Search results for: mice bone

Authors: Susana Gonçalves, Joana Lencart, Anabela Gregório Dias

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Introduction: In combination with chemotherapy, Total Body Irradiation (TBI) is most used as part of the conditioning regimen prior to allogeneic hematopoietic stem cell transplantation. Conventional TBI techniques have a long application time but non-conformality of beam-application with the inability to individually spare organs at risk. Our institution’s intention is to start using Volumetric Modulated Arc Therapy (VMAT) techniques to increase homogeneity of delivered radiation. As a first approach, a dosimetric plan was performed on a computed tomography (CT) scan of a Rando Alderson antropomorfic phantom (head and torso), using a set of six arcs distributed along the phantom. However, a full body anthropomorphic phantom is essential to carry out technique validation and implementation. Our aim is to define the physical and chemical characteristics and the ideal manufacturing procedure of upper and lower limbs to our anthropomorphic phantom, for later validate TBI using VMAT. Materials and Methods: To study the better fit between our phantom and limbs, a CT scan of Rando Alderson anthropomorphic phantom was acquired. CT was performed on GE Healthcare equipment (model Optima CT580 W), with slice thickness of 2.5 mm. This CT was also used to access the electronic density of soft tissue and bone through Hounsfield units (HU) analysis. Results: CT images were analyzed and measures were made for the ideal upper and lower limbs. Upper limbs should be build under the following measures: 43cm length and 7cm diameter (next to the shoulder section). Lower limbs should be build under the following measures: 79cm length and 16.5cm diameter (next to the thigh section). As expected, soft tissue and bone have very different electronic density. This is important to choose and analyze different materials to better represent soft tissue and bone characteristics. The approximate HU values of the soft tissue and for bone shall be 35HU and 250HU, respectively. Conclusion: At the moment, several compounds are being developed based on different types of resins and additives in order to be able to control and mimic the various constituent densities of the tissues. Concurrently, several manufacturing techniques are being explored to make it possible to produce the upper and lower limbs in a simple and non-expensive way, in order to finally carry out a systematic and appropriate study of the total body irradiation. This preliminary study was a good starting point to demonstrate the feasibility of TBI with VMAT.

Keywords: TBI, VMAT, anthropomorphic phantom, tissue equivalent materials

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Authors: Karthika Durairaj, Buvaneswari G., Gowdham M., Gilles M., Velmurugan G.

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Background: Hyperglycaemia is the primary cause of metabolic illness. Recently, researchers focused on the possibility that chemical exposure could promote metabolic disease. Hyperglycaemia causes a variety of metabolic diseases dependent on its etiologic conditions. According to animal and population-based research, individual chemical exposure causes health problems through alteration of endocrine function with the influence of microbial influence. We were intrigued by the function of gut microbiota variation in high fat and chemically induced hyperglycaemia. Methodology: C57/Bl6 mice were subjected to two different treatments to generate the etiologic-based diabetes model: I – a high-fat diet with a 45 kcal diet, and II - endocrine disrupting chemicals (EDCs) cocktail. The mice were monitored periodically for changes in body weight and fasting glucose. After 120 days of the experiment, blood anthropometry, faecal metagenomics and metabolomics were performed and analyzed through statistical analysis using one-way ANOVA and student’s t-test. Results: After 120 days of exposure, we found hyperglycaemic changes in both experimental models. The treatment groups also differed in terms of plasma lipid levels, creatinine, and hepatic markers. To determine the influence on glucose metabolism, microbial profiling and metabolite levels were significantly different between groups. The gene expression studies associated with glucose metabolism vary between hosts and their treatments. Conclusion: This research will result in the identification of biomarkers and molecular targets for better diabetes control and treatment.

Keywords: hyperglycaemia, endocrine-disrupting chemicals, gut microbiota, host metabolism

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Authors: Matej Patzelt, Jana Mrzilkova, Jan Dudak, Frantisek Krejci, Jan Zemlicka, Zdenek Wurst, Petr Zach, Vladimir Musil

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Introduction: Micro-CT is well used for examination of bone structures and teeth. On the other hand visualization of the soft tissues is still limited. The goal of our study was to elaborate methodology for soft tissue samples imaging in micro-CT. Methodology: We used organs of rats and mice. We either did a preparation of the organs and fixation in contrast solution or we did cannulation of blood vessels and their injection for imaging of the vascular system. First, we scanned native specimens, then we created corrosive specimens by resins. In the next step, we injected vascular system either by Aurovist contrast agent or by Exitron. In the next step, we focused on soft tissues contrast increase. We scanned samples fixated in Lugol solution, samples fixated in pure ethanol and in formaldehyde solution. All used methods were afterwards compared. Results: Native specimens did not provide sufficient contrast of the tissues in any of organs. Corrosive samples of the blood stream provided great contrast and details; on the other hand, it was necessary to destroy the organ. Further examined possibility was injection of the AuroVist contrast that leads to the great bloodstream contrast. Injection of Exitron contrast agent comparing to Aurovist did not provide such a great contrast. The soft tissues (kidney, heart, lungs, brain, and liver) were best visualized after fixation in ethanol. This type of fixation showed best results in all studied tissues. Lugol solution had great results in muscle tissue. Fixation by formaldehyde solution showed similar quality of contrast in the tissues like ethanol. Conclusion: Before imaging, we need to, first, determinate which structures of the soft tissues we want to visualize. In the case of the bloodstream, the best was AuroVist and corrosive specimens. Muscle tissue is best visualized by Lugol solution. In the case of the organs containing cavities, like kidneys or brain, the best way was ethanol fixation.

Keywords: experimental imaging, fixation, micro-CT, soft tissues

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Authors: Sitti Nurani Sirajuddin, Muh. Aminawar, Siti Nurlaelah, Amidah Amrawaty

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This study aimed to determine the income of beef cattle breeders from the traditional profit-sharing system named Tesang. This study was conducted in the province of South Sulawesi start from April to July 2014, used quantitative methods and data analysis is of income. The population is all beef cattle breeders who perform for the traditional profit-sharing system (Tesang) in Barru Regency and Bone Regency, province of South Sulawesi. Samples are beef cattle breeders who breeding the cattle with the traditional profit-sharing system (Tesang) in Barru Regency and Bone Regency using breeding system and cattle enlargement system (expense) by fifty breeders. The results showed beef cattle breeder’s income from the profit-sharing system (Tesang) where enlargement system (expense) at6th month maintenance periods higher than the profit-sharing system (Tesang) with using breeding the cattle.

