Search results for: temporal bone carcinoma

Authors: Badawia Ibrahim, Iman Hussein, Samar El Sheikh, Fatma Abou Elkasem, Hazem Abo Ismael

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Background: Response of breast carcinoma to neoadjuvant chemotherapy (NAC) varies regarding many factors including hormonal receptor status. Breast cancer is a heterogenous disease with different outcomes, hence a need arises for new markers predicting the outcome of NAC especially for the triple negative group when estrogen, progesterone receptors and Her2/neu are negative. FOXP3 is a promising target with unclear role. Aim: To examine the value of FOXP3 expression in locally advanced triple negative breast cancer tumoral cells as well as tumor infiltrating lymphocytes (TILs) and to elucidate its relation to the extent of NAC response. Material and Methods: Forty five cases of immunohistochemically confirmed to be triple negative breast carcinoma were evaluated for NAC (Doxorubicin, Cyclophosphamide AC x 4 cycles + Paclitaxel x 12 weeks, patients with ejection fraction less than 60% received Taxotere or Cyclophosphamide, Methotrexate, Fluorouracil CMF) response in both tumour and lymph nodes status according to Miller & Payne's and Sataloff's systems. FOXP3 expression in tumor as well as TILs evaluated in the pretherapy biopsies was correlated with NAC response in breast tumor and lymph nodes as well as other clinicopathological factors. Results: Breast tumour cells showed FOXP3 positive cytoplasmic expression in (42%) of cases. High FOXP3 expression percentage was detected in (47%) of cases. High infiltration by FOXP3+TILs was detected in (49%) of cases. Positive FOXP3 expression was associated with negative lymph node metastasis. High FOXP3 expression percentage and high infiltration by FOXP3+TILs were significantly associated with complete therapy response in axillary lymph nodes. High FOXP3 expression in tumour cells was associated with high infiltration by FOXP3+TILs. Conclusion: This result may provide evidence that FOXP3 marker is a good prognostic and predictive marker for triple negative breast cancer (TNBC) indicated for neoadjuvant chemotherapy and can be used for stratifications of TNBC cases indicated for NAC. As well, this study confirmed the fact that the tumour cells and the surrounding microenvironment interact with each other and the tumour microenvironment can influence the treatment outcomes of TNBC.

Keywords: breast cancer, FOXP3 expression, prediction of neoadjuvant chemotherapy effect, triple negative

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Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson

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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.

Keywords: cell migration, hMSCs, SiHA, transwell migration system

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Authors: Isalira Peroba Ramos, Cherley Borba Vieira de Andrade, Grazielle Suhett, Camila Salata, Paulo Cesar Canary, Guilherme Visconde Brasil, Antonio Carlos Campos de Carvalho, Regina Coeli dos Santos Goldenberg

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Background: The therapeutic options for patients with cancer now include increasingly complex combinations of medications, radiation therapy (RT), and surgical intervention. Many of these treatments have important potential adverse cardiac effects and are likely to have significant effects on patient outcomes. Cell therapy appears to be promising for the treatment of chronic and degenerative diseases, including cardiomyopathy induced by RT, as the current therapeutic options are insufficient. Aims: To evaluate the potential of bone marrow mesenchymal cells (BMMCs) in radioinduced cardiac damage Methods: Female Wistar rats, 3 months old (Ethics Committee 054/14), were divided into 2 groups, non-treated irradiated group (IR n=15) and irradiated and BMMC treated (IRT n=10). Echocardiography was performed to evaluate heart function. After euthanasia, 3 months post treatment; the left ventricle was removed and prepared for RT-qPCR (VEGF and Pro Collagen I) and histological (picrosirius) analysis. Results: In both groups, 45 days after irradiation, ejection fraction (EF) was in the normal range for these animals (> 70%). However, the BMMC treated group had EF (83.1%±2.6) while the non-treated IR group showed a significant reduction (76.1%±2.6) in relation to the treated group. In addition, we observed an increase in VEGF gene expression and a decrease in Pro Collagen I in IRT when compared to IR group. We also observed by histology that the collagen deposition was reduced in IRT (10.26%±0.83) when compared to IR group (25.29%±0.96). Conclusions: Treatment with BMMCs was able to prevent ejection fraction reduction and collagen deposition in irradiated animals. The increase of VEGF and the decrease of pro collagen I gene expression might explain, at least in part, the cell therapy benefits. All authors disclose no financial or personal relationships with individuals or organizations that could be perceived to bias their work. Sources of funding: FAPERJ, CAPES, CNPq, MCT.

Keywords: mesenchymal cells, radioation, cardiotoxicity, bone marrow

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Authors: Gannouni Sonia, Hammami Asma, Saidi Salwa, Rebai Noamen

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Water resources in Tunisia have experienced quantitative and qualitative degradation, especially when talking about wetlands and Sbekhas. Indeed, the objective of this work is to study the spatio-temporal evolution of salinity for 29 years (from 1987 to 2016). A study of the connection between surface water and groundwater is necessary to know the degree of influence of the Sebkha brines on the water table. The evolution of surface salinity is determined by remote sensing based on Landsat TM and OLI/TIRS satellite images of the years 1987, 2007, 2010, and 2016. The processing of these images allowed us to determine the NDVI(Normalized Difference Vegetation Index), the salinity index, and the surface temperature around Sebkha. In addition, through a geographic information system(GIS), we could establish a map of the distribution of salinity in the subsurface of the water table of Chott Mariem and Hergla/SidiBouAli/Kondar. The results of image processing and the calculation of the index and surface temperature show an increase in salinity downstream of in addition to the sebkha and the development of vegetation cover upstream and the western part of the sebkha. This richness may be due both to contamination by seawater infiltration from the barrier beach of Hergla as well as the passage of groundwater to the sebkha.

Keywords: spatio-temporal monitoring, salinity, satellite images, NDVI, sebkha

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Authors: Sanjay Kumar Srivastava, Anu Rani Sharma, Kamna Sachdeva

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Every year, fog formation over the Indo-Gangetic Plains (IGPs) of Indian region during the winter months of December and January is believed to create numerous hazards, inconvenience, and economic loss to the inhabitants of this densely populated region of Indian subcontinent. The aim of the paper is to analyze the spatial and temporal variability of winter fog over IGPs. Long term ground observations of visibility and other meteorological parameters (1971-2010) have been analyzed to understand the formation of fog phenomena and its relevance during the peak winter months of January and December over IGP of India. In order to examine the temporal variability, time series and trend analysis were carried out by using the Mann-Kendall Statistical test. Trend analysis performed by using the Mann-Kendall test, accepts the alternate hypothesis with 95% confidence level indicating that there exists a trend. Kendall tau’s statistics showed that there exists a positive correlation between time series and fog frequency. Further, the Theil and Sen’s median slope estimate showed that the magnitude of trend is positive. Magnitude is higher during January compared to December for the entire IGP except in December when it is high over the western IGP. Decade wise time series analysis revealed that there has been continuous increase in fog days. The net overall increase of 99 % was observed over IGP in last four decades. Diurnal variability and average daily persistence were computed by using descriptive statistical techniques. Geo-statistical analysis of fog was carried out to understand the spatial variability of fog. Geo-statistical analysis of fog revealed that IGP is a high fog prone zone with fog occurrence frequency of more than 66% days during the study period. Diurnal variability indicates the peak occurrence of fog is between 06:00 and 10:00 local time and average daily fog persistence extends to 5 to 7 hours during the peak winter season. The results would offer a new perspective to take proactive measures in reducing the irreparable damage that could be caused due to changing trends of fog.

Keywords: fog, climatology, Mann-Kendall test, trend analysis, spatial variability, temporal variability, visibility

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Authors: C. Gallottini, W. Di Mari, A. Amaddeo, K. Barbaro, A. Dolci, G. Dolci, L. Gallottini, G. Barraco, S. Eramo

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A group of 10 dogs (group A) with Periodontal Disease in the third stage, were subjected to regenerative therapy of periodontal tissues, by use of nano hydroxy apatite (NHA). These animals induced by general anesthesia, where treated by ultrasonic scaling, root planning, and at the end by a mucogingival flap in which it was applied NHA. The flap was closed and sutured with simple steps. Another group of 10 dogs (group B), control group, was treated only by scaling and root planning. No patient was subjected to antibiotic therapy. After three months, a check was made by inspection of the oral cavity, radiography and bone biopsy at the alveolar level. Group A showed a total restitutio ad integrum of the periodontal structures, and in group B still mild gingivitis in 70% of cases and 30% of the state remains unchanged. Numerous experimental studies both in animals and humans have documented that the grafts of porous hydroxyapatite are rapidly invaded by fibrovascular tissue which is subsequently converted into mature lamellar bone tissue by activating osteoblast. Since we acted on the removal of necrotic cementum and rehabilitating the root tissue by polishing without intervention in the ligament but only on anatomical functional interface of cement-blasts, we can connect the positive evolution of the clinical-only component of the cement that could represent this perspective, the only reason that Periodontal Disease become a Cement Disease, while all other clinical elements as nothing more than a clinical pathological accompanying.

Keywords: nanoidroxiaphatite, parodontal disease, cement disease, regenerative therapy

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Authors: V. Schmidt, L. Janovák, N. Wiegand, B. Patczai, K. Turzó

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Large bone defects are not able to heal spontaneously. Bioactive glasses seem to be appropriate (bio)materials for bone reconstruction. Bioactive glasses are osteoconductive and osteoinductive, therefore, play a useful role in bony regeneration and repair. Because of their not optimal mechanical properties (e.g., brittleness, low bending strength, and fracture toughness), their applications are limited. Bioactive glass can be used as a coating material applied on metal surfaces. In this way -when using them as implants- the excellent mechanical properties of metals and the biocompatibility and bioactivity of glasses will be utilized. Furthermore, ion release effects of bioactive glasses regarding osteogenic and angiogenic responses have been shown. Silicate bioactive glasses (45S5 Bioglass) induce the release and exchange of soluble Si, Ca, P, and Na ions on the material surface. This will lead to special cellular responses inducing bone formation, which is favorable in the biointegration of the orthopedic prosthesis. The incorporation of other additional elements in the silicate network such as fluorine, magnesium, iron, silver, potassium, or zinc has been shown, as the local delivery of these ions is able to enhance specific cell functions. Although hip and knee prostheses present a high success rate, bacterial infections -mainly implant associated- are serious and frequent complications. Infection can also develop after implantation of hip prostheses, the elimination of which means more surgeries for the patient and additional costs for the clinic. Prosthesis-related infection is a severe complication of orthopedic surgery, which often causes prolonged illness, pain, and functional loss. While international efforts are made to reduce the risk of these infections, orthopedic surgical infections (SSIs) continue to occur in high numbers. It is currently estimated that up to 2.5% of primary hip and knee surgeries and up to 20% of revision arthroplasties are complicated by periprosthetic joint infection (PJIs). According to some authors, these numbers are underestimated, and they are also increasing. Staphylococcus aureus is the leading cause of both SSIs and PJIs, and the prevalence of methicillin-resistant S. aureus (MRSA) is on the rise, particularly in the United States. These deep infections lead to implant removal and consequently increase morbidity and mortality. The study targets this clinical problem using our experience so far with the Ag-doped polymer coatings on Titanium implants. Non-modified or modified (e.g., doped with antibacterial agents, like Ag) bioactive glasses could play a role in the prevention of infections or the therapy of infected tissues. Bioactive glasses have excellent biocompatibility, proved by in vitro cell culture studies of human osteoblast-like MG-63 cells. Ag-doped bioactive glass-scaffold has a good antibacterial ability against Escherichia coli and other bacteria. It may be concluded that these scaffolds have great potential in the prevention and therapy of implant-associated bone infection.

Keywords: antibacterial agents, bioactive glass, hip and knee prosthesis, medical implants

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Authors: Vladimir A. Todiras

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Spatio-temporal models provide new possibilities for real-time action in pest risk analysis. It should be noted that estimation of the possibility and probability of introduction of a pest and of its economic consequences involves many uncertainties. We present a new mapping technique that assesses pest invasion risk using online BioClass software. BioClass is a GIS tool designed to solve multiple-criteria classification and optimization problems based on fuzzy logic and level set methods. This research describes a method for predicting the potential establishment and spread of a plant pest into new areas using a case study: corn rootworm (Diabrotica spp.), tomato leaf miner (Tuta absoluta) and plum fruit moth (Grapholita funebrana). Our study demonstrated that in BioClass we can combine fuzzy logic and geographic information systems with knowledge of pest biology and environmental data to derive new information for decision making. Pests are sensitive to a warming climate, as temperature greatly affects their survival and reproductive rate and capacity. Changes have been observed in the distribution, frequency and severity of outbreaks of Helicoverpa armigera on tomato. BioClass has demonstrated to be a powerful tool for applying dynamic models and map the potential future distribution of a species, enable resource to make decisions about dangerous and invasive species management and control.

Keywords: classification, model, pest, risk

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Authors: Shreya Srivastava, Sushovan Ghosh, Sagnik Dey

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Aerosols directly affect Earth’s radiation budget by scattering and absorbing incoming solar radiation and outgoing terrestrial radiation. While the uncertainty in aerosol radiative forcing (ARF) has decreased over the years, it is still higher than that of greenhouse gas forcing, particularly in the South Asian region, due to high heterogeneity in their chemical properties. Understanding the Spatio-temporal heterogeneity of aerosol composition is critical in improving climate prediction. Studies using satellite data, in-situ and aircraft measurements, and models have investigated the Spatio-temporal variability of aerosol characteristics. In this study, we have taken aerosol data from Multi-angle Imaging Spectro-Radiometer (MISR) level-2 version 23 aerosol products retrieved at 4.4 km and radiation data from Clouds and the Earth’s Radiant Energy System (CERES, spatial resolution=1ox1o) for 21 years (2000-2021) over the Indian subcontinent. MISR aerosol product includes size and shapes segregated aerosol optical depth (AOD), Angstrom exponent (AE), and single scattering albedo (SSA). Additionally, 74 aerosol mixtures are included in version 23 data that is used for aerosol speciation. We have seasonally mapped aerosol optical and microphysical properties from MISR for India at quarter degrees resolution. Results show strong Spatio-temporal variability, with a constant higher value of AOD for the Indo-Gangetic Plain (IGP). The contribution of small-size particles is higher throughout the year, spatially during winter months. SSA is found to be overestimated where absorbing particles are present. The climatological map of short wave (SW) ARF at the top of the atmosphere (TOA) shows a strong cooling except in only a few places (values ranging from +2.5o to -22.5o). Cooling due to aerosols is higher in the absence of clouds. Higher negative values of ARF are found over the IGP region, given the high aerosol concentration above the region. Surface ARF values are everywhere negative for our study domain, with higher values in clear conditions. The results strongly correlate with AOD from MISR and ARF from CERES.

Keywords: aerosol Radiative forcing (ARF), aerosol composition, single scattering albedo (SSA), CERES

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Authors: Zhou Yang, Heli Sun, Jianbin Huang, Jizhong Zhao, Shaojie Qiao

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Traffic prediction is essential in a multitude of ways in modern urban life. The researchers of earlier work in this domain carry out the investigation chiefly with two major focuses: (1) the accurate forecast of future values in multiple time series and (2) knowledge extraction from spatial-temporal correlations. However, two key considerations for traffic prediction are often missed: the completeness of raw data and the full context of the prediction timestamp. Concentrating on the two drawbacks of earlier work, we devise an approach that can address these issues in a two-phase framework. First, we utilize the raw trajectories to a greater extent through building a VLA table and data compression. We obtain the intra-trajectory features with graph-based encoding and the intertrajectory ones with a grid-based model and the technique of back projection that restore their surrounding high-resolution spatial-temporal environment. To the best of our knowledge, we are the first to study direct feature extraction from raw trajectories for traffic prediction and attempt the use of raw data with the least degree of reduction. In the prediction phase, we provide a broader context for the prediction timestamp by taking into account the information that are around it in the training dataset. Extensive experiments on several well-known datasets have verified the effectiveness of our solution that combines the strength of raw trajectory data and prediction context. In terms of performance, our approach surpasses several state-of-the-art methods for traffic prediction.

Keywords: traffic prediction, raw data utilization, context building, data reduction

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Authors: Mark T. Hanson, Alan T. Varughese

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Osteoporosis is a disease affecting bone quality which in turn can increase the risk of low energy fractures. Treatment of osteoporosis using Bisphosphonates has the beneficial effect of increasing bone mass while at the same time has been linked to the formation of atypical femoral fractures. This has led to the increased study of micro-fractures in bones of patients using Bisphosphonate treatment. One of the mechanics related issues which have been identified in this regard is the loss in stiffness of bones containing one or many micro-fractures. Different theories have been put forth using fracture mechanics to determine the effect of crack presence on elastic properties such as modulus. However, validation of these results in a deterministic way has not been forthcoming. The present analysis seeks to provide this deterministic evaluation of fracture’s effect on the elastic modulus. In particular, the effect of crack size, crack orientation and crack number on elastic modulus is investigated. In particular, the Finite Element method is used to explicitly determine the elastic modulus reduction caused by the presence of cracks in a representative volume element. Single cracks of various lengths and orientations are examined as well as cases of multiple cracks. Cracks in tension as well as under shear stress are considered. Although the focus is predominantly two-dimensional, some three-dimensional results are also presented. The results obtained show the explicit reduction in modulus caused by the parameters of crack size, orientation and number noted above. The present results allow the interpretation of the various theories which currently exist in the literature.

Keywords: cracks, elastic, fracture, modulus

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Authors: A. Chari, A. El Bouari, B. Orayech, A. Faik, J. M. Igartua

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The phosphate is a natural resource of great importance in Morocco. In order to exploit this wealth, synthesis and studies of new a material based phosphate, were carried out. The apatite structure present o lot of characteristics, One of the main characteristics is to allow large and various substitutions for both cations and anions. Beside their biological importance in hard tissue (bone and teeth), apatites have been extensively studied for their potential use as fluorescent lamp phosphors or laser host materials.The apatite have interesting possible application fields such as in medicine as materials of bone filling, coating of dental implants, agro chemicals as artificial fertilizers. The LiPb2Ca2(PO4)3 was synthesized by the solid-state method, its crystal structure was investigated by Rietveld analysis using XRPD data. This material crystallizes with a structure of lacunar apatite anion deficit. The LiPb2Ca2(PO4)3 is hexagonal apatite at room temperature, adopting the space group P63/m (ITA No. 176), Rietveld refinements showed that the site 4f is shared by three cations Ca, Pb and Li. While the 6h is occupied by the Pb and Li cations. The structure can be described as built up from the PO4 tetrahedra and the sixfold coordination cavities, which delimit hexagonal tunnels along the c-axis direction. These tunnels are linked by the cations occupying the 4 f sites. Raman and Infrared spectroscopy analyses were carried out. The observed frequencies were assigned and discussed on the basis of unit-cell group analysis and by comparison to other apatite-type materials.

Keywords: apatite, Lacunar, crystal structure, Rietveldmethod, LiPb2Ca2(PO4)3, Phase transition

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Authors: Ashish Jaiswal, Ashwin Ramesh Babu, Mohammad Zaki Zadeh, Fillia Makedon, Glenn Wylie

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Functional magnetic resonance imaging (fMRI) is a neuroimaging technique that records neural activations in the brain by capturing the blood oxygen level in different regions based on the task performed by a subject. Given fMRI data, the problem of predicting the state of cognitive fatigue in a person has not been investigated to its full extent. This paper proposes tackling this issue as a multi-class classification problem by dividing the state of cognitive fatigue into six different levels, ranging from no-fatigue to extreme fatigue conditions. We built a spatio-temporal model that uses convolutional neural networks (CNN) for spatial feature extraction and a long short-term memory (LSTM) network for temporal modeling of 4D fMRI scans. We also applied a self-supervised method called MoCo (Momentum Contrast) to pre-train our model on a public dataset BOLD5000 and fine-tuned it on our labeled dataset to predict cognitive fatigue. Our novel dataset contains fMRI scans from Traumatic Brain Injury (TBI) patients and healthy controls (HCs) while performing a series of N-back cognitive tasks. This method establishes a state-of-the-art technique to analyze cognitive fatigue from fMRI data and beats previous approaches to solve this problem.

Keywords: fMRI, brain imaging, deep learning, self-supervised learning, contrastive learning, cognitive fatigue

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Authors: Emma R. Arakelova, Stepan G. Grigoryan, Flora G. Arsenyan, Nelli S. Babayan, Ruzanna M. Grigoryan, Natalia K. Sarkisyan

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Novel nanosize zinc oxide composites of doxorubicin obtained by deposition of 180 nm thick zinc oxide film on the drug surface using DC-magnetron sputtering of a zinc target in the form of gels (PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO) were studied for drug delivery applications. The cancer specificity was revealed both in in vitro and in vivo models. The cytotoxicity of the test compounds was analyzed against human cancer (HeLa) and normal (MRC5) cell lines using MTT colorimetric cell viability assay. IC50 values were determined and compared to reveal the cancer specificity of the test samples. The mechanistic study of the most active compound was investigated using Flow cytometry analyzing of the DNA content after PI (propidium iodide) staining. Data were analyzed with Tree Star FlowJo software using cell cycle analysis Dean-Jett-Fox module. The in vivo anticancer activity estimation experiments were carried out on mice with inoculated ascitic Ehrlich’s carcinoma at intraperitoneal introduction of doxorubicin and its zinc oxide compositions. It was shown that the nanosize zinc oxide film deposition on the drug surface leads to the selective anticancer activity of composites at the cellular level with the range of selectivity index (SI) from 4 (Starch+NaCMC+Dox+ZnO) to 200 (PEO(gel)+Dox+ZnO) which is higher than that of free Dox (SI = 56). The significant increase in vivo antitumor activity (by a factor of 2-2.5) and decrease of general toxicity of zinc oxide compositions of doxorubicin in the form of the above mentioned gels compared to free doxorubicin were shown on the model of inoculated Ehrlich's ascitic carcinoma. Mechanistic studies of anticancer activity revealed the cytostatic effect based on the high level of DNA biosynthesis inhibition at considerable low concentrations of zinc oxide compositions of doxorubicin. The results of studies in vitro and in vivo behavior of PEO+Dox+ZnO and Starch+NaCMC+Dox+ZnO composites confirm the high potential of the nanosize zinc oxide composites as a vector delivery system for future application in cancer chemotherapy.

Keywords: anticancer activity, cancer specificity, doxorubicin, zinc oxide

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Authors: Christian Arcos, Marley Vellasco, Abraham Alcaim

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In this paper, we present a wavelet coefficients masking based on Local Binary Patterns (WLBP) approach to enhance the temporal spectra of the wavelet coefficients for speech enhancement. This technique exploits the wavelet denoising scheme, which splits the degraded speech into pyramidal subband components and extracts frequency information without losing temporal information. Speech enhancement in each high-frequency subband is performed by binary labels through the local binary pattern masking that encodes the ratio between the original value of each coefficient and the values of the neighbour coefficients. This approach enhances the high-frequency spectra of the wavelet transform instead of eliminating them through a threshold. A comparative analysis is carried out with conventional speech enhancement algorithms, demonstrating that the proposed technique achieves significant improvements in terms of PESQ, an international recommendation of objective measure for estimating subjective speech quality. Informal listening tests also show that the proposed method in an acoustic context improves the quality of speech, avoiding the annoying musical noise present in other speech enhancement techniques. Experimental results obtained with a DNN based speech recognizer in noisy environments corroborate the superiority of the proposed scheme in the robust speech recognition scenario.

Keywords: binary labels, local binary patterns, mask, wavelet coefficients, speech enhancement, speech recognition

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Authors: Janis Zarins, Kristaps Blums, Oskars Radzins, Renars Deksnis, Atis Svare, Santa Salaka

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Wide tumor resection remains the first choice method for tumors of the oral cavity. Nevertheless, remained tissue defect impacts patients functional and aesthetical outcome, which could be improved using microvascular tissue transfers. Mandibular reconstruction is challenging due to the complexity of composite tissue defects and occlusal relationships for normal eating, chewing, and pain free jaw motions. Individual 3-D virtual planning would provide better symmetry and functional outcome. The main goal of preoperative planning is to develop a customized surgical approach with patient specific cutting guides of the mandible, osteotomy guides of the fibula, pre-bended osteosynthesis plates to perform more precise reconstruction, to decrease the surgery time and reach the best outcome. Our study is based on the analysis of 32 patients operated on between 2019 to 2021. All patients underwent mandible reconstruction with vascularized fibula flaps. Patients characteristics, surgery profile, survival, functional outcome, and quality of life was evaluated. Preoperative planning provided a significant decrease of surgery time and the best arrangement of bone closely similar as before the surgery. In cases of bone asymmetry, deformity and malposition, a new mandible was created using 3D planning to restore the appearance of lower jaw anatomy and functionality.

Keywords: mandibular, 3D planning, cutting guides, fibula flap, reconstruction

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Authors: Omar Youssef, Aija Knuuttila, Paivi Piirilä, Virinder Sarhadi, Sakari Knuutila

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Exhaled breath condensate (EBC) is a unique sample that allows studying different genetic changes in lung carcinoma through a non-invasive way. With the aid of next generation sequencing (NGS) technology, analysis of genetic mutations has been more efficient with increased sensitivity for detection of genetic variants. In order to investigate the possibility of applying this method for cancer diagnostics, mutations in EBC DNA from lung cancer patients and healthy individuals were studied by using NGS. The key aim is to assess the feasibility of using this approach to detect clinically important mutations in EBC. EBC was collected from 20 healthy individuals and 9 lung cancer patients (four lung adenocarcinomas, four 8 squamous cell carcinoma, and one case of mesothelioma). Mutations in hotpot regions of 22 genes were studied by using Ampliseq Colon and Lung cancer panel and sequenced on Ion PGM. Results demonstrated that all nine patients showed a total of 19 cosmic mutations in APC, BRAF, EGFR, ERBB4, FBXW7, FGFR1, KRAS, MAP2K1, NRAS, PIK3CA, PTEN, RET, SMAD4, and TP53. In controls, 15 individuals showed 35 cosmic mutations in BRAF, CTNNB1, DDR2, EGFR, ERBB2, FBXW7, FGFR3, KRAS, MET, NOTCH1, NRAS, PIK3CA, PTEN, SMAD4, and TP53. Additionally, 45 novel mutations not reported previously were also seen in patients’ samples, and 106 novel mutations were seen in controls’ specimens. KRAS exon 2 mutations G12D was identified in one control specimen with mutant allele fraction of 6.8%, while KRAS G13D mutation seen in one patient sample showed mutant allele fraction of 17%. These findings illustrate that hotspot mutations are present in DNA from EBC of both cancer patients and healthy controls. As some of the cosmic mutations were seen in controls too, no firm conclusion can be drawn on the clinical importance of cosmic mutations in patients. Mutations reported in controls could represent early neoplastic changes or normal homeostatic process of apoptosis occurring in lung tissue to get rid of mutant cells. At the same time, mutations detected in patients might represent a non-invasive easily accessible way for early cancer detection. Follow up of individuals with important cancer mutations is necessary to clarify the significance of these mutations in both healthy individuals and cancer patients.

Keywords: exhaled breath condensate, lung cancer, mutations, next generation sequencing

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Authors: Hamdi Sana, Emna Bouazizi, Sami Faiz

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In recent years, there is a huge increase in the use of spatio-temporal applications where data and queries are continuously moving. As a result, the need to process real-time spatio-temporal data seems clear and real-time stream data management becomes a hot topic. Sliding window model and frequent itemset mining over dynamic data are the most important problems in the context of data mining. Thus, sliding window model for frequent itemset mining is a widely used model for data stream mining due to its emphasis on recent data and its bounded memory requirement. These methods use the traditional transaction-based sliding window model where the window size is based on a fixed number of transactions. Actually, this model supposes that all transactions have a constant rate which is not suited for real-time applications. And the use of this model in such applications endangers their performance. Based on these observations, this paper relaxes the notion of window size and proposes the use of a timestamp-based sliding window model. In our proposed frequent itemset mining algorithm, support conditions are used to differentiate frequents and infrequent patterns. Thereafter, a tree is developed to incrementally maintain the essential information. We evaluate our contribution. The preliminary results are quite promising.

Keywords: real-time spatial big data, frequent itemset, transaction-based sliding window model, timestamp-based sliding window model, weighted frequent patterns, tree, stream query

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Authors: Sara Mercedes Barroso Pinzón, Antonio Javier Sanchéz Herencia, Begoña Ferrari, Álvaro Jesús Castro

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The current need for durable implants and bone substitutes characterised by biocompatibility, bioactivity and mechanical properties, without immunological rejection, is a major challenge for scientists. Hydroxyapatite (HAp) has been considered for decades as an ideal biomaterial for bone regeneration due to its chemical and crystallographic similarity to the mineral structure bioapatites. However, the lack of trace elements in the hydroxyapatite structure gives it very low mechanical and biological properties. In this sense, the objective of the research is to address the synthesis of hydroxyapatite with Mg from phosphate rock from sedimentary deposits in the central-eastern region of Colombia, taking advantage of the release of the species contained as natural precursors of Ca, P and Mg. The minerals present were studied, fluorapatite as the mineral of interest associated with mineralogical species of magnesium carbonates and quartz. The chemical and mineralogical composition was determined by X-ray fluorescence (XRF) and X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX); as well as the evaluation of the surface physicochemical properties of zeta potential (PZC), with the aim of studying the surface behaviour of the microconstituents present in the phosphate rock and to elucidate the synergistic mechanism between the minerals and establish the optimum conditions for the wet concentration process. From the products obtained and characterised by XRD, XRF, SEM, FTIR, RAMAN, HAp-Mg biocomposite scaffolds are fabricated and the influence of Mg on the morphometric parameters, mechanical and biological properties of the designed materials is evaluated.

Keywords: phosphate rock, hydroxyapatite, magnesium, biomaterials

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Authors: Waleed Jarjoura

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In various cases of visual impairments, the individuals are referred to expert Ophthalmologists in order to establish a correct diagnosis. Children with visual-impairments confront various challenging experiences in life since early childhood throughout lifespan. In some cases, blind infants, especially due to congenital hydrocephalus, suffer from high intra-cranial pressure and, consequently, go through a ventriculo-peritoneal shunt surgery in order to limit the neurological symptoms or decrease the cognitive impairments. In this article, a detailed description of numerous crucial implications of the V/P shunt surgery, through the right posterior-inferior parieto-temporal cortex, on the observed preliminary capabilities that are pre-requisites for the acquisition of literacy skills in braille, basic Math competencies, braille printing which suggest Gerstmann syndrome in the blind. In addition, significant difficultiesorientation and mobility skills using the Cane, in general, organizational skills, and social interactions were observed. The primary conclusion of this report focuses on raising awareness among neuro-surgeons towards the need for alternative intracranial routes for V/P shunt implantation in blind infants that preserve the right posterior-inferior parieto-temporal cortex that is hypothesized to modulate the tactual-spatial cues in braille discrimination. A second conclusion targets educators and therapists that address the acquired dysfunctionsin blind individuals due to V/P shunt surgeries.

Keywords: congenital blindness, hydrocephalus, shunt surgery, spatial orientation

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Authors: Nino Gogatishvili, Davit Kvernadze, Giorgi Japharidze

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Background and aims: Landau Kleffner syndrome (LKS) is a rare disorder with epileptic seizures and acquired aphasia. It usually starts in initially healthy children. The first symptoms are language regression and behavioral disturbances, and the sleep EEG reveals abnormal epileptiform activity. The aim was to discuss the efficacy of Clobazam for Landau Kleffner syndrome. Case report: We report a case of an 11-year-old boy with an uneventful pregnancy and delivery. He began to walk at 11 months and speak with simple phrases at the age of 2,5 years. At the age of 18 months, he had febrile convulsions; at the age of 5 years, the parents noticed language regression, stuttering, and serious behavioral dysfunction, including hyperactivity, temper outbursts. The epileptic seizure was not noticed. MRI was without any abnormality. Neuropsychological testing revealed verbal auditory agnosia. Sleep EEG showed abundant left fronto-temporal spikes, reaching over 85% during non-rapid eye movement sleep (non-REM sleep). Treatment was started with Clobazam. After ten weeks, EEG was improved. Stuttering and behavior also improved. Results: Since the start of Clobazam treatment, stuttering and behavior improved. Now, he is 11 years old, without antiseizure medication. Sleep EEG shows fronto-temporal spikes on the left side, over 10-49 % of non-REM sleep, bioccipital spikes, and slow-wave discharges and spike-waves. Conclusions: This case provides further support for the efficacy of Clobazam in patients with LKS.

Keywords: Landau-Kleffner syndrome, antiseizure medication, stuttering, aphasia

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Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

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Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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Authors: Mace Bentley, Zhuojun Duan, Tobias Gerken, Dudley Bonsal, Henry Way, Endre Szakal, Mia Pham, Hunter Donaldson, Chelsea Lang, Hayden Abbott, Leah Wilcynzski

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Air pollution has the potential to modify the lifespan and intensity of thunderstorms and the properties of lightning. Using data mining and geovisualization, we investigate how background climate and weather conditions shape variability in urban air pollution and how this, in turn, shapes thunderstorms as measured by the intensity, distribution, and frequency of cloud-to-ground lightning. A spatiotemporal analysis was conducted in order to identify thunderstorms using high-resolution lightning detection network data. Over seven million lightning flashes were used to identify more than 196,000 thunderstorms that occurred between 2006 - 2020 in the Washington, DC Metropolitan Region. Each lightning flash in the dataset was grouped into thunderstorm events by means of a temporal and spatial clustering algorithm. Once the thunderstorm event database was constructed, hourly wind direction, wind speed, and atmospheric thermodynamic data were added to the initiation and dissipation times and locations for the 196,000 identified thunderstorms. Hourly aerosol and air quality data for the thunderstorm initiation times and locations were also incorporated into the dataset. Developing thunderstorm climatologies using a lightning tracking algorithm and lightning detection network data was found to be useful for visualizing the spatial and temporal distribution of urban augmented thunderstorms in the region.

Keywords: lightning, urbanization, thunderstorms, climatology

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Authors: Fidan Z. Aliyeva

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In the presented article, the formation of the microbiological regime in the Takhtakorpu water complex, as well as spatial-temporal changes in the quantitative indicators of bacterioplankton, were studied. Taktakorpu water complex was built as a continuation of the reconstruction and expansion project of the Samur-Absheron irrigation system in Shabran on the northeastern slope of our republic. It should be noted that with the implementation of the project, the water supply of up to 150 thousand ha of useful land in the northern region has been improved, and the drinking, technical, and irrigation water needs of the population of Baku, Sumgayit and also the Absheron Peninsula, and industrial and agricultural areas, joining the agricultural circulation of new soil areas, Takhtakorpu reservoir with a volume of 238.4 million m³, connected with them -Valvalachay- Takhtakorpu and Takhtakorpu-Jeyranbatan canals have been created, conditions have been created to increase the resources of the Jeyranbatan reservoir. Special attention is paid to the study of saprophytic bacteria in order to determine the development dynamics and biochemical activity of the microbiological regime in the Takhtakorpu Water Complex, which is of great strategic importance for our republic, to evaluate changes under the influence of anthropogenic factors, as well as to evaluate the properties of self-cleaning, mineralization features of organic substances of allochthon and autochthonous origin. One of the main goals of our research is to determine the main structural indicators of bacterioplankton in the upper part of Takhtakorpu water complex in the first three stations and analyzing their quantitative values in a certain time aspect.

Keywords: water, irrigation, sewage, wastewater

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Authors: Yun Chen, David Penton, Fazlul Karim, Santosh Aryal, Shahriar Wahid, Peter Taylor, Susan M. Cuddy

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Droughts have severely affected Afghanistan over the last four decades, leading to critical food shortages where two-thirds of the country’s population are in a food crisis. Long years of conflict have lowered the country’s ability to deal with hazards such as drought, which can rapidly escalate into disasters. Understanding the spatial and temporal distribution of droughts is needed to be able to respond effectively to disasters and plan for future occurrences. This study used Standardized Precipitation Evapotranspiration Index (SPEI) at monthly, seasonal, and annual temporal scales to map the spatiotemporal change dynamics of drought characteristics (distribution, frequency, duration, and severity) in Afghanistan. SPEI indices were mapped for river basins, disaggregated into 189 sub-catchments, using monthly precipitation and potential evapotranspiration derived from temperature station observations from 1980 to 2017. The results show these multi-dimensional drought characteristics vary along different years, change among sub-catchments, and differ across temporal scales. During the 38 years, the driest decade and period are the 2000s and 1999–2022, respectively. The 2000–01 water year is the driest, with the whole country experiencing ‘severe’ to ‘extreme’ drought, more than 53% (87 sub-catchments) suffering the worst drought in history, and about 58% (94 sub-catchments) having ‘very frequent’ drought (7 to 8 months) or ‘extremely frequent’ drought (9 to 10 months). The estimated seasonal duration and severity present significant variations across the study area and throughout the study period. The nation also suffered from recurring droughts with varying length and intensity in 2004, 2006, 2008, and, most recently, 2011. There is a trend towards increasing drought with longer duration and higher severity extending all over sub-catchments from southeast to north and central regions. These datasets and maps help to fill the knowledge gap on detailed sub-catchment scale meteorological drought characteristics in Afghanistan. The study findings improve our understanding of the influences of climate change on drought dynamics and can guide catchment planning for reliable adaptation to and mitigation against future droughts.

Keywords: SPEI, precipitation, evapotranspiration, climate extremes

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Authors: Naseem Fatima, A. N. Srivastava, Tasleem Raza, Vijay Kumar

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Introduction: Carcinoma gallbladder is third most common gastrointestinal lethal disease with the highest incidence and mortality rate among women in Northern India. Scientists have found several risk factors that make a person more likely to develop gallbladder cancer; among these risk factors, deregulation of miRNAs has been demonstrated to be one of the most crucial factors. The changes in the expression of specific miRNA genes result in the control of inflammation, cell cycle regulation, stress response, proliferation, differentiation, apoptosis and invasion thus mediate the process in tumorgenesis. The aim of this study was to investigate the role of MiRNA-335 and may as a molecular marker in early detection of gallbladder cancer in suspected cases. Material and Methods: A total of 20 consecutive patients with gallbladder cancer aged between 30-75 years were registered for the study. Total RNA was extracted from tissue by using the mirVANA MiRNA isolation Kit according to the manufacturer’s protocol. The MiRNA- 335 and U6 snRNA-specific cDNA were reverse-transcribed from total RNA using Taqman microRNA reverse-transcription kit according to the manufacturer’s protocol. TaqMan MiRNA probes hsa-miR-335 and Taqman Master Mix without AmpEase UNG, Individual real-time PCR assays were performed in a 20 μL reaction volume on a Real-Time PCR system (Applied Biosystems StepOnePlus™) to detect MiRNA-335 expression in tissue. Relative quantification of target MiRNA expression was evaluated using the comparative cycle threshold (CT) method. The correlation was done in between cycle threshold (CT Value) of target MiRNA in gallbladder cancer with respect to non-cancerous Cholelithiasis gallbladder. Each sample was examined in triplicate. The Newman-Keuls Multiple Comparison Test was used to determine the expression of miR-335. Results: MiRNA335 was found to be significantly downregulated in the gallbladder cancer tissue (P<0.001), when compared with non-cancerous Cholelithiasis gallbladder cases. Out of 20 cases, 75% showed reduced expression of MiRNA335, were at last stage of disease with low overall survival rate and remaining 25% were showed up-regulated expression of MiRNA335 with high survival rate. Conclusion: The present study showed that reduced expression of MiRNA335 is associated with the advancement of the disease, and its deregulation may provide important clues to understanding it as a prognostic marker and opportunities for future research.

Keywords: carcinoma gallbladder, downregulation, MiRNA-335, RT-PCR assay

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Authors: Ramazan F. Akkoc, Elif Erdem, Nalan Kaya, Gonca Ozan, D. Özlem Dabak, Enver Ozan

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Maternal tobacco smoke exposure is known to cause growth retardation in the neonatal skeletal system. Alpha lipoic acid, a natural antioxidant found in some foods, limits the activities of osteoclasts and supports the osteoblast's bone formation mechanism. In this study, it was aimed to investigate the effects of alpha lipoic acid (ALA) on the height, long bones and tail lengths of pups exposed to maternal tobacco smoke. The rats were divided into four groups: 1) control group, 2) tobacco smoke group, 3) tobacco smoke + ALA group, and 4) ALA group. Rats in the group 2 (tobacco smoke), group 3 (tobacco smoke + ALA) were exposed to tobacco smoke twice a day for one hour starting from eight weeks before mating and during pregnancy. In addition to tobacco smoke, 20 mg/kg of alpha lipoic acid was administered via oral gavage to the rats in the group 3 (tobacco smoke + ALA). Only alpha lipoic acid was administered to the rats in the group 4. On day 21 postpartum, the height and tail lengths of the pups in all groups were measured, and the length of the extremity long bones was measured after decapitation. All morphometric measurements performed in group 2 (tobacco smoke) showed a significant decrease compared to group 1 (control), while all measurements in group 3 (tobacco smoke + ALA) showed a significant increase compared to group 2 (tobacco smoke). It has been shown that ALA has a protective effect against the regression of height, long bones and tail lengths of pups exposed to maternal tobacco smoke.

Keywords: alpha lipoic acid, bone, morphometry, rat, tobacco smoke

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Authors: S. Alireza Mirghasemi, Javad Parvizi, Narges R. Gabaran, Shervin Rashidinia, Mahdi M. Bijanabadi, Dariush G. Savadkoohi

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Background: The anterior cruciate ligament is one of the most frequent ligament to be disrupted. Surgical reconstruction of the anterior cruciate ligament is a common practice to treat the disability or chronic instability of the knee. Several factors associated with success or failure of the ACL reconstruction including preoperative laxity of the knee, selection of the graft material, surgical technique, graft tension, and postoperative rehabilitation. We aimed to examine the biomechanical properties of any graft type and initial graft tensioning during ACL reconstruction using 3-dimensional computational finite element simulation. Methods: In this paper, 3-dimensional model of the knee was constructed to investigate the effect of graft tensioning on the knee joint biomechanics. Four different grafts were compared: 1) Bone-patellar tendon-bone graft (BPTB) 2) Hamstring tendon 3) BPTB and a band of gracilis4) Hamstring and a band of gracilis. The initial graft tension was set as “0, 20, 40, or 60N”. The anterior loading was set to 134 N. Findings: The resulting stress pattern and deflection in any of these models were compared to that of the intact knee. The obtained results showed that the combination of a band of gracilis with the former graft (BPTB or Hamstring) increases the structural stiffness of the knee. Conclusion: Required pretension during surgery decreases significantly by adding a band of gracilis to the proper graft.

Keywords: ACL reconstruction, deflection, finite element simulation, stress pattern

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Authors: Walaa Mohammed Saeed

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Studies suggest a crosstalk between bone and metabolism through Osteocalcin (OC), a bone-derived protein that plays an important role in regulating glucose and fat metabolism. Studies relate type II Diabetes Mellitus (DMII) with Homocysteine (Hcy) and cardiovascular diseases (CVD). This study investigates the relationship between levels of OC, Hcy, and DMII in 85 subjects of which 50 were diabetic female patients (29–65 years) and 35 healthy controls. OC and Hcy levels were measured in fasting blood samples using immunoassay analyzer. Fasting serum glucose, glycated hemoglobin, lipid profile, were estimated by automated Siemens Dimension XP auto-analyzer. A significant increase in the frequency of low OC levels (p < 0.001) and high Hcy levels (p < 0.001) was detected in diabetic patients compared to controls (chi-squared test). Using ANOVA test, patients were divided into tertiles based on plasma OC and Hcy levels; fasting serum glucose varied inversely with OC but directly with Hcy tertiles (p=0.049, p=0.033 respectively). Atherogenic Index of Plasma (AIP=Log TG/HDL) predicts that diabetic patients with 36% high and 15% intermediate cardiovascular risk had increased frequency of low OC levels compared to low-risk patients (p=0.047). Another group of diabetic patients with 39% high and 11% intermediate CVD risk had increased frequency of high Hcy levels (p=0.033). A significant negative correlation existed between OC and glucose (r = -0.318; p = 0.035) while correlation between glucose level and Hcy (r = 0.851 p=0.022) was positive. Hence, low serum OC levels and high Hcy levels were associated with impaired glucose metabolism that may increase cardiovascular risk in DMII.

Keywords: osteocalcin, homocysteine, type 2 diabetes, cardiovascular

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

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

Keywords: composites, gelatin, osteochondral defect, small molecule

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