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In recent years, there has been a growing interest in 3D printing technology within the field of bioengineering. This technology offers the ability to create devices with intricate macro- and micro-geometries, as well as specific models. It has particularly gained attention for its potential in personalized medicine, allowing for the production of organ or tissue models tailored to individual patient needs. Further, 3D printing has opened up possibilities to manufacture structures that can substitute, complement, or enhance damaged or dysfunctional organic parts. To apply 3D printing in the medical field, researchers have studied various materials known as biomaterials, each with distinct chemical and physical characteristics. These materials fall into two main categories: hard and soft materials. Each biomaterial needs to possess specific characteristics that are compatible with biological systems, ensuring long-term stability and biocompatibility. In this paper, we aim to review some of the materials used in the biomedical field, with a particular focus on those utilized in veterinary medicine and ophthalmology. We will discuss the significant findings from recent scientific research, focusing on the biocompatibility, structure, applicability, and in vitro and in vivo biological characteristics of two hard and four soft materials. Additionally, we will present the current state and prospects of veterinary ophthalmology.

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The aim of this study is to report the results and to review the outcome of 14 cases of Y-T humeral fractures repair using paired polyaxial locking system (PAX) plates through a combined medial and lateral approach. Fourteen consecutive dogs, with traumatic humeral Y-T fractures, met the inclusion criteria. This study includes signalment, preoperative radiographs, type of implants, radiographic bone healing assessment, complications, range of motion (ROM) of the elbow and limb function evaluated at 120 days after surgery. Postoperative radiographs revealed adequate anatomic reconstruction, and in all cases, bone healing has been achieved. No implant failure was observed. Functional outcome was excellent in 7 dogs (no lameness and preserved ROM), good in 4 (slight lameness and moderate ROM reduction) and discrete in 2 (mild lameness and severe ROM reduction). Complications were encountered in 2/14 patients with implant-associated infection resolved after long-term antibiotic treatment and implant removal. The PAX system is shown to be a valid alternative for the treatment of Y-T humeral fractures, offering the benefit of polyaxial insertion of locking screws. The possibility of angle locking screws is helpful in the distal humeral bicondylar fractures, providing additional options for screw placement in juxtarticular fractures, avoiding fracture lines or other implants.

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Background: Tibiotarsal instabilities caused by partial or complete rupture of the medial collateral tarsal ligament (MCTL) are commonly treated by arthrodesis techniques with poor functional results and significant complication rates. Case Description: This study describes a new surgical technique for synthetic reconstruction of the MCTL in an overweight dog (estimated body condition score 8/9) with an avulsion of the long head of the MCTL. Three bone tunnels were drilled in the distal tibia, the talus, and the central tarsal bone, thus respecting the anatomical insertions of the physiological ligament. An Ultra-High Molecular Weight Polyethylene (UHMWPE) implant was fixed with interference screws to reconstruct the long and short heads of the MCTL. Premature weight-bearing was reported at 2 weeks postoperatively following early removal of the flexible restraint (bivalve resin boot), which had initially been prescribed for 6 weeks. At 11 weeks postoperatively, the tibiotarsal joint showed good valgus stability and the dog's gait was subnormal. At 12 and 16 months postoperatively, the dog regained full function of the operated limb although no weight loss was initiated as recommended. Conclusion: The use of a UHMWPE implant fixed with interference screws to reconstruct the MCTL allowed a return to full function of the tibiotarsal joint, without complications despite an early return to weight-bearing without external restraint. The success of this isolated surgical technique could lead to improvements in the surgical management of MCTL rupture if these initial results are confirmed by a prospective study with a larger number of patients.

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The objective of this study is to evaluate the difference between the amount of bone visible with the superimposition of a radiolucent hybrid external fixator and a graphically simulated metallic frame. Eighteen frames were applied to eighteen bone specimens. The fracture area (FA), the radiolucent area (RLA) and the radiopaque area (ROA) inside the FA were calculated for each construct on both postoperative views. The ratio between the RLA and FA and between the ROA and FA was used to evaluate the amount of bone visible in the FA with a radiolucent and a radiopaque fixator, respectively. Finally, the areas of RLA and ROA were compared using the Wilcoxon test and Friedman test to evaluate the effect of the radiolucent material on the amount of bone visible. Differences were considered significant if p < 0.5. In every specimen p was <0.5. The amount of bone visible was significantly higher with the radiolucent frame compared to the radiopaque frame. Based on the results of this study, the use of radiolucent materials can be a valuable option for external fixation, in order to decrease the radiographic interference of the frame, allowing better assessment of fracture reduction and bone healing on postoperative radiographs.

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An efficient analytical strategy based on different extraction methods of biologically active naturally occurring oxyprenylated umbelliferone and ferulic acid derivatives 7-isopentenyloxycoumarin, auraptene, umbelliprenin, boropinic acid, and 4'-geranyloxyferulic acid and quantification by UHPLC with spectrophotometric (UV/Vis) detection from Tea tree oil is reported. Absorption of the pure oil on Al2O3 (Brockmann activity II) prior washing the resulting solid with MeOH and treatment of this latter with CH2Cl2 resulted the best extraction methodology in terms of yields of oxyprenylated secondary metabolites. Among the five O-prenylphenylpropanoids herein under investigation auraptene and umbelliprenin were never detected while 4'-geranyloxyferulic acid was the most abundant compound resulting from all the three extraction methods employed. The UHPLC analytical methodology set up in the present study resulted to be an effective and versatile technique for the simultaneous characterization and quantification of prenyloxyphenylpropanoids in Tea tree oil and applicable to other complex matrices from the plant kingdom.

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OBJECTIVES: The aim of the study was to describe the differences in pelvic limb alignment between healthy purebred English Bulldogs and those with medial patellar luxation through the measurement of femoral and tibial angles on computed tomography images in multiplanar reconstruction modality (MPR-CT). METHODS: Twenty-one purebred English Bulldogs were included and divided into two groups: one including healthy dogs (15 limbs) and the other including those with medial patellar luxation (24 limbs). Three different observers used MPR-CT to measure the following angles: anatomical lateral proximal femoral angle (aLPFA), anatomical lateral distal femoral angle (aLDFA), mechanical lateral proximal femoral angle (mLPFA), mechanical lateral distal femoral angle (mLDFA), angle of inclination of the femoral neck (AI), angle of anteversion (AA), mechanical medial proximal tibial angle (mMPTA), mechanical medial distal tibial angle (mMDTA), mechanical caudal distal tibial angle (mCdDTA), mechanical caudal proximal tibial angle (mCdPTA), and the tibial torsion angle (TTA). A Mann-Whitney U test was used to compare each variable in both groups. RESULTS: The values for aLDFA and mLDFA in the medial patellar luxation population were significantly increased compared to healthy subjects (p <0.05). No significant differences were observed for the other variables. CLINICAL SIGNIFICANCE: In our population, an increased distal femoral varus was associated with medial patellar luxation. Our results could be useful to determine whether or not angular deformity of the femur is present and help determine the degree of correction necessary to restore alignment.

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OBJECTIVE: Evaluation of the bending properties in one direction of three titanium polyaxial locking plate systems. MATERIALS AND METHODS: The Polyaxial Advanced Locking System (PAX®) straight plate (PAX SP), the PAX® reconstruction plate (PAX RP), and the VetLOX reconstruction plates (VetLOX) were evaluated individually and as constructs applied to a bone model simulating a fracture gap and compared using a two-way analysis of variance and Tukey post-hoc analysis. RESULTS: The PAX SP had the highest values of bending stiffness, bending structural stiffness and bending strength. When tested as plates alone, the PAX RP and VetLOX showed no differences with regard to bending stiffness and bending structural stiffness, whilst the PAX RP had significantly higher strength. The PAX RP construct had significantly higher bending stiffness, bending structural stiffness and bending strength than the VetLOX construct. CLINICAL RELEVANCE: The PAX RP and VetLOX reconstruction plates are much more likely to fail when used as bridging implants, thus adjunct support is needed. The lower bending strength of the VetLOX reconstruction plates suggests it should not be used in fractures under high loads.

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Canine cranial cruciate ligament rupture (CrCLR) is a very common pathology. Surgical stabilization is the first choice treatment, although it does not fully eliminate the increased risk of osteoarthritis. This preliminary study was carried out to explore whether a newly formulated joint health supplement would benefit metabolic, clinical and radiographic changes in dogs with CrCLR surgically treated with tibial plateau leveling osteotomy (TPLO). Besides chondroitin sulfate and glucosamine hydrochloride, the studied supplement contained anti-inflammatory and antioxidant ingredients, the main ones being N-palmitoyl-D-glucosamine (Glupamid®) and quercetin. It was thus intended to target not only chondrodegenerative components of osteoarthritis, but also post-injury inflammatory response and oxidative stress of joint tissues. Thirteen dogs underwent TPLO and were randomly allocated to treatment (n = 6) and control groups (n = 7), the former receiving the joint supplement for 90 days. Lameness and radiographic osteoarthritis changes were scored before (i.e., baseline) and at 30 and 90 days post-surgery. Synovial fluid samples were collected from injured stifles at the same time points. Levels of representative metabolites were measured by proton nuclear magnetic resonance spectroscopy in a blinded fashion. In the metabolomic analysis, special attention was paid to lactate, due to its emerging recognition as a key marker of inflammation. In the last time period (from the 30th to the 90th day), lameness improved by a factor of 2.3 compared to control dogs. No significant difference was observed in the radiographic osteoarthritis score between groups. In the first postoperative month, lactate and creatine levels significantly dropped in treated compared to control dogs. Compared to surgery alone, combining the joint supplement with TPLO resulted in a trend to a better clinical outcome in the later time interval but did not influence osteoarthritis radiographic progression. A significantly better rebalance of joint microenvironment in the early time interval (baseline - 30 days) was shown by metabolomic analysis, thus suggesting that the study supplement could limit ongoing inflammatory responses.

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OBJECTIVE: Temozolomide (TMZ) improves Glioblastoma Multiforme (GBM) patient survival. The invasive behavior of the glioma cells is the cause of GBM relapse. The glioma BK ion channel (gBK) may provide glioma cells with a mechanism to invade surrounding tissue. gBK contains epitopes that cytolytic T lymphocytes (CTLs) can recognize and kill glioma cells. Fascin-1 is an actin crosslinking molecule that supports microvilli; these membrane protrusions provide a physical defense against CTLs. TMZ was investigated to determine its effect on gBK and fascin-1 expression. RESEARCH DESIGN AND METHODS: Human glioma cells cultured in TMZ were analyzed for their altered mRNA and gBK protein levels by using quantitative real time PCR, immunostaining and cellular functional assays. RESULTS: TMZ slowed glioma cell growth and inhibited their transmigratory properties due to loss of fascin-1. TMZ induced increased gBK and HLA expression and allowed these TMZ-treated cells to become better targets for gBK-specific CTLs. CONCLUSIONS: Besides its traditional chemotherapeutic effect, TMZ can have four other targeted pathways: 1) slowed glioma cell growth; 2) inhibited glioma cell transmigration; 3) increased HLA-A2 and gBK tumor antigen production; 4) increased CTL-mediated cytolysis of the TMZ treated glioma cells due to the loss of their defensive membrane protrusions supported by fascin-1.

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Increased levels of asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA) and low plasma level of L-arginine (L-ARG) are all conditions likely to decrease nitric oxide (NO) production. Aim of this study is to evaluate ADMA, SDMA, and L-ARG plasmatic levels before and after physical exercise in patients with coronary artery disease (CAD). We studied 30 patient with mean age 52 + 4.5 years. After inclusion in the study, before the execution of physical exercise, heparinized blood sample was drawn from an indwelling arterial line for determination of ADMA, L-ARG and SDMA (baseline values). Subsequently a blood sample was drawn after the physical exercise. The mean plasma concentrations of ADMA (0.68 + 0.06 vs 0.48 + 0.05 µmol/L) and SDMA (0.45 + 0.03 vs 0.30 + 0.03 µmol/L) were significantly lower after physical exercise in comparison to baseline value, while L-ARG mean levels were increased (44.20 + 10.5 vs 74.13 + 11.2 µmol/L). Physical exercise has a beneficial effect by reducing plasmatic ADMA and SDMA levels, and increasing L-ARG substrate for endothelial NO.

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Cancer cells derived from Glioblastoma multiforme possess membranous protrusions allowing these cells to infiltrate surrounding tissue, while resisting lymphocyte cytotoxicity. Microvilli and filopodia are supported by actin filaments cross-linked by fascin. Fascin-1 was genetically silenced within human U251 glioma cells; these knock-down glioma cells lost their microvilli/filopodia. The doubling time of these fascin-1 knock-down cells was doubled that of shRNA control U251 cells. Fascin-1 knock-down cells lost their transmigratory ability responding to interleukin-6 or insulin-like growth factor-1. Fascin-1 silenced U251 cells were more easily killed by cytolytic lymphocytes. Fascin-1 knock-down provides unique opportunities to augment glioma immunotherapy by simultaneously targeting several key glioma functions: like cell transmigration, cell division and resisting immune responses.

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A 7-year-old dog was presented with a history of an open lesion on the right thoracic wall, discharging honey-like fluid and small stones. Ultrasonography and computed tomographic fistulography identified a cholecystocutaneous fistula; cholecystectomy was curative. Veterinarians should consider this disease in patients with long-term discharging lesions on the right thoracic or abdominal wall.

Fistule cholécystocutanée contenant des cholélithes multiples chez un chien. Un chien âgé de 7 ans a été présenté avec une anamnèse de lésion ouverte sur la paroi thoracique droite, d'où s'écoulait un liquide ressemblant à du miel et de petits calculs. Une échographie et une fistulographie tomographique ont identifié une fistule cholécystocutanée et une cholécystectomie a guéri l'affection. Les vétérinaires devraient considérer cette maladie chez les patients ayant des lésions suppurantes de longue date sur la paroi thoracique ou abdominale droite.(Traduit par Isabelle Vallières).

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The inactivation of the homotetrameric cytosolic alcohol dehydrogenase I from Kluyveromyces lactis (KlADH I) by naturally occurring disulfides, oxidized glutathione, cystine and cystamine, was studied. The inactivation was fully reversed by dithiothreitol. The nicotinamide coenzyme, but not the substrate ethanol, protected KlADH I from inactivation. Gel filtration experiments and SDS-PAGE analysis, also, revealed that enzyme inactivation coincides with inter-subunits disulfide bond formation which are noticeably enhanced after prolonged oxidation with GSSG. Moreover, oxidized KlADH I, as its reduced state, retained the tetrameric stucture and appears mainly as a dimer under non-reducing SDS-PAGE. Conversely, KlADH I Cys278Ile mutant is unaffected by disulfides treatment. Therefore, in vitro, KlADH I wild-type could exist in two reversible forms: reduced (active) and oxidized (inactive), in which the Cys278 residues of each tetramer are linked by disulfide bonds. The redox state of KlADH I could represent the path for modulating its activity and then a regulatory step of glycolysis under hypoxic conditions. It might be hypothesized that KlADH I could represent an important target in redox signaling of Kluyveromyces lactis cell by inhibiting, under oxidative stress, the glycolytic pathway in favor of the pentose-phosphate shunt to restore its reducing potential.

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Gene transfer into hepatocytes is highly desirable for the long-term goal of replacing deficient proteins and correcting metabolic disorders. Bovine herpesvirus 4 (BoHV-4) based vector capability to transduce rat liver cells in vitro and in vivo was assessed. For the in vitro study, a buffalo rat liver cell line was successfully transduced by BoHV-4 and although did not show toxicity, the immediate early two viral gene was transcribed and cells harboring the intact viral genome could be pharmacologically selected, but no viral replication took place. For the in vivo study, adult male rats were inoculated intraportally and intraparenchimally with a BoHV-4 expressing enhanced green fluorescent protein and liver sections were analyzed through fluorescent microscopy. Although the liver parenchyma could not be transduced, the endothelial layer of the liver vasculature showed a robust transgene expression without toxicity. Successful BoHV-4 based vector transduction of primary cultures of rat hepatocytes suggests that extrinsic factors, and not hepatocytes per se, are the cause of such lack of transducibility. The present study serves as a starting point for study of the use of BoHV-4 based vectors to target gene delivery to vascular endothelial cells.

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By computer modelling and protein engineering we have investigated changes in two amino acid residues located in the coenzyme pocket of the yeast Kluyveromyces lactis mitochondrial alcohol dehydrogenase III. These two residues, Gly 225 and Ala 274, were hypothesized to be involved in the enzyme discrimination between NAD(H) and NADP(H). Upon changing Gly 225 to Ala we produced an enzyme (mutant G225A) showing very little difference from the wild-type. On the contrary, change at position 274 of Phe instead of Ala (mutant A274F) caused a significant increase of K(m) values for NAD(P) and for NADPH and even a more marked decrease in catalytic activity. The k(cat)/K(m) rates for NADP(H) were also decreased in this mutant. Enzymes with the double changes at 225 and 274 (mutant G225A-A274F) showed, apart the substantial low K(m) value for NADPH and its high catalytic efficiency, kinetic parameters relative to coenzymes which were not additive over the single substitutions. Surprisingly, enzymes with changes at the two positions reduced efficiently acetaldehyde, displaying a K(m) value 10-fold lower and a catalytic efficiency sevenfold higher with respect to parent or singularly mutated enzymes. None of the engineered enzymes would convert formaldehyde, glutaraldehyde or aromatic aldehydes but all enzymes reduced propionaldehyde and butyraldehyde at relative reaction rates approximately half of that exhibited by acetaldehyde. Interestingly only mutant A274F was able to oxidize methanol almost as well as ethanol. In addition, this mutant was capable to convert secondary and cyclic alcohols, at a rate not detected in the other isoforms. These results are in general agreement with the prediction that increasing the size of amino acids in the proximity of the coenzyme pocket would hamper the accommodation of NADP but discord the increased affinity for NADPH as well as for alcoholic or aldehydic substrates with high steric hindrance.

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