Keywords: income, beef cattle, profit-sharing system, Teseng

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Authors: Anupuma Raina, Ajay P. Balayan, Prateek Pandya, Pankaj Shrivastava, Uma Kanga, Tulika Seth

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DNA fingerprinting analysis aids in personal identification for forensic purposes and has always been a driving motivation for law enforcement agencies in almost all countries since its inception. The introduction of DNA markers (Y-STR) has allowed for greater precision and higher discriminatory power in forensic testing. A criminal/ person committing crime after bone marrow transplantation is a rare situation but not an impossible one. Keeping such a situation in mind, a study was carried out to find out the best biological sample to be used for personal identification, especially in forensic situation. We choose a female patient (recipient) and a male donor. The pre transplant sample (blood) and post transplant samples (blood, buccal swab, hair roots) were collected from the recipient (patient). The same were compared with the blood sample of the donor using DNA FP technique. Post transplant samples were collected at different interval of time (15, 30, 60, and 90 days). The study was carried out using Y-STR kit at 23 loci. The results determined discusses the phenomenon of chimerism and its impact on Y-STR. Hair sample was found the most suitable sample which had no donor DNA profiling up to 90 days.

Keywords: bone marrow transplantation, chimerism, DNA profiling, Y-STR

Procedia PDF Downloads 133

Authors: Samar Omar A. Rabah, Sahar Ragab El Hadad, Fatmah Albani

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Since the discovery of Agarwood (Incense tree), many studies reported its characteristic effects and variable benefits, as either to produce Arabian Incense or as a traditional medicine against many diseases. Laboratory experiments were carried out on the effect of different concentrations of Incense smoke inhalation on the lung weight and tissue in female mice. This research derives its importance from the fact that Incense is heavily used in Saudi Arabia in the absence of thorough studies of its effects on health. Eighty animals are used in this study, and they are divided into four groups, each is 20 animals. Three groups are exposed to different concentrations (2, 4 and 6 gm) of Incense smoke daily for three months, and the fourth group is the control. At the end of each month, five animals from each group were dissected. Obtained data showed an increase but not significant in animal body and lung weight, this results return to natural increase as a result of normal growth of animals. Light microscope reveals some changes in the lung tissue, such as focal emphysema, rupture in the alveolar walls, hemorrhage, congestion, edema and few peri-bronchial lymphoid cells. After continuous exposure to Incense smoke focal necrosis and degradation are observed in some cells of epithelial bronchioles. Also, fibrosis of peri-bronchial, thickening in alveolar walls and aggregation of lymphoid cells are demonstrated in some lungs sections. according to the above manifestations it could be concluded that exposure to Incense smoke causes pulmonary harmful effects. Therefore, we can recommend that Incense smoke will be used only in open places to reduce its harms.

Keywords: incense smoke, lungs, histological changes of lungs, agarwood

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Authors: Mehdi Abbasi, Shadan Navid, Mohammad Pourahmadi, M. Majidi Zolbin

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We have recently reported that melatonin as antioxidant enhances the efficacy of colonization of spermatogonial stem cells (SSCs). Melatonin as an antioxidant plays a vital role in the development of SSCs in vitro. This study aimed to investigate evaluation of sertoli cells and melatonin simultaneously on SSC proliferation following transplantation to testis of adult mouse busulfan-treated azoospermia model. SSCs and sertoli cells were isolated from the testes of three to six-day old male mice.To determine the purity, Flow cytometry technique using PLZF antibody were evaluated. Isolated testicular cells were cultured in αMEM medium in the absence (control group) or presence (experimental group) of sertoli cells and melatonin extract for 2 weeks. We then transplanted SSCs by injection into the azoospermia mice model. Higher viability, proliferation, and Id4, Plzf, expression were observed in the presence of simultaneous sertoli cells and melatonin in vitro. Moreover, immunocytochemistry results showed higher Oct4 expression in this group. Eight weeks after transplantation, injected cells were localized at the base of seminiferous tubules in the recipient testes. The number of spermatogonia and the weight of testis were higher in the experimental group relative to control group. The results of our study suggest that this new protocol can increase the transplantation of these cells can be useful in the treatment of male infertility.

Keywords: colonization, melatonin, spermatogonial stem cell, transplantation

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Authors: Francisco J. Cedano, Laura M. Pinzón, Camila I. Castro, Felipe Salcedo, Juan P. Casas, Juan C. Briceño

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Complex fractures located in articular surfaces are challenging to treat and their reduction with conventional treatments could compromise the functionality of the affected limb. An adhesive material to treat those fractures is desirable for orthopedic surgeons. This adhesive must be biocompatible and have a high adhesion to bone surface in an aqueous environment. The proposed adhesive is based on chitosan, given its adhesive and biocompatibility properties. Chitosan is mixed with calcium carbonate and hydroxyapatite, which contribute to structural support and a gel like behavior, and glutaraldehyde is used as a cross-linking agent to keep the adhesive mechanical performance in aqueous environment. This work aims to evaluate the rheological, adhesion strength and biocompatibility properties of the proposed adhesive using in vitro tests. The gelification process of the adhesive was monitored by oscillatory rheometry in an ARG-2 TA Instruments rheometer, using a parallel plate geometry of 22 mm and a gap of 1 mm. Time sweep experiments were conducted at 1 Hz frequency, 1% strain and 37°C from 0 to 2400 s. Adhesion strength is measured using a butt joint test with bovine cancellous bone fragments as substrates. The test is conducted at 5 min, 20min and 24 hours after curing the adhesive under water at 37°C. Biocompatibility is evaluated by a cytotoxicity test in a fibroblast cell culture using MTT assay and SEM. Rheological results concluded that the average gelification time of the adhesive is 820±107 s, also it reaches storage modulus magnitudes up to 106 Pa; The adhesive show solid-like behavior. Butt joint test showed 28.6 ± 9.2 kPa of tensile bond strength for the adhesive cured for 24 hours. Also there was no significant difference in adhesion strength between 20 minutes and 24 hours. MTT showed 70 ± 23 % of active cells at sixth day of culture, this percentage is estimated respect to a positive control (only cells with culture medium and bovine serum). High vacuum SEM observation permitted to localize and study the morphology of fibroblasts presented in the adhesive. All captured fibroblasts presented in SEM typical flatted structure with filopodia growth attached to adhesive surface. This project reports an adhesive based on chitosan that is biocompatible due to high active cells presented in MTT test and these results were correlated using SEM. Also, it has adhesion properties in conditions that model the clinical application, and the adhesion strength do not decrease between 5 minutes and 24 hours.

