this model is studying an experimental form of stimulation, known as edge-retarded stimulation. The edge-retarded stimulation method will reduce the current density at the edges of the electrode. It is hypothesized that reducing the current density edge enhancement effect will, in turn, reduce temperature change and tissue damage at the edges of these electrodes. This study tests this hypothesis as a demonstration of the capabilities of this model. The edge-retarded stimulation proved to be safer after this simulation. It is shown that temperature change and the fraction of tissue necrosis is much greater in the square wave stimulation. These results bring implications for changes of procedures in transcutaneous electrical nerve stimulation and transcutaneous spinal cord stimulation as well.

Keywords: bioheat transfer, electrode, neuroprosthetics, TENS, transcutaneous stimulation

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Authors: Sahriye Sonmez, Salih Ulger, Mustafa Kaplan, Mustafa Karhan

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The objective of this study was to investigate the effects of external ALAR application on nutrients contents in leaf and node in ‘on (bearing)’ and ‘off (non-bearing)’ years in Memecik olive trees. For this purpose; 2000 mg L-1 ALAR was externally applied to Memecik olive trees, and leaf and node samples from olive trees were taken during the induction, initiation and differentiation periods in ‘on’ and ‘off’ years. Nutrients contents (N, P, K, Ca, Mg, Fe, Mn, Zn and Cu) in leaf and node samples were determined. The K, Ca, Mg, Fe, Mn, Zn and Cu contents were determined by atomic absorption spectrophotometry, Nitrogen by Kjeldahl procedure, and P by a spectrophotometric method. The results showed that the N, Ca, Mg, Fe, Mn, Zn and Cu contents in ‘on’ year were higher than ‘off’ year while the K contents in ‘on’ year were lower than ‘off ‘ year, but the P content was not different. The N, Ca, Mg, Fe and Mn contents in leaf samples were higher in the node samples except for K while the P, Zn and Cu contents were not different. The N, K, Ca, Fe, Mn, Zn and Cu contents were lowest during the initiation period while the P content was highest in this period. The Mg content was not different in all period.

Keywords: bearing, differentiation period, induction period, initiation period, non bearing, olive

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Authors: Yuko Ishiwaka, Tomohiro Yoshida, Tadateru Itoh

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Human makes preparation called readiness potential unconsciously (RP) before awareness of their own decision. For example, when recognizing a button and pressing the button, the RP peaks are observed 200 ms before the initiation of the movement. It has been known that the preparatory movements are acquired before actual movements, but it has not been still well understood how humans can obtain the RP during their growth. On the proposition of why the brain must respond earlier, we assume that humans have to adopt the dangerous environment to survive and then obtain the behavior to cover the various time lags distributed in the body. Without RP, humans cannot take action quickly to avoid dangerous situations. In taking action, the brain makes decisions, and signals are transmitted through the Spinal Cord to the muscles to the body moves according to the laws of physics. Our research focuses on the time lag of the neuron signal transmitting from a brain to muscle via a spinal cord. This time lag is one of the essential factors for readiness potential. We propose a firing pattern controller model of a motor nerve system considered innervation and jitter, which produces time lag. In our simulation, we adopt innervation and jitter in our proposed muscle-skeleton model, because these two factors can create infinitesimal time lag. Q10 Hodgkin Huxley model to calculate action potentials is also adopted because the refractory period produces a more significant time lag for continuous firing. Keeping constant power of muscle requires cooperation firing of motor neurons because a refractory period stifles the continuous firing of a neuron. One more factor in producing time lag is slow or fast-twitch. The Expanded Hill Type model is adopted to calculate power and time lag. We will simulate our model of muscle skeleton model by controlling the firing pattern and discuss the relationship between the time lag of physics and neurons. For our discussion, we analyze the time lag with our simulation for knee bending. The law of inertia caused the most influential time lag. The next most crucial time lag was the time to generate the action potential induced by innervation and jitter. In our simulation, the time lag at the beginning of the knee movement is 202ms to 203.5ms. It means that readiness potential should be prepared more than 200ms before decision making.

Keywords: firing patterns, innervation, jitter, motor nerve system, readiness potential

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Authors: Ana Ferreira, Ines Costa, Patricia Polónia, Josué Pereira, Olinda Faria, Pedro Alberto Silva

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Introduction: Optical coherence tomography (OCT) and visual field tools are pivotal in evaluating neurological deficits and predicting potential visual improvement following surgical decompression in neurosurgical patients. Despite their clinical significance, a comprehensive understanding of their utility in this context is lacking in the literature. This study aims to elucidate the applications of OCT and visual field assessment, delineating distinct patterns of visual deficit presentations within the studied cohort. Methods: This retrospective analysis considered all adult patients who underwent a single surgery for pituitary adenoma or anterior skull base meningioma with optic nerve involvement, coupled with neuro-ophthalmology evaluation, between July 2020 and January 2023. A minimum follow-up period of 6 months was deemed essential. Results: A total of 24 patients, with a median age of 61, were included in the analysis. Three primary patterns emerged: 1) Low visual field involvement with compromised OCT, 2) High visual field involvement with relatively unaffected OCT, and 3) Significant compromise observed in both OCT and visual fields. Conclusion: This study delineates various findings in OCT and visual field assessments with illustrative examples. Based on the current findings, a prospective cohort will be systematically collected to further investigate and validate these patterns and their prognostic significance, enhancing our understanding of the utility of OCT and visual fields in neurosurgical patients.

Keywords: OCT, neurosurgery, visual field, optic nerve

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Authors: Tae-Hoon Kim, Kwon-Ha Yoon, Hong Young Jun, Ki-Jong Kim, Young Hwan Lee, Myeung Su Lee, Keum Ha Choi, Ki Jung Yun, Eun Young Cho, Yong-Yeon Jeong, Chung-Hwan Jun

Abstract:

Purpose: To assess the changes of hepatic metabolite for differentiation between non-alcoholic steatohepatitis (NASH) and simple steatosis on proton magnetic resonance spectroscopy (1H-MRS) in both humans and animal model. Methods: The local institutional review board approved this study and subjects gave written informed consent. 1H-MRS measurements were performed on a localized voxel of the liver using a point-resolved spectroscopy (PRESS) sequence and hepatic metabolites of alanine (Ala), lactate/triglyceride (Lac/TG), and TG were analyzed in NASH, simple steatosis and control groups. The group difference was tested with the ANOVA and Tukey’s post-hoc tests, and diagnostic accuracy was tested by calculating the area under the receiver operating characteristics (ROC) curve. The associations between metabolic concentration and pathologic grades or non-alcoholic fatty liver disease(NAFLD) activity scores were assessed by the Pearson’s correlation. Results: Patient with NASH showed the elevated Ala(p<0.001), Lac/TG(p < 0.001), TG(p < 0.05) concentration when compared with patients who had simple steatosis and healthy controls. The NASH patients were higher levels in Ala(mean±SEM, 52.5±8.3 vs 2.0±0.9; p < 0.001), Lac/TG(824.0±168.2 vs 394.1±89.8; p < 0.05) than simple steatosis. The area under the ROC curve to distinguish NASH from simple steatosis was 1.00 (95% confidence interval; 1.00, 1.00) with Ala and 0.782 (95% confidence interval; 0.61, 0.96) with Lac/TG. The Ala and Lac/TG levels were well correlated with steatosis grade, lobular inflammation, and NAFLD activity scores. The metabolic changes in human were reproducible to a mice model induced by streptozotocin injection and a high-fat diet. Conclusion: 1H-MRS would be useful for differentiation of patients with NASH and simple hepatic steatosis.

Keywords: non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, 1H MR spectroscopy, hepatic metabolites

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Authors: Xuan Zhao, Wen Jiang

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In this study, a web crawler was used to collect POI sample data from 38 districts and counties of Chongqing in 2022, and ArcGIS was combined to coordinate and projection conversion and realize data visualization. Nuclear density analysis and spatial correlation analysis were used to explore the spatial distribution characteristics of elderly care facilities in Chongqing, and K mean cluster analysis was carried out with GeoDa to study the spatial concentration degree of elderly care resources in 38 districts and counties. Finally, the driving force of spatial differentiation of elderly care facilities in various districts and counties of Chongqing is studied by using the method of geographic detector. The results show that: (1) in terms of spatial distribution structure, the distribution of elderly care facilities in Chongqing is unbalanced, showing a distribution pattern of ‘large dispersion and small agglomeration’ and the asymmetric pattern of ‘west dense and east sparse, north dense and south sparse’ is prominent. (2) In terms of the spatial matching between elderly care resources and the elderly population, there is a weak coordination between the input of elderly care resources and the distribution of the elderly population at the county level in Chongqing. (3) The analysis of the results of the geographical detector shows that the single factor influence is mainly the number of elderly population, public financial revenue and district and county GDP. The high single factor influence is mainly caused by the elderly population, public financial income, and district and county GDP. The influence of each influence factor on the spatial distribution of elderly care facilities is not simply superimposed but has a nonlinear enhancement effect or double factor enhancement. It is necessary to strengthen the synergistic effect of two factors and promote the synergistic effect of multiple factors.

Keywords: aging, elderly care facilities, spatial differentiation, geographical detector, driving force analysis, Mountain city

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Authors: Dmitry V. Pasynkov, Ivan A. Egoshin, Alexey A. Kolchev, Ivan V. Kliouchkin

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In most cases, typical cysts are easily recognized at ultrasonography. The specificity of this method for typical cysts reaches 98%, and it is usually considered as gold standard for typical cyst diagnosis. However, it is necessary to have all the following features to conclude the typical cyst: clear margin, the absence of internal echoes and dorsal acoustic enhancement. At the same time, not every breast cyst is typical. It is especially characteristic for protein-contained cysts that may have significant internal echoes. On the other hand, some solid lesions (predominantly malignant) may have cystic appearance and may be falsely accepted as cysts. Therefore we tried to develop the automatic method of cystic and solid breast lesions differentiation. Materials and methods. The input data were the ultrasonography digital images with the 256-gradations of gray color (Medison SA8000SE, Siemens X150, Esaote MyLab C). Identification of the lesion on these images was performed in two steps. On the first one, the region of interest (or contour of lesion) was searched and selected. Selection of such region is carried out using the sigmoid filter where the threshold is calculated according to the empirical distribution function of the image brightness and, if necessary, it was corrected according to the average brightness of the image points which have the highest gradient of brightness. At the second step, the identification of the selected region to one of lesion groups by its statistical characteristics of brightness distribution was made. The following characteristics were used: entropy, coefficients of the linear and polynomial regression, quantiles of different orders, an average gradient of brightness, etc. For determination of decisive criterion of belonging to one of lesion groups (cystic or solid) the training set of these characteristics of brightness distribution separately for benign and malignant lesions were received. To test our approach we used a set of 217 ultrasonic images of 107 cystic (including 53 atypical, difficult for bare eye differentiation) and 110 solid lesions. All lesions were cytologically and/or histologically confirmed. Visual identification was performed by trained specialist in breast ultrasonography. Results. Our system correctly distinguished all (107, 100%) typical cysts, 107 of 110 (97.3%) solid lesions and 50 of 53 (94.3%) atypical cysts. On the contrary, with the bare eye it was possible to identify correctly all (107, 100%) typical cysts, 96 of 110 (87.3%) solid lesions and 32 of 53 (60.4%) atypical cysts. Conclusion. Automatic approach significantly surpasses the visual assessment performed by trained specialist. The difference is especially large for atypical cysts and hypoechoic solid lesions with the clear margin. This data may have a clinical significance.

Keywords: breast cyst, breast solid lesion, differentiation, ultrasonography

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Authors: Ankush M. Dewle, Suditi Bhattacharya, Prachi R. Abhang, Savita Datar, Ajay J. Jog, Rupesh K. Srivastava, Geetanjali Tomar

Abstract:

Objectives: The aim of this study was to investigate the quality of mesenchymal stem cells (MSCs) obtained from ageing gingival tissues, in order to suggest their potential role in autologous stem cell therapy for old individuals. Methods: MSCs were isolated from gingival tissues of young (18-45 years) and old (above 45 years) donors by enzymatic digestion. MSCs were analysed for cfu-f, surface marker expression by flow-cytometry and multilineage differentiation potential. The angiogenic potential was compared in a chick embryo yolk sac membrane model. The aging and differentiation markers including SA-β-galactosidase and p21 respectively were analysed by staining and flow-cytometry analysis. Additionally, osteogenic markers such as glucocorticoid receptor (GR), vitamin D receptor (VDR) were measured by flow-cytometry and RT-qPCR was performed for quantification of osteogenic gene expression. Alizarin Red S and alkaline phosphatase (ALP) activity were also quantitated. Results: Gingival MSCs (GMSCs) from both the age groups were similar in their morphology and displayed cfu-f. They had similar expression of MSC surface markers and p21, comparable rate of proliferation and differentiated to all the four lineages. GMSCs from young donors had a higher adipogenic differentiation potential as compared to the old GMSCs. Moreover, these cells did not display a significant difference in ALP activity probably due to comparable expression of GR, VDR, and osteogenic genes. Conclusions: Ageing of GMSCs occurs at a much slower rate than stem cells from other sources. Thus we suggest GMSCs as an excellent candidate for autologous stem cell therapy in degenerative diseases of elderly individuals. Clinical Significance: GMSCs could help overcome the setbacks in clinical implementation of autologous stem cell therapy for regenerative medicine in all age group of patient.

Keywords: bone regeneration, cell therapy, senescence, stem cell

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Authors: Andreas Aceranti, Sergio Romanò, Simonetta Vernocchi, Silvia Arnaboldi, Emilio Mazza

Abstract:

As a result of the Sars-Cov2 pandemic, the incidence of thymism disorders has significantly increased and, often, patients are reluctant to want to take drugs aimed at stabilizing mood. In order to provide an alternative approach to drug therapies, we have prepared a study in order to evaluate the possibility of improving the quality of life of these subjects through osteopathic treatment. Patients were divided into visceral and fascial manual treatment with the aim of increasing serotonin levels and stimulating the vagus nerve through validated techniques. The results were evaluated through the administration of targeted questionnaires in order to assess quality of life, mood, sleep and intestinal functioning. At a first endpoint we found, in patients undergoing fascial treatment, an increase in quality of life and sleep: in fact, they report a decrease in the number of nocturnal awakenings; a reduction in falling asleep times and greater rest upon waking. In contrast, patients undergoing visceral treatment, as well as those included in the control group, did not show significant improvements. Patients in the fascial group have, in fact, reported an improvement in thymism and subjective quality of life with a generalized improvement in function. Although the study is still ongoing, based on the results of the first endpoint we can hypothesize that fascial stimulation of the vagus nerve with manual and osteopathic techniques may be a valid alternative to pharmacological treatments in mood and sleep disorders.

Keywords: ostheopathy, insomnia, noctural awakening, thymism

Procedia PDF Downloads 90

Authors: P. Siriarchawatana, K. Leungchavaphongse, N. Covavisaruch, K. Rojananuangnit, P. Boondaeng, N. Panyayingyong

