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1.
J Arthroplasty ; 38(9): 1726-1733.e4, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-36924858

RESUMO

BACKGROUND: The rate of using robotic-assisted total knee arthroplasty (RA-TKA) has increased markedly. Understanding how patients view the role of robotics during total knee arthroplasty (TKA) informs shared decision making and facilitate efforts to appropriately educate patients regarding the risks and benefits of robotic assistance. METHODS: A self-administered questionnaire was completed by 440 potential TKA patients at the time of their surgery scheduling. Participants answered 25 questions regarding RA-TKA, socioeconomic factors, and their willingness to pay (WTP) for RA-TKA. Logistic regressions were used to determine if population characteristics and surgeon preferences influenced the patients' perceptions of RA-TKA. RESULTS: There were 39.7% of respondents who said that they had no knowledge regarding RA-TKA. Only 40.7% of participants had expressed a desire for RA-TKA to be used. There were 8.7% who were WTP extra for the use of RA-TKA. Participants believed that the main 3 benefits of RA-TKA compared to conventional methods were: more accurate implant placement (56.2%); better results (49.0%); and faster recovery (32.1%). The main 3 patient concerns were harm from malfunction (55.2%), reduced surgeon role in the procedure (48.1%), and lack of supportive research (28.3%). Surgeon preference of RA-TKA was associated with patient's willingness to have RA-TKA (odds ratio 4.60, confidence interval 2.98-7.81, P < .001), and with WTP extra for RA-TKA (odds ratio 2.05, confidence interval: 1.01-4.26, P = .049). CONCLUSION: Patient knowledge regarding RA-TKA is limited. Nonpeer-reviewed online information may make prospective TKA candidates vulnerable to misinformation and aggressive advertising. The challenge for orthopaedic surgeons is to re-establish control and reliably educate patients about the proven advantages and disadvantages of this emerging technology.


Assuntos
Artroplastia do Joelho , Osteoartrite do Joelho , Procedimentos Cirúrgicos Robóticos , Humanos , Artroplastia do Joelho/métodos , Articulação do Joelho/cirurgia , Osteoartrite do Joelho/cirurgia , Estudos Prospectivos , Motivação , Procedimentos Cirúrgicos Robóticos/métodos
2.
JBJS Case Connect ; 12(2)2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-37440519

RESUMO

CASE: Medial collateral ligament (MCL) injuries after total knee arthroplasty (TKA) are a rare occurrence. Traditional treatment options include repair and revision to a constrained prosