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Highly inflamed and neovascularized corneal graft beds are known as high-risk (HR) environments for transplant survival. One of the primary factors leading to this rejection is reduction in the suppressive function of regulatory T cells (Treg). Our results show that myeloid-derived suppressor cells (MDSC) counteract interleukin-6-mediated Treg dysfunction by expressing interleukin-10. Additionally, MDSC maintain forkhead box P3 stability and their ability to suppress IFN-γ+ Th1 cells. Administering MDSC to HR corneal transplant recipients demonstrates prolonged graft survival via promotion of Treg while concurrently suppressing IFN-γ+ Th1 cells. Moreover, MDSC-mediated donor-specific immune tolerance leads to long-term corneal graft survival as evidenced by the higher survival rate or delayed survival of a second-party C57BL/7 (B6) graft compared to those of third-party C3H grafts observed in contralateral low-risk or HR corneal transplantation of BALB/c recipient mice, respectively. Our study provides compelling preliminary evidence demonstrating the effectiveness of MDSC in preventing Treg dysfunction, significantly improving graft survival in HR corneal transplantation, and showing promising potential for immune tolerance induction.
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Myeloid derived suppressor cells (MDSCs) are a heterogenous population of immature hematopoietic precursors with known immunoregulatory functions. The immunosuppressive role of MDSCs has been highlighted in several inflammatory ophthalmic disorders; however, their therapeutic application in suppressing the immune-mediated changes in dry eye disease (DED) has not been studied. We observed significant reduction in antigen presenting cell (APC) frequencies and their maturation in the presence of MDSCs. Moreover, co-culturing MDSCs with T helper 17 cells (Th17) resulted in reduced Th17 frequencies and their IL-17 expression. On the contrary, MDSCs maintained regulatory T cell frequencies and enhanced their function in-vitro. Furthermore, we delineated the role of interleukin-10 (IL-10) secreted by MDSCs in their immunoregulatory functions. We confirmed these results by flow cytometry analysis and observed that treatment with MDSCs in DED mice effectively suppressed the maturation of APCs, pathogenic Th17 response, and maintained Treg function and significantly ameliorated the disease. The results in this study highlight the potential therapeutic application of MDSCs in treating refractory DED.
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Modelos Animais de Doenças , Síndromes do Olho Seco , Citometria de Fluxo , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides , Linfócitos T Reguladores , Células Th17 , Animais , Células Supressoras Mieloides/imunologia , Síndromes do Olho Seco/imunologia , Síndromes do Olho Seco/metabolismo , Camundongos , Células Th17/imunologia , Linfócitos T Reguladores/imunologia , Células Apresentadoras de Antígenos/imunologia , Feminino , Progressão da Doença , Interleucina-10/metabolismo , Células Cultivadas , Técnicas de CoculturaRESUMO
Corneal transplantation is the most common form of solid tissue grafting, with an approximately 80% to 90% success rate. However, success rates may decline when donor tissues are derived from patients with a history of diabetes mellitus (DM). To evaluate the underlying immunopathologic processes that cause graft rejection, we used streptozotocin-induced type 1 DM (DM1) and transgenic Lepob/ob type 2 DM (DM2) diabetic murine models as donors and nondiabetic BALB/c as recipients. DM resulted in an increased frequency of corneal antigen-presenting cells (APCs) with an acquired immunostimulatory phenotype. Following transplantation, recipients that received either type of diabetic graft showed increased APC migration and T helper type 1 alloreactive cells, impaired functional regulatory T cells, and graft survival. Insulin treatment in streptozotocin-induced diabetic mice led to an increased tolerogenic profile of graft APC, lower T helper type 1 sensitization, and a higher frequency of functional regulatory T cells with high suppressive capacity, reflected in increased graft survival. We conclude that both DM1 and DM2 in donors can impact corneal APC functional phenotype, rendering the tissue more immunogenic and thereby increasing the risk of graft failure.
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Transplante de Córnea , Diabetes Mellitus Experimental , Animais , Camundongos , Diabetes Mellitus Experimental/cirurgia , Diabetes Mellitus Experimental/patologia , Estreptozocina , Córnea , Células Apresentadoras de AntígenosRESUMO
Treatment of isoxazoles with enamines leads to an inverse electron-demand hetero-Diels-Alder reaction that produces substituted pyridines in the presence of TiCl4(THF)2 and titanium powder. The reaction is highly regioselective with only a single isomer of the product observed by GC/MS and tolerant of many common functional groups. The transformation was examined computationally, and it was found that TiCl4 (or a similar Lewis acid) likely acts to catalyze the reaction. After the initial [4 + 2]-cycloaddition, the oxaza-[2.2.1]-bicycle produced likely ring opens before amine loss to give an N-oxide. The pyridine is then obtained after reduction with TiCl4 and titanium powder.
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Isoxazóis , Titânio , Reação de Cicloadição , Pós , Piridinas , EstereoisomerismoRESUMO
Vehicle control systems such as ESC (electronic stability control), MDPS (motor-driven power steering), and ECS (electronically controlled suspension) improve vehicle stability, driver comfort, and safety. Vehicle control systems such as ACC (adaptive cruise control), LKA (lane-keeping assistance), and AEB (autonomous emergency braking) have also been actively studied in recent years as functions that assist drivers to a higher level. These DASs (driver assistance systems) are implemented using vehicle sensors that observe vehicle status and send signals to the ECU (electronic control unit). Therefore, the failure of each system sensor affects the function of the system, which not only causes discomfort to the driver but also increases the risk of accidents. In this paper, we propose a new method to detect and isolate faults in a vehicle control system. The proposed method calculates the constraints and residuals of 12 systems by applying the model-based fault diagnosis method to the sensor of the chassis system. To solve the inaccuracy in detecting and isolating sensor failure, we applied residual sensitivity to a threshold that determines whether faults occur. Moreover, we applied a sensitivity analysis to the parameters semi-correlation table to derive a fault isolation table. To validate the FDI (fault detection and isolation) algorithm developed in this study, fault signals were injected and verified in the HILS (hardware-in-the-loop simulation) environment using an RCP (rapid control prototyping) device.
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PURPOSE: The purpose of this study was to evaluate serial changes in quadriceps and hamstring muscle strength over the first postoperative year in patients who underwent anterior cruciate ligament (ACL) reconstruction with an autologous hamstring tendon graft and to reveal which of these 2 muscles lost more strength and recovered more slowly after autologous hamstring ACL reconstruction. METHODS: Isokinetic muscle strength was measured preoperatively and at 6 months and 1 year postoperatively in 20 patients who underwent ACL reconstruction. The maximal torque (60°/s) and total work (180°/s) of the quadriceps and hamstring were evaluated using an isokinetic testing device. The isokinetic muscle strength and endurance of the injured legs were expressed as percentages of those of the uninjured legs at the same time point. RESULTS: Both quadriceps and hamstring muscle strength at 60°/s and endurance at 180°/s of the injured relative to the uninjured leg was 50% preoperatively. Quadriceps muscle strength and endurance of the injured leg increased to 70% at 6 months and 80% at 1 year postoperatively, whereas hamstring muscle strength and endurance increased to 80% at 6 months and 80% at 1 year. CONCLUSIONS: Knee muscle strength recovered progressively after ACL reconstruction using autologous hamstring tendons but did not fully recover, being about 80% that of the uninjured leg even 1 year after surgery. LEVEL OF EVIDENCE: Level IV, therapeutic case series.
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Reconstrução do Ligamento Cruzado Anterior , Articulação do Joelho/fisiopatologia , Força Muscular/fisiologia , Tendões/transplante , Adolescente , Adulto , Autoenxertos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Resistência Física/fisiologia , Estudos Prospectivos , Adulto JovemRESUMO
PURPOSE: This study was designed to compare the preoperative strengths and endurances of the quadriceps and hamstring muscles in patients with anterior cruciate ligament (ACL) versus posterior cruciate ligament (PCL) tears. METHODS: Quadriceps and hamstring muscle strength and endurance were compared between 20 prospectively enrolled patients with isolated PCL tears and a retrospective, matched control group of 20 patients with isolated ACL tears. The maximal torque (60°/s) and total work (180°/s) of the quadriceps and hamstring were evaluated with an isokinetic testing device. RESULTS: Total work (1,094.4 ± 505.8 J v 797.5 ± 332.7 J, P = .035) and peak torque (129.9 ± 56.2 N â m v 98.2 ± 37.4 N â m, P = .046) of the quadriceps muscle on the involved side were higher in the PCL tear group than in the ACL tear group. However, there were no significant differences between the PCL tear group and ACL tear group in hamstring muscle strength (45.8 ± 42.3 N â m and 46.0 ± 24.4 N â m, respectively; P = .940) and endurance (429.3 ± 238.9 J and 382.4 ± 256.1 J, respectively; P = .574) on the involved side. CONCLUSIONS: The strength and endurance of the quadriceps muscle of the injured limb were greater after PCL tears than after ACL tears. However, there were no significant between-group differences in hamstring muscle strength and endurance on the involved side. LEVEL OF EVIDENCE: Level III, retrospective comparative study.