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Glaucoma is a disease that causes visual loss in adults. Glaucoma causes damage to the optic nerve and its overall pathophysiology is still not fully understood. Vasculopathy may be one of the possible causes of nerve damage. Photographic imaging of retinal vessels by fundus camera during eye examination may complement clinical management. This paper presents an innovation for measuring central light reflex width-to-retinal vessel diameter ratio (CRR) from digital retinal photographs. Using our edge detection technique, CRRs from glaucoma and normal eyes were compared to examine differences and associations. CRRs were evaluated on fundus photographs of participants from Mettapracharak (Wat Raikhing) Hospital in Nakhon Pathom, Thailand. Fifty-five photographs from normal eyes and twenty-one photographs from glaucoma eyes were included. Participants with hypertension were excluded. In each photograph, CRRs from four retinal vessels, including arteries and veins in the inferotemporal and superotemporal regions, were quantified using edge detection technique. From our finding, mean CRRs of all four retinal arteries and veins were significantly higher in persons with glaucoma than in those without glaucoma (0.34 vs. 0.32, p < 0.05 for inferotemporal vein, 0.33 vs. 0.30, p < 0.01 for inferotemporal artery, 0.34 vs. 0.31, p < 0.01 for superotemporal vein, and 0.33 vs. 0.30, p < 0.05 for superotemporal artery). From these results, an increase in CRRs of retinal vessels, as quantitatively measured from fundus photographs, could be associated with glaucoma.

Keywords: glaucoma, retinal vessel, central light reflex, image processing, fundus photograph, edge detection

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Authors: Maki Mizogami, Hironori Tsuchiya, Yoshiroh Hayabuchi, Kenji Shigemi

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Since drugs exhibit significant structure-dependent differences in activity and toxicity, their differentiation based on the mechanism of action should have implications for comparative drug efficacy and safety. We aimed to differentiate drug stereoisomers by their stereostructure-selective membrane interactions underlying pharmacological and toxicological effects. Biomimetic lipid bilayer membranes were prepared with phospholipids and sterols (either cholesterol or epicholesterol) to mimic the lipid compositions of neuronal and cardiomyocyte membranes and to provide these membranes with the chirality. The membrane preparations were treated with different classes of stereoisomers at clinically- and pharmacologically-relevant concentrations (25-200 μM), followed by measuring fluorescence polarization to determine the membrane interactivity of drugs to change the physicochemical property of membranes. All the tested drugs acted on lipid bilayers to increase or decrease the membrane fluidity. Drug stereoisomers could not be differentiated when interacting with the membranes consisting of phospholipids alone. However, they stereostructure-selectively interacted with neuro-mimetic and cardio-mimetic membranes containing 40 mol% cholesterol ((3β)-cholest-5-en-3-ol) to show the relative potencies being local anesthetic R(+)-bupivacaine > rac-bupivacaine > S(‒)-bupivacaine, α2-adrenergic agonistic D-medetomidine > rac-medetomidine > L-medetomidine, β-adrenergic antagonistic R(+)-propranolol > rac-propranolol > S(–)-propranolol, NMDA receptor antagonistic S(+)-ketamine > rac-ketamine, analgesic monoterpenoid (+)-menthol > (‒)-menthol, non-steroidal anti-inflammatory S(+)-ibuprofen > rac-ibuprofen > R(‒)-ibuprofen, and bioactive flavonoid (+)-epicatechin > (‒)-epicatechin. All of the order of membrane interactivity were correlated to those of beneficial and adverse effects of the tested stereoisomers. In contrast, the membranes prepared with epicholesterol ((3α)-chotest-5-en-3-ol), an epimeric form of cholesterol, reversed the rank order of membrane interactivity to be S(‒)-enantiomeric > racemic > R(+)-enantiomeric bupivacaine, L-enantiomeric > racemic > D-enantiomeric medetomidine, S(–)-enantiomeric > racemic > R(+)-enantiomeric propranolol, racemic > S(+)-enantiomeric ketamine, (‒)-enantiomeric > (+)-enantiomeric menthol, R(‒)-enantiomeric > racemic > S(+)-enantiomeric ibuprofen, and (‒)-enantiomeric > (+)-enantiomeric epicatechin. The opposite configuration allows drug molecules to interact with chiral sterol membranes enantiomer-selectively. From the comparative results, it is speculated that a 3β-hydroxyl group in cholesterol is responsible for the enantioselective interactions of drugs. In conclusion, the differentiation of drug stereoisomers by their stereostructure-selective membrane interactions would be useful for designing and predicting drugs with higher activity and/or lower toxicity.

Keywords: chiral membrane, differentiation, drug stereoisomer, enantioselective membrane interaction

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