Keywords: bioadhesive, bone adhesive, calcium carbonate, chitosan, hydroxyapatite, glutaraldehyde

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Authors: Shri Dubey, Jamal Ghorieshi

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Instability of the lumbar spine is caused by various factors that include degenerative disc, herniated disc, traumatic injuries, and other disorders. Pedicle screws are widely used as a main fixation device to construct rigid linkages of vertebrae to provide a fully functional and stable spine. Various technologies and methods have been used to restore the stabilization. However, loosening of pedicle screws is the main cause of concerns for neurosurgeons. This could happen due to poor bone quality with osteoporosis as well as types of pedicle screw used. Compatibilities and stabilities of pedicle screws with bone depend on design (thread design, length, and diameter) and material. Grip length and pullout strength affect the motion and stability of the spine when it goes through different phases such as extension, flexion, and rotation. Pullout strength of augmented pedicle screws is increased in both primary and salvage procedures by 119% (p = 0.001) and 162% (p = 0.01), respectively. Self-centering pedicle screws at different trajectories (0°, 10°, 20°, and 30°) show the same pullout strength as insertion in a straight-ahead trajectory. The outer cylindrical and inner conical shape of pedicle screws show the highest pullout strength in Grades 5 and 15 foams (synthetic bone). An outer cylindrical and inner conical shape with a V-shape thread exhibit the highest pullout strength in all foam grades. The maximum observed pullout strength is at axial pullout configuration at 0°. For Grade 15 (240 kg/m³) foam, there is a decline in pull out strength. The largest decrease in pullout strength is reported for Grade 10 (160 kg/m³) foam. The maximum pullout strength of 2176 N (0.32-g/cm³ Sawbones) on all densities. Type 1 Pedicle screw shows the best fixation due to smaller conical core diameter and smaller thread pitch (Screw 2 with 2 mm; Screws 1 and 3 with 3 mm).

Keywords: polymethylmethacrylate, PMMA, classical pedicle screws, CPS, expandable poly-ether-ether-ketone shell, EPEEKS, includes translaminar facet screw, TLFS, poly-ether-ether-ketone, PEEK, transfacetopedicular screw, TFPS

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Authors: Aleksandra BužArovska, Gordana Bogoeva Gaceva

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Biodegradable polymers have received significant scientific attention in tissue engineering (TE) application, in particular their composites consisting of inorganic nanoparticles. In the last 15 years, they are subject of intensive research by many groups, aiming to develop polymer scaffolds with defined biodegradability, porosity and adequate mechanical stability. The most important characteristic making these materials attractive for TE is their biodegradability, a process that could be time controlled and long enough to enable generation of a new tissue as a replacement for the degraded polymer scaffold. In this work poly(L-lactic acid) scaffolds, filled with TiO2 nanoparticles functionalized with oleic acid, have been prepared by thermally induced phase separation method (TIPS). The functionalization of TiO2 nanoparticles with oleic acid was performed in order to improve the nanoparticles dispersibility within the polymer matrix and at the same time to inhibit the cytotoxicity of the nanofiller. The oleic acid was chosen as amphiphilic molecule belonging to the fatty acid family because of its non-toxicity and possibility for mediation between the hydrophilic TiO2 nanoparticles and hydrophobic PLA matrix. The produced scaffolds were characterized with thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and mechanical compression measurements. The bioactivity for bone tissue engineering application was tested in supersaturated simulated body fluid. The degradation process was followed by Fourier transform infrared spectroscopy (FTIR). The results showed anisotropic morphology with elongated open pores (100 µm), high porosity (around 92%) and perfectly dispersed nanofiller. The compression moduli up to 10 MPa were identified independent on the nanofiller content. Functionalized TiO2 nanoparticles induced formation of hydroxyapatite clusters as much as unfunctionalized TiO2. The prepared scaffolds showed properties ideal for scaffold vascularization, cell attachment, growth and proliferation.

Keywords: biodegradation, bone tissue engineering, mineralization, PLA scaffolds

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Authors: Nonita Sarin, K.J.Singh, Anuj Kumar, Davinder Singh

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Polymeric 3D hydrogels have pivotal role in bone tissue regeneration applications. Hydrogels behave similar to the living tissues because they have large water imbibing capacity in swollen state and adjust their shape according to the tissues during tissue formation after implantation. On the other hand, hydrogels are very soft, fragile and lack mechanical strength. Incorporation of bioceramics can improve mechanical strength. Furthermore, bioceramics synthesized by sol gel technique may enhance the apatite formation and degradation rates which can lead to the increase in faster rates for new bone and tissue regeneration. Simulated body fluid (SBF) induces the poly-condensation of silanol groups which leads to formation of silica matrix and provide active sites for the precipitation of Ca2+ and PO43- ions to form apatite layer which is similar to mineral form of bone. Therefore, authors have synthesized bioceramic incorporated Polyacrylamide-carboxymethylcellulose hydrogels by free radical polymerization and bioceramic compositions of xSrO-(36-x)CaO-45SiO2-ySeO3-(12-y)P2O5-7MgO (where x=0,4 and y=0,2 mol%) were synthesized by sol gel technique. Bioceramics incorporated in polymer matrix induces quicker apatite formation during immersion in SBF by raising the pH with the release of alkaline ions during ion exchange process and the apatite formation takes place in alkaline medium. The behavior of samples PABC-0 (without bioceramics) and PABC-20 (with 20 wt% bioceramics) were evaluated by X-Ray Diffraction and FTIR. In term of bioactivity, it was observed that PABC-20 has shown hydroxyapatite (HA) formation on 1st day of immersion whereas, PABC-0 was shown apatite formation on 7th day of immersion in SBF. The rapid rate of HA growth on 1st day of immersion in SBF signifies easy regeneration of damaged bone tissues. Degradation studies have been undertaken in Phosphate Buffer Saline and PABC-20 exhibited slower degradation rate up to 9%as compared to PABC-0 up to 18%. Slower degradation rate is suitable for new tissue regeneration and cell attachment. Also, Zeta potential studies have been employed to check the surface charge and it has been observed that samples carry negative charge when immersed in SBF. In addition, the swelling test of the samples have been performed and relative swelling ratio % observed for PABC-0 is 607% and PABC-20 is 305%. This indicates that the incorporation of bioceramics leads to the filling up of the voids in between the polymer matrix which in result reduces porosity and increase the mechanical strength by filling the voids. The porosity of PABC-0 is 84% and PABC-20 is 72%. PABC-20 sample demonstrates that bioceramics incorporation reduce the porosity and improves mechanical strength. Also, maximum in vitro cell viability up to 98% with MG63 cell line has been observed which indicate that the bioceramic incorporated hydrogel(PABC-20) provide the alkaline medium which is suitable environment for cell growth.

Keywords: hydrogels, hydroxyapatite, MG63 cell line, zeta potential

Procedia PDF Downloads 131

Authors: Chengzhu Liao, Jianbo Zhang, Haiou Wang, Jing Ming, Huili Li, Yanyan Li, Hua Cheng, Sie Chin Tjong

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Since Bonfield’s group’s pioneer work, there has been growing interest amongst the materials scientists, biomedical engineers and surgeons in the use of novel biomaterials for the treatment of bone defects and injuries. This study focuses on the fabrication, mechanical characterization and biocompatibility evaluation of high density polyethylene (HDPE) reinforced with hydroxyapatite nanorods (HANR) and carbon nanofibers (CNF). HANRs of 20 wt% and CNFs of 0.5-2 wt% were incorporated into HDPE to form biocomposites using traditional melt-compounding and injection molding techniques. The mechanical measurements show that CNF additions greatly improve the tensile strength and Young’s modulus of HDPE and HDPE-20% nHA composites. Meanwhile, the nHA and CNF fillers were found to be effective to improve dimensional and thermal stability of HDPE. The results of osteoblast cell cultivation and dimethyl thiazolyl diphenyl thiazolyl tetrazolium (MTT) tests showed that the HDPE/ CNF-nHA nanocomposites are biocompatible. Such HDPE/ CNF-nHA hybrids are found to be potential biomaterials for making orthopedic joint/bone replacements.

Keywords: biocompatibility, biocomposite, carbon nanofiber, high density polyethylene, hydroxyapatite

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Authors: Carmiña L. Vargas-Zapata, María A. Conde-Sarmiento, Maria Consuelo Maestre-Vargas

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Calcium is an essential element for good growth and development of the organism, and its requirement is increased at school age. Low socio-economic populations of developing countries such as Colombia may have food deficiency of this mineral in schoolchildren that could be reflected in calcium biochemical indicators, bone alterations and anthropometric indicators. The objective of this investigation was to evaluate some calcium biochemical indicators in a group of schoolchildren of low socioeconomic level from Barranquilla city and to correlate with body mass index. 60 schoolchildren aged 7 to 15 years were selected from Jesus’s Heart Educational Institution in Barranquilla-Atlántico, apparently healthy, without suffering from infectious or gastrointestinal diseases, without habits of drinking alcohol or smoking another hallucinogenic substance and without taking supplementation with calcium in the last six months or another substance that compromises bone metabolism. The research was approved by the ethics committee at Universidad del Atlántico. The selected children were invited to donate a blood and urine sample in a fasting time of 12 hours, the serum was separated by centrifugation and frozen at ˗20 ℃ until analyzed and the same was done with the urine sample. On the day of the biological collections, the weight and height of the students were measured to determine the nutritional status by BMI using the WHO tables. Calcium concentrations in serum and urine (SCa, UCa), alkaline phosphatase activity total and of bone origin (SAPT, SBAP) and urinary creatinine (UCr) were determined by spectrophotometric methods using commercial kits. Osteocalcin and Cross-linked N-telopeptides of type I collagen (NTx-1) in serum were measured with an enzyme-linked inmunosorbent assay. For statistical analysis the Statgraphics software Centurium XVII was used. 63% (n = 38) and 37% (n = 22) of the participants were male and female, respectively. 78% (n = 47), 5% (n = 3) and 17% (n = 10) had a normal, malnutrition and high nutritional status, respectively. The averages of evaluated indicators levels were (mean ± SD): 9.50 ± 1.06 mg/dL for SCa; 181.3 ± 64.3 U/L for SAPT, 143.8 ± 73.9 U/L for SBAP; 9.0 ± 3.48 ng/mL for osteocalcin and 101.3 ± 12.8 ng/mL for NTx-1. UCa level was 12.8 ± 7.7 mg/dL that adjusted with creatinine ranged from 0.005 to 0.395 mg/mg. Considering serum calcium values, approximately 7% of school children were hypocalcemic, 16% hypercalcemic and 77% normocalcemic. The indicators evaluated did not correlate with the BMI. Low values ​​were observed in calcium urinary excretion and high in NTx-1, suggesting that mechanisms such as increase in renal retention of calcium and in bone remodeling may be contributing to calcium homeostasis.

Keywords: calcium, calcium biochemical, indicators, school children, low socioeconomic status

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Authors: Ghoul Rachid Brahim, Trad Khodja Rafik

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Vertebral localization of the hydatid cyst is a severe form of bone hydatidosis, is a parasitic infection caused by the larval forms of the tapeworms Echinococcus granulosus, The disease is slowly remaining silent (a long incubation period) which may explain why this pathology is often discovered at the stage of neurological complications. The objective of this study is to recall the clinical and radiological aspects of this condition and the importance of early diagnosis and appropriate management. We report a study of 5 patients with vertebral hydatidosis, four men and one woman, four (04) patients operated in the emergency setting for spinal cord compression (decompression by wide laminectomy with evacuation of intra and extra canal vesicles).Albendazole-based medical treatment is instituted in all patients. Results: The evolution was favorable for three patients, the other two patients reoperated for a local recurrence. Conclusion: Vertebral hydatidosis is a rare condition with a poor prognosis due to the risk of neurological damage, the infiltrating nature of bone lesions, the frequency of relapses and therapeutic difficulties. The only curative method remains surgery, which must aim for complete and large excision of the lesions as if it were a “malignant tumour”.

Keywords: hydatidosis, Echinococcosis granulosus, hydatid cyst, spinal cord compression, laminectomy

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Authors: Anuranjani Kumar, Madhu Bala

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Ionising radiation exposure induces generation of free radicals and the oxidative DNA damage. SBL-1, a radioprotective leaf extract prepared from leaves Hippophae rhamnoides L. (Common name; Seabuckthorn), showed > 90% survival in mice population that was treated with lethal dose (10 Gy) of ⁶⁰Co gamma irradiation. In this study, early effects of pre-treatment with or without SBL-1 in blood peripheral blood lymphocytes (PBMCs) were investigated by cell viability assays (trypan blue and MTT). The quantitative in vitro study of Hoescht/PI staining was performed to check the apoptosis/necrosis in PBMCs irradiated at 2 Gy with or without pretreatment of SBL-1 (at different concentrations) up to 24 and 48h. Comet assay was performed in vivo, to detect the DNA strands breaks and its repair mechanism on peripheral blood lymphocytes at lethal dose (10 Gy). For this study, male mice (wt. 28 ± 2g) were administered radioprotective dose (30mg/kg body weight) of SBL-1, 30 min prior to irradiation. Animals were sacrificed at 24h and 48h. Blood was drawn through cardiac puncture, and blood lymphocytes were separated using histopaque column. Both neutral and alkaline comet assay were performed using standardized technique. In irradiated animals, alkaline comet assay revealed single strand breaks (SSBs) that showed significant (p < 0.05) increase in percent DNA in tail and Olive tail moment (OTM) at 24 h while at 48h the percent DNA in tail further increased significantly (p < 0.02). The double strands breaks (DSBs) increased significantly (p < 0.01) at 48 h in neutral assay, in comparison to untreated control. The animals pre-treated with SBL-1 before irradiation showed significantly (p < 0.05) less DSBs at 48 h treatment in comparison to irradiated group of animals. The SBL-1 alone treated group itself showed no toxicity. The antioxidant potential of SBL-1 were also investigated by in vitro biochemical assays such as DPPH (p < 0.05), ABTS, reducing ability (p < 0.09), hydroxyl radical scavenging (p < 0.05), ferric reducing antioxidant power (FRAP), superoxide radical scavenging activity (p < 0.05), hydrogen peroxide scavenging activity (p < 0.05) etc. SBL-1 showed strong free radical scavenging power that plays important role in the studies of radiation-induced injuries. The SBL-1 treated PBMCs showed significant (p < 0.02) viability in trypan blue assay at 24-hour incubation.

Keywords: radiation, SBL-1, SSBs, DSBs, FRAP, PBMCs

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Authors: Madushani Rodrigo, Banuka Athuraliya

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In today's world of medical diagnosis and prediction, machine learning stands out as a strong tool, transforming old ways of caring for health. This study analyzes the use of machine learning in the specialized domain of sports medicine, with a focus on the timely and accurate detection of bone fractures in cricket athletes. Failure to identify bone fractures in real time can result in malunion or non-union conditions. To ensure proper treatment and enhance the bone healing process, accurately identifying fracture locations and types is necessary. When interpreting X-ray images, it relies on the expertise and experience of medical professionals in the identification process. Sometimes, radiographic images are of low quality, leading to potential issues. Therefore, it is necessary to have a proper approach to accurately localize and classify fractures in real time. The research has revealed that the optimal approach needs to address the stated problem and employ appropriate radiographic image processing techniques and object detection algorithms. These algorithms should effectively localize and accurately classify all types of fractures with high precision and in a timely manner. In order to overcome the challenges of misidentifying fractures, a distinct model for fracture localization and classification has been implemented. The research also incorporates radiographic image enhancement and preprocessing techniques to overcome the limitations posed by low-quality images. A classification ensemble model has been implemented using ResNet18 and VGG16. In parallel, a fracture segmentation model has been implemented using the enhanced U-Net architecture. Combining the results of these two implemented models, the FracXpert system can accurately localize exact fracture locations along with fracture types from the available 12 different types of fracture patterns, which include avulsion, comminuted, compressed, dislocation, greenstick, hairline, impacted, intraarticular, longitudinal, oblique, pathological, and spiral. This system will generate a confidence score level indicating the degree of confidence in the predicted result. Using ResNet18 and VGG16 architectures, the implemented fracture segmentation model, based on the U-Net architecture, achieved a high accuracy level of 99.94%, demonstrating its precision in identifying fracture locations. Simultaneously, the classification ensemble model achieved an accuracy of 81.0%, showcasing its ability to categorize various fracture patterns, which is instrumental in the fracture treatment process. In conclusion, FracXpert has become a promising ML application in sports medicine, demonstrating its potential to revolutionize fracture detection processes. By leveraging the power of ML algorithms, this study contributes to the advancement of diagnostic capabilities in cricket athlete healthcare, ensuring timely and accurate identification of bone fractures for the best treatment outcomes.

Keywords: multiclass classification, object detection, ResNet18, U-Net, VGG16

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Authors: Jie Gao

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The combination of chemotherapy with gene therapy is highly effective in cancer therapy. To achieve combined therapeutic effects in human gastric cancer over expressing EGFR, we developed targeted LPD (liposome-polycation-DNA complex) conjugated with anti-EGFR (epidermal growth factor receptor) Fab’ for co-delivery of doxorubicin (DOX) and ribonucleotide reductase M2 (RRM2) siRNA (DOX-RRM2-TLPD). The results showed that EGFR was over expressed in several gastric cancer cell lines and gastric cancer tissues. Gene Expression Omnibus (GEO) results showed that RRM2 expression was significantly higher in gastric cancer than in non-gastric cancer tissue, and RRM2 siRNA inhibited the proliferation of several gastric cancer cells, indicating that RRM2 is a candidate target for gastric cancer therapy. Confocal studies and flow cytometry showed that DOX-RRM2-TLPD delivered DOX and RRM2 siRNA to EGFR over expressing gastric cancer cells specifically and efficiently both in vitro and in vivo, resulting in enhanced therapeutic effects (cytotoxicity and apoptosis) compared with single-drug loaded or non-targeted controls, including DOX-NC-TLPD (targeted LPD co-delivering DOX and negative control siRNA), RRM2-TLPD (targeted LPD delivering RRM2 siRNA) and DOX-RRM2-NTLPD (non-targeted LPD co-delivering DOX and RRM2 siRNA). The in vivo antitumor assay showed that the average weight of the gastric cancer in mice treated with DOX-RRM2-TLPD was significantly lighter than that of mice treated with other controls. DOX-RRM2-TLPD represents an effective approach for combined therapy of gastric cancer over expressing EGFR.

Keywords: gene therapy, chemotherapy, immunoliposomes, gastric cancer

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Authors: G. A. Khalid, M. D. Jones, R. Prabhu, A. Mason-Jones, W. Whittington, H. Bakhtiarydavijani, P. S. Theobald

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Head injury in childhood is a common cause of death or permanent disability from injury. However, despite its frequency and significance, there is little understanding of how a child’s head responds during injurious loading. Whilst Infant Post Mortem Human Subject (PMHS) experimentation is a logical approach to understand injury biomechanics, it is the authors’ opinion that a lack of subject availability is hindering potential progress. Computer modelling adds great value when considering adult populations; however, its potential remains largely untapped for infant surrogates. The complexities of child growth and development, which result in age dependent changes in anatomy, geometry and physical response characteristics, present new challenges for computational simulation. Further geometric challenges are presented by the intricate infant cranial bones, which are separated by sutures and fontanelles and demonstrate a visible fibre orientation. This study presents an FE model of a newborn infant’s head, developed from high-resolution computer tomography scans, informed by published tissue material properties. To mimic the fibre orientation of immature cranial bone, anisotropic properties were applied to the FE cranial bone model, with elastic moduli representing the bone response both parallel and perpendicular to the fibre orientation. Biofiedility of the computational model was confirmed by global validation against published PMHS data, by replicating experimental impact tests with a series of computational simulations, in terms of head kinematic responses. Numerical results confirm that the FE head model’s mechanical response is in favourable agreement with the PMHS drop test results.

Keywords: finite element analysis, impact simulation, infant head trauma, material properties, post mortem human subjects

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Authors: Sanjay Shukla

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Excessive use of antibiotics is a main problem in the treatment of wounds and other chronic infections and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most effective approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of current study was to investigate the efficiency of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in double agar overlay method out of 150 sewage samples. In TEM recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9 and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate was very safe, did not show any appearance of abscess formation which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureus which indicates that they are good prophylactic agent. The S. aureus inoculated mice were completely recovered by bacteriophage administration with 100% recovery which was very good as compere to conventional therapy. In present study ten chronic cases of wound were treated with phage lysate and follow up of these cases was done regularly up to ten days (at 0, 5 and 10 d). Result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for treatment of septic chronic wounds.

Keywords: phage therapy, phage lysate, antimicrobial resistance, S. aureus

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Authors: S. A. Hashemvarzi, A. Heidarianpour, Z. Fallahmohammadi, M. Pourghasem, M. Kaviani

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Introduction: Parkinson’s disease (PD) is a progressive neurodegenerative in the central nervous system characterized by the loss of dopaminergic neurons in the substantia nigra resulting in loss of dopamine release in the striatum. Non-drug treatment options such as Stem cell transplantation and exercise have been considered for treatment of Parkinson's disease. Purpose: The purpose of this study was to evaluate the effect of aerobic exercise after bone marrow stem cells transplantation on recovery of dopaminergic neurons and promotion of angiogenesis markers in the striatum of parkinsonian rats. Materials and Methods: 42 male Wistar rats were divided randomly into six groups: Normal (N), Sham (S), Parkinson’s (P), Stem cells transplanted Parkinson’s (SP), Exercised Parkinson’s (EP) and Stem cells transplanted + Exercised Parkinson’s (SEP). To create a model of Parkinson's, the striatum was destroyed by injection of 6-hydroxy-dopamine into the striatum through stereotaxic apparatus. Stem cells were derived from the bone marrow of femur and tibia of male rats with 6-8 weeks old. After cultivation, approximately 5×105 cells in 5 microliter of medium were injected into the striatum of rats through the channel. Aerobic exercise was included 8 weeks of running on the treadmill with a speed of 15 meters per minute. At the end, all subjects were decapitated and striatum tissues were separately isolated for measurement of vascular endothelial growth factor (VEGF), dopamine (DA) and tyrosine hydroxylase (TH) levels. Results: VEGF, DA and TH levels in the striatum of parkinsonian rats significantly increased in treatment groups (SP, EP and SEP), especially in SEP group compared to P group after treatment (P<0.05). Conclusion: The findings implicate that the BMSCs transplantation in combination with exercise would have synergistic effects leading to functional recovery, dopaminergic neurons recovery and promotion of angiogenesis marker in the striatum of parkinsonian rats.

Keywords: stem cells, treadmill training, neurotrophic factors, Parkinson

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Authors: Hager Elsheikh, Matthias Sendler, Juliana Glaubnitz

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In pancreatitis, an inflammatory reaction occurs in the pancreatic secretory cells due to premature activation of proteases, leading to pancreatic self-digestion and necrotic cell death of acinar cells. Acute pancreatitis in patients is characterized by a severe immune reaction that could lead to serious complications, such as organ failure or septic shock, if left untreated. Chronic pancreatitis is a recurrence of episodes of acute pancreatitis resulting in a fibro-inflammatory immune response, in which the type 2 immune response is primarily driven by AAMs in the pancreas. One of the most important signaling pathways for M2 macrophage activation is the IL-4/STAT6 pathway. Pancreatic fibrosis is induced by the hyperactivation of pancreatic stellate cells by dysregulation in the inflammatory response, leading to further damage, autodigestion and possibly necrosis of pancreatic acinar cells. The aim of this research is to investigate the effect of STAT6 knockout in disease severity and development of fibrosis wound healing in the presence of different macrophage populations, regulated by the type 2 immune response, after inducing chronic and/or acute pancreatitis in mice models via cerulean injection. We further investigate the influence of the JAK/STAT6 signaling pathway on the balance of fibrosis and regeneration in STAT6 deficient and wild-type mice. The characterization of resident and recruited macrophages will provide insight into the influence of the JAK/STAT6 signaling pathway on infiltrating cells and, ultimately, tissue fibrosis and disease severity.

Keywords: acute and chronic pancreatitis, tissue regeneration, macrophage polarization, Gastroenterology

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Authors: Udoaka A. I., Didia B. C.

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Background: The maturation of a child’s bones as it grows to adulthood can be viewed radiologically. The skeletal age (bone age) is the average age at which a particular stage of bone maturation is achieved. The Greulich and Pyle standard is the commonest method used to assess the skeletal age using the hand and wrist radiograph throughout the world. This atlas was compiled solely from Caucasian children and made use of the orderly sequence of carpal ossification to determine the skeletal age. Several authors have faulted this atlas for not being suitable for other races. Aim: The aim of this study is to determine if the Greulich and Pyle Atlas is applicable to Nigerian children when compared to their chronological ages. Methods: The total number of 78 normal radiographs of the hand and wrist of Nigerian children obtained from several hospitals were used for this study . These radiographs were compared with the atlas and their skeletal ages noted form the atlas. The child’s chronological age in each case was also recorded. Results: The result shows a mean increase of two months in the skeletal ages of the Nigerian children compared to the atlas. This difference, however, was not significant. The skeletal age (in months) was greater in 77% of the children than the expected age in the atlas. Conclusion: The mean skeletal age of Nigerian children, though more than the standard in the atlas, is not statistically significant; as a result the study finds the radiographic atlas of Greulich and Pyle atlas applicable to Nigerian children.

Keywords: Greulich and Pyle Atlas, radiograph, skeletal age

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Authors: Mohamad Ahrari, Kayvan Sadri, Mahmoud Reza Jafari

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PEGylated liposomes have a smaller volume of distribution and decreased clearance, consequently, due to their more prolonged presence in bloodstream and maintaining their stability during this period, these liposomes can be applied for imaging tumoral sites. The purpose of this study is to develop an appropriate radiopharmaceutical agent in long-term imaging for improved diagnosis and evaluation of tumors. In this study, liposomal formulations encapsulating albumin is synthesized by solvent evaporation method along with homogenization, and their characteristics were assessed. Then these liposomes labeled by Philips method and the rate of stability of labeled liposomes in serum, and ultimately the rate of biodistribution and gamma scintigraphy in C26-colon carcinoma tumor-bearing mice, were studied. The result of the study of liposomal characteristics displayed that capable of accumulating in tumor sites based of EPR phenomenon. these liposomes also have high stability for maintaining encapsulated albumin in a long time. In the study of biodistribution of these liposomes in mice, they accumulated more in the kidney, liver, spleen, and tumor sites, which, even after clearing formulations in the bloodstream, they existed in high levels in these organs up to 96 hours. In gamma scintigraphy also, organs with high activity accumulation from early hours up to 96 hours were visible in the form of hot spots. concluded that PEGylated liposomal formulation encapsulating albumin can be labeled with In-Oxine, and obtained stabilized formulation for long-term imaging, that have more favorable conditions for the evaluation of tumors and it will cause early diagnosis of tumors.

Keywords: nano liposome, 111In-oxine, imaging, biodistribution, tumor

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Authors: P.J. Jiji, B.V. Murlimanju, Latha V. Prabhu, Mangala M. Pai

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Background: The lunate bone dislocation can lead to the compression of the median nerve and subsequent carpal tunnel syndrome. The dislocation can interrupt the vasculature and would cause avascular necrosis. The objective of the present study was to study the morphology and number of the nutrient foramina in the cadaveric dried lunate bones of the Indian population. Methods: The present study included 28 lunate bones (13 right sided and 15 left sided) which were obtained from the gross anatomy laboratory of our institution. The bones were macroscopically observed for the nutrient foramina and the data was collected with respect to their number. The tabulation of the data and analysis were done. Results: All of our specimens (100%) exhibited the nutrient foramina over the non-articular surfaces. The foramina were observed only over the palmar and dorsal surfaces of the lunate bones. The foramen ranged between 2 and 10. The foramina were more in number over the dorsal surface (average number 3.3) in comparison to the palmar surface (average number 2.4). Conclusion: We believe that the present study has provided important data about the nutrient foramina of the lunate bones. The data is enlightening to the orthopedic surgeon and would help in the hand surgeries. The morphological knowledge of the vasculature, their foramina of entry and their number is required to understand the concepts in the lunatomalacia and Kienbock’s disease.

Keywords: avascular necrosis, foramen, lunate, nutrient

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Authors: Chu Wan-Loy, Kok Yih-Yih, Choong Siew-Ling

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The human health risks due to poor air quality caused by a wide array of microorganisms have attracted much interest. Airborne algae have been reported as early as 19th century and they can be found in the air of tropic and warm atmospheres. Airborne algae normally originate from water surfaces, soil, trees, buildings and rock surfaces. It is estimated that at least 2880 algal cells are inhaled per day by human. However, there are relatively little data published on airborne algae and its related adverse health effects except sporadic reports of algae associated clinical allergenicity. A collection of airborne algae cultures has been established following a recent survey on the occurrence of airborne algae in indoor and outdoor environments in Kuala Lumpur. The aim of this study was to investigate the allergenic potential of the isolated airborne green and blue-green algae, namely Scenedesmus sp., Cylindrospermum sp. and Hapalosiphon sp.. The suspensions of freeze-dried airborne algae were adminstered into balb-c mice model through intra-nasal route to determine their allergenic potential. Results showed that Scenedesmus sp. (1 mg/mL) increased the systemic Ig E levels in mice by 3-8 fold compared to pre-treatment. On the other hand, Cylindrospermum sp. and Hapalosiphon sp. at similar concentration caused the Ig E to increase by 2-4 fold. The potential of airborne algae causing Ig E mediated type 1 hypersensitivity was elucidated using other immunological markers such as cytokine interleukin (IL)- 4, 5, 6 and interferon-ɣ. When we compared the amount of interleukins in mouse serum between day 0 and day 53 (day of sacrifice), Hapalosiphon sp. (1mg/mL) increased the expression of IL4 and 6 by 8 fold while the Cylindrospermum sp. (1mg/mL) increased the expression of IL4 and IFɣ by 8 and 2 fold respectively. In conclusion, repeated exposure to the three selected airborne algae may stimulate the immune response and generate Ig E in a mouse model.

Keywords: airborne algae, respiratory, allergenic, immune response, Malaysia

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Authors: Ching-Yuan Chen, Lung-Kwang Pan

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Objective: Back pain is a major health problem costing billions of health budgets annually in Taiwan. Thousands of back pain surgeries are performed annually with up to 40% of patients complaining of back pain at time of post-surgery causing failed back surgery syndrome (FBSS), although diagnosis in these patients may be complex. The aim of study is to assess the feasibility of using bone SPECT-CT imaging to localize the active lesions causing persistent, recurrent or new backache after spine surgery. Materials and Methods: Bone SPECT-CT imaging was performed after the intravenous injection of 20 mCi of 99mTc-MDP for all the patients with diagnosis of FBSS. Patients were evaluated using status of subjectively pain relief, functional improvement and degree of satisfaction by reviewing the medical records and questionnaires in a 2 more years’ follow-up. Results: We enrolled a total of 16 patients were surveyed in our hospital from Jan. 2015 to Dec. 2016. Four people on SPEC/CT imaging ensured significant lesions were undergone a revised surgery (surgical treatment group). The mean visual analogue scale (VAS) decreased 5.3 points and mean Oswestry disability index (ODI) improved 38 points in the surgical group. The remaining 12 on SPECT/CT imaging were diagnosed as no significant lesions then received drug treatment (medical treatment group). The mean VAS only decreased 2 .1 point and mean ODI improved 12.6 points in the medical treatment group. In the posttherapeutic evaluation, the pain of the surgical treatment group showed a satisfactory improvement. In the medical treatment group, 10 of the 12 were also satisfied with the symptom relief while the other 2 did not improve significantly. Conclusions: Findings on SPECT-CT imaging appears to be easily explained the patients' pain. We recommended that SPECT/CT imaging was a feasible and useful clinical tool to improve diagnostic confidence or specificity when evaluating patients with FBSS.

Keywords: failed back surgery syndrome, oswestry disability index, SPECT-CT imaging, 99mTc-MDP, visual analogue scale

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Authors: Pármenas Costa Macedo do Nascimento

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Periodontal disease is the most common disease in the oral cavity of felines. It starts with the accumulation of bacteria on the tooth surface supporting the tissues of the periodontal tissue, namely gums, alveolar bone, cementum, and periodontal ligament. The main clinical symptom observed by the owner is bad breath, which may lead to local and systemic consequences depending on the stage of periodontal disease, such as bleeding and bone loss. Therefore, the study is important to educate tutors to take better care of the felines oral health in order to try to prevent the disease. For this epidemiological study, the target population has been felines, located on the outskirts of Manaus, in the state of Amazonas, with a geographic area of 155.68 km², with no defined breed, from October 1st to 10th, 2021, whose samples has been randomly selected, with a detailed profile. The variables of interest for this study have been: absence or presence of periodontal disease, gender, age (delimited by age group), and condition (domiciled or homeless). Using a sample of 40 felines from 4 districts of the east side of Manaus chosen at random, an oral exam has been made to identify the studied disease. The animal's apparent age, condition, sex, and presence or absence of periodontal disease has been noted. It has been observed that 70% (28/40) of them had periodontal disease, mostly females, aged between 0 and 5 years and domiciled, totaling 30% (12/40).

Keywords: felines, oral cavity, oral exam, periodontal disease

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Authors: Yuncang Li, Cuie Wen

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Biodegradable metallic materials such as magnesium (Mg)-based alloys have attracted extensive interest for use as bone implant materials. However, the high biodegradation rate of existing Mg alloys in the physiological environment of human body leads to losing mechanical integrity before adequate bone healing and producing a large volume of hydrogen gas. Therefore, slowing down the biodegradation rate of Mg alloys is a critical task in developing new biodegradable Mg alloy implant materials. One of the most effective approaches to achieve this is to strategically design new Mg alloys with low biodegradation rate, excellent biocompatibility, and enhanced mechanical properties. Our research selected biocompatible and biofunctional alloying elements such as zirconium (Zr), strontium (Sr), and rare earth elements (REEs) to alloy Mg and has developed a new series of Mg-Zr-Sr-REEs alloys for biodegradable implant applications. Research results indicated that Sr and Zr additions could refine the grain size, decrease the biodegradation rate, and enhance the biological behaviors of the Mg alloys. The REE addition, such as holmium (Ho) and dysprosium (Dy) to Mg-Zr-Sr alloys resulted in enhanced mechanical strength and decreased biodegradation rate. In addition, Ho and Dy additions (≤ 5 wt.%) to Mg-Zr-Sr alloys led to enhancement of cell adhesion and proliferation of osteoblast cells on the Mg-Zr-Sr-Ho/Dy alloys.

Keywords: biocompatibility, magnesium, mechanical and biodegrade properties, rare earth elements

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Authors: Muhammad Saleem

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Background: Inattentive food choices and engagement in excessive physical activities by male athletes can potentially lead to adverse health consequences. Objective: The aim was to ascertain the occurrence of risk factors associated with the Male Athlete Triad among young elite athletes in Pakistan. Methodology: In 2018, a cross-sectional study based on questionnaires was conducted at the Pakistan Sports Board. The study aimed to explore the risk factors related to the Male Athlete Triad in young elite athletes who were part of national training camps in major metropolitan areas. The study included proficient male elite athletes aged 18 to 25 years, capable of understanding the English questionnaire. The athletes completed a survey encompassing aspects like demographic information, educational background, Body Mass Index (BMI), sports involvement, and hours of participation. Additionally, they filled out the Eating Attitude Test-26 (EAT-26) and questionnaires assessing risks of amenorrhea and low bone mineral density. The prevalence of risk factors for each of the three components was individually evaluated. The collected data underwent analysis using SPSS-20, with descriptive statistics being applied. Results: The study comprised a sample of 90 elite athletes (mean age: 23.57 ± 2.37 years, mean BMI: 21.97 ± 1.90) engaged in various sports. The EAT-26 results indicated that 50% of athletes were at risk of developing an eating disorder. Moreover, 83.3% exhibited disordered eating behaviors that necessitated referral. Risks for amenorrhea were observed in 15% of the participants, and regarding low bone mineral density, notable risks were absent except for the consumption of caffeinated beverages, which was noted in 51.7% of participants. Conclusion: The study identified a significant prevalence of disordered eating risk among male elite athletes in Pakistan. However, the risks associated with amenorrhea and low bone mineral density were not a major concern in this particular group.

Keywords: 1. health and physical education risk factors male athlete associated with the male athlete traid in young elite athlete from pakistan., 2. sports sciences pakistan, 3. risk factors sports sciences pakistan, 4. triad and young elite athlete from pakistan

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Authors: Lukas Ded

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Studying the tissue structure and histogenesis in the natural, 3D context is challenging but highly beneficial process. Contrary to classical approach of the physical tissue sectioning and subsequent imaging, it enables to study the relationships of individual cellular and histological structures in their native context. Recent developments in the tissue clearing approaches and microscopic volume imaging/data processing enable the application of these methods also in the areas of developmental and reproductive biology. Here, using the CLARITY tissue procedure and 3D confocal volume imaging we optimized the protocol for clearing, staining and imaging of the mice seminiferous tubules isolated from the testes without cardiac perfusion procedure. Our approach enables the high magnification and fine resolution axial imaging of the whole diameter of the seminiferous tubules with possible unlimited lateral length imaging. Hence, the large continuous pieces of the seminiferous tubule can be scanned and digitally reconstructed for the study of the single tubule seminiferous stages using nuclear dyes. Furthermore, the application of the antibodies and various molecular dyes can be used for molecular labeling of individual cellular and subcellular structures and resulting 3D images can highly increase our understanding of the spatiotemporal aspects of the seminiferous tubules development and sperm ultrastructure formation. Finally, our newly developed algorithms for 3D data processing enable the massive parallel processing of the large amount of individual cell and tissue fluorescent signatures and building the robust spermatogenic models under physiological and pathological conditions.

Keywords: CLARITY, spermatogenesis, testis, tissue clearing, volume imaging

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