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Lesões do Ligamento Cruzado Anterior , Força Muscular/fisiologia , Ligamento Cruzado Posterior/lesões , Músculo Quadríceps/fisiopatologia , Adolescente , Adulto , Ligamento Cruzado Anterior/cirurgia , Feminino , Humanos , Traumatismos do Joelho/fisiopatologia , Traumatismos do Joelho/cirurgia , Articulação do Joelho/fisiopatologia , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/fisiopatologia , Ligamento Cruzado Posterior/cirurgia , Estudos Prospectivos , Amplitude de Movimento Articular , Estudos Retrospectivos , Torque , Adulto JovemRESUMO
Ir-catalyzed arene C-H borylations (CHB) of anilines can be highly ortho selective by using a small B2eg2 (eg = ethane-1,2-diol) as the borylating reagent. Unfortunately, the products are prone to decomposition, and transesterification with pinacol is required prior to isolation. This work offers a solution by adjusting the size of the diboron reagent. Based on our evaluation, we conclude that B2bg2 (bg = butane-1,2-diol) achieves an optimal balance between CHB regioselectivity and stability for the borylated products.
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The tear fluid is a readily accessible, potential source for biomarkers of disease and could be used to monitor the ocular response to contact lens (CL) wear or ophthalmic pathologies treated by therapeutic CLs. However, the tear fluid remains largely unexplored as a biomarker source for RNA-based molecular analyses. Using a rabbit model, this study sought to determine whether RNA could be collected from commercial CLs and whether the duration of CL wear would impact RNA recovery. The results were referenced to standardized strips of filtered paper (e.g., Shirmer Strips) placed in the inferior fornix. By performing total RNA isolation, precipitation, and amplification with commercial kits and RT-PCR methods, CLs were found to have no significant differences in RNA concentration and purity compared to Schirmer Strips. The study also identified genes that could be used to normalize RNA levels between tear samples. Of the potential control genes or housekeeping genes, GAPDH was the most stable. This study, which to our knowledge has never been done before, provides a methodology for the detection of RNA and gene expression changes from tear fluid that could be used to monitor or study eye diseases.
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Lentes de Contato , RNA , Lágrimas , Lágrimas/metabolismo , Animais , Coelhos , RNA/isolamento & purificação , RNA/genética , RNA/análiseRESUMO
Fuchs endothelial corneal dystrophy is a heterogenous disease with multifactorial etiology, and genetic, epigenetic, and exogenous factors contributing to its pathogenesis. DNA damage plays a significant role, with ultraviolet-A (UV-A) emerging as a key contributing factor. We investigate the potential application of neuropeptide α-melanocyte stimulating hormone (α-MSH) in mitigating oxidative stress induced endothelial damage. First, we examined the effects of α-MSH on a cultured human corneal endothelial cell line (HCEnC-21T) exposed to hydrogen peroxide (H2O2) induced oxidative DNA damage. We performed immunofluorescence and flow cytometry to assess DNA damage and cell death in the cultured cells. Additionally, we used an established mouse model that utilizes ultraviolet light to induce corneal endothelial cell damage resulting in decreased CEnC number, increased cell size variability, and decreased percentage of hexagonal cells. This endothelial decompensation leads to an increase in corneal thickness. Following UV-A exposure, the mice were systemically treated with α-MSH, either immediately after exposure (early treatment) or beginning two weeks post-exposure (delayed treatment). To evaluate treatment efficacy, we analyzed CEnC density and morphology using in vivo confocal microscopy, and central corneal thickness using anterior segment optical coherence tomography. Our findings demonstrated that α-MSH treatment effectively protects HCEnC-21T from free-radical induced oxidative DNA damage and subsequent cell death. In vivo, α-MSH treatment, mitigated the loss of CEnC density, deterioration of cell morphology and suppression of the resultant corneal swelling. These results underline the potential application of α-MSH as a therapeutic agent for mitigating corneal endothelial damage.
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Dano ao DNA , Modelos Animais de Doenças , Endotélio Corneano , Distrofia Endotelial de Fuchs , Estresse Oxidativo , alfa-MSH , Animais , alfa-MSH/farmacologia , Camundongos , Endotélio Corneano/efeitos dos fármacos , Endotélio Corneano/patologia , Humanos , Distrofia Endotelial de Fuchs/patologia , Distrofia Endotelial de Fuchs/tratamento farmacológico , Dano ao DNA/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Raios Ultravioleta/efeitos adversos , Linhagem Celular , Peróxido de Hidrogênio/farmacologiaRESUMO
A method for modeling high oxidation state catalysts is used on precatalysts with unsymmetrical and symmetrical bidentate ligands to get a more detailed understanding of how changes to ancillary ligands affect the hydroamination of alkynes catalyzed by titanium. To model the electronic donor ability, the ligand donor parameter (LDP) was used, and to model the steric effects, percent buried volume (% Vbur) was employed. For the modeling study, 7 previously unpublished unsymmetrical Ti(XX')(NMe2)2 precatalysts were prepared, where XX' is a chelating ligand with pyrrolyl/indolyl linkages. The rates of these unsymmetrical and 10 previously reported symmetrical precatalysts were used with the model kobs = a + b(LDP)1 + c(LDP)2 + d(% Vbur)1 + e(% Vbur)2, where a-e were found through least-squares refinement. The model suggests that (1) the two attachment points of the bidentate ligand XX' are in different environments on the metal (e.g., axial and equatorial in a trigonal bipyramidal or square pyramidal structure), (2) the position of the unsymmetrical ligand on the metal is determined by the electronics of the ligand rather than the sterics, and (3) that one side of the chelating ligand's electronics strongly influences the rate, while the other side's sterics more strongly influences the rate. From these studies, we were able to generate catalysts fitting to this model with rate constants larger than the fastest symmetrical catalyst tested.
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Purpose: To evaluate whether fibrosis contributes to corneal transplant failure and to determine whether effector CD4+ T cells, the key immune cells in corneal transplant rejection, play a direct role in fibrosis formation. Methods: Allogeneic corneal transplantation was performed in mice. Graft opacity was evaluated by slit-lamp biomicroscopy, and fibrosis was assessed by in vivo confocal microscopy. Expression of alpha-smooth muscle actin (α-SMA) in both accepted and failed grafts was assessed by real-time PCR and immunohistochemistry. Frequencies of graft-infiltrating CD4+ T cells, neutrophils, and macrophages were assessed using flow cytometry. In vitro, MK/T-1 corneal fibroblasts were co-cultured with activated CD4+CD25- effector T cells isolated from corneal transplant recipient mice, and α-SMA expression was quantified by real-time PCR and ELISA. Neutralizing antibody was used to evaluate the role of interferon gamma (IFN-γ) in promoting α-SMA expression. Results: The majority of failed grafts demonstrated clinical signs of fibrosis which became most evident at week 6 after corneal transplantation. Failed grafts showed higher expression of α-SMA as compared to accepted grafts. Flow cytometry analysis showed a significant increase in CD4+ T cells in failed grafts compared to accepted grafts. Co-culture of activated CD4+CD25- effector T cells with corneal fibroblasts led to an increase in α-SMA expression by fibroblasts. Inhibition of IFN-γ in culture significantly suppressed this increase in α-SMA expression as compared to immunoglobulin G control. Conclusions: Fibrosis contributes to graft opacity in corneal transplant failure and is mediated at least in part by effector CD4+ T cells via IFN-γ.
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Doenças da Córnea , Transplante de Córnea , Animais , Camundongos , Linfócitos T CD4-Positivos , Córnea , Anticorpos Neutralizantes , Interferon gamaRESUMO
This study aimed to investigate the effect of a 12-week accelerated rehabilitation exercise program on isokinetic strength and dynamic balance ability of thighs in 20 adult men who underwent anterior cruciate ligament reconstruction (ACLR) or posterior cruciate ligament reconstruction (PCLR) and to analyze intergroup differences in recovery patterns. In this study, we examined 10 patients who underwent ACLR and 10 who underwent PCLR. These patients participated in an accelerated rehabilitation exercise program 5 times weekly for 12 weeks. The participants' isokinetic strength, muscular endurance, and dynamic balance ability of the femoral muscles were measured before and 12 weeks after reconstruction surgery. Isokinetic knee muscle function showed no significant difference between the ACLR and PCLR groups at 60°/sec. Both the groups demonstrated significant increases in muscle strength between the flexors and extensors. However, a between-group difference was noted in knee muscular endurance at 180°/sec, with ACLR patients showing significant differences between extensors and flexors, unlike PCLR patients. Assessment of the dynamic balance ability revealed that overall knee stability did not significantly differ between groups, and both the ACLR and PCLR groups exhibited improved dynamic balance ability. However, significant differences were found in anteroposterior and left-right stabilities. Patients who underwent ACLR had significantly improved anteroposterior and left-right stability, wherever patients who underwent PCLR showed no significant difference. This accelerated rehabilitation exercise program improved the muscle strength and muscular endurance of patients who underwent ACLR and PCLR, suggesting its potential efficacy in recovering dynamic balance ability, particularly after ACLR.
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Biomass production and oil productivity in microalgae culture are the most important key factors for algal biodiesel production. However, proper culture condition for the biomass production of microalgae is different from that for the oil production of microalgae. A study on the biomass production of Tetraselmis suecica using various light intensities and nitrate concentrations as growth factors was carried out to evaluate proper culture conditions in 20-L batch culture. The effect of nitrate depletion on the oil accumulation was also evaluated with two-stage culture. It took 5 days to reach the stationary phase for the cultures of T. suecica on the light intensities of 108.9 and 133.1 µmol m(-2 )s(-1) with biomass of 0.89 and 0.88 g dcw L(-1), respectively. Biomass productions of 1.07 and 1.00 g dcw L(-1) were obtained with the nitrate concentrations of 18.6 and 24.7 mg L(-1), respectively. The two-stage culture increased oil contents from 7.6 to 17.3% (w/w) and contents of C(16)-C(18) fatty acids from 540.2 to 720.5 mg g(-1) oil. The predominant fatty acid was palmitic acid (C(16:0)) in nitrate depletion group, however, oleic acid (C(18:1)) was predominated in nitrate added groups. The two-stage culture enhanced overall oil productivity of 18.7 mg g(-1) day(-1) which is higher than that of 12.2 mg g(-1) day(-1) in single-stage culture.
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Reatores Biológicos/microbiologia , Técnicas de Cultura de Células/métodos , Clorófitas/fisiologia , Nitratos/metabolismo , Óleos/isolamento & purificação , Óleos/metabolismo , Proliferação de Células , Clorófitas/efeitos da radiação , LuzRESUMO
Anti-CD154 blockade-based regimens remain unequaled in prolonging graft survival in various organ transplantation models. Several studies have focused on transplantation tolerance with the anti-CD154 blockade, but none of these studies has investigated the mechanisms associated with its use as the sole treatment in animal models, delaying our understanding of anti-CD154 blockade-mediated immune tolerance. The purpose of this study was to investigate the mechanism underlying the anti-CD154 monoclonal antibody (mAb) blockade in inducing immune tolerance using an intrahepatic murine allogeneic islet transplantation model. Allogeneic BALB/c AnHsd (BALB/c) islets were infused into the liver of diabetic C57BL/6 (B6) mice via the cecal vein. Anti-CD154 mAb (MR1) was administered on -1, 0, 1, 3, 5, and 7 d posttransplantation at 0.5 mg per mouse. We showed that short-term MR1 monotherapy could prolong the allogeneic islet grafts to more than 250 d in the murine intrahepatic islet transplantation model. The second islet grafts transplanted under the kidney capsule of the recipients were protected from rejection. We also found that rejection of same-donor skin grafts transplanted to the tolerant mice was modestly delayed. Using a DEREG mouse model, FoxP3+ regulatory T (Treg) cells were shown to play important roles in transplantation tolerance. In mixed lymphocyte reactions, Treg cells from the tolerant mice showed more potency in suppressing BALB/c splenocyte-stimulated Teff cell proliferation than those from naïve mice. In this study, we demonstrated for the first time that a short-term anti-CD154 mAb single treatment could induce FoxP3+ Treg cell-mediated immune tolerance in the intrahepatic murine allogeneic islet transplantation model.
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Anticorpos Monoclonais/farmacologia , Rejeição de Enxerto/tratamento farmacológico , Tolerância Imunológica/efeitos dos fármacos , Transplante das Ilhotas Pancreáticas/imunologia , Linfócitos T Reguladores/efeitos dos fármacos , Animais , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/efeitos dos fármacos , Sobrevivência de Enxerto/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Transplante de Pele/métodos , Linfócitos T Reguladores/imunologia , Tolerância ao Transplante , Transplante Homólogo/métodosRESUMO
This study investigated the pathogenesis of periodontitis and the role of nucleotide-binding oligomerization domain-like receptor protein 10 (NLRP10). The human oral epithelial cell line HOK-16B was infected with two periodontal pathogens, Tannerella forsythia and Fusobacterium nucleatum, at various MOIs. RT-PCR and immunoblotting demonstrated that infection increased mRNA and protein expression of NLRP10, respectively. The siRNA-mediated NLRP10 knockdown significantly reduced IL-1α expression and secretion. Both bacteria induced phosphorylation of ERK, JNK and p38 MAP kinases in HOK-16B cells. NLRP10 knockdown impaired ERK phosphorylation only. ERK inhibition significantly decreased the expression of T. forsythia- and F. nucleatum-induced IL-1α. Our data suggest that NLRP10 is involved in activating the ERK signalling pathway in HOK-16B cells infected with T. forsythia and F. nucleatum. This pathway likely augments the pro-inflammatory cytokine IL-1α levels, which may play a critical role in periodontitis.
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Proteínas de Transporte/metabolismo , Células Epiteliais/imunologia , Infecções por Fusobacterium/imunologia , Fusobacterium nucleatum/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Periodontite/imunologia , Tannerella forsythia/imunologia , Proteínas Adaptadoras de Transdução de Sinal , Proteínas Reguladoras de Apoptose , Proteínas de Transporte/genética , Linhagem Celular , Células Epiteliais/microbiologia , Regulação da Expressão Gênica , Humanos , Interleucina-1alfa/genética , Interleucina-1alfa/metabolismo , Sistema de Sinalização das MAP Quinases , Boca/citologia , RNA Interferente Pequeno/genéticaRESUMO
PURPOSE: The purpose of this study was to evaluate the quadriceps and hamstring muscle strength and hamstring-to-quadriceps (HQ) ratio, as well as the relationships of these parameters with dynamic balance, in patients with anterior cruciate ligament (ACL) rupture. MATERIALS AND METHODS: We compared 25 patients diagnosed with chronic unilateral ACL tears and 25 age-matched healthy volunteers. The maximal torque of the quadriceps and hamstring and dynamic balance were measured. RESULTS: Although the isokinetic maximal peak torques were about 50% lower in the quadriceps (57%, p<0.001) and hamstring (56%, p=0.001) muscles in the chronic ACL tear group than in the control group, their HQ ratios were similar (56%±17% vs. 58%±6%, p=0.591). HQ ratio was significantly correlated with anterior-posterior stability index (r=-0.511, p=0.021) and overall stability index (r=-0.476, p=0.034) in control group, but these correlations were not observed in chronic ACL tear group. CONCLUSIONS: Thigh muscle strength was about 50% lower in the chronic ACL tear group than in the control group, but the HQ ratio was similar. The dynamic balance of the knee was not influenced by thigh muscle strength but was influenced by HQ ratio in healthy young individuals. However, HQ ratio was not correlated with dynamic knee balance in chronic ACL tear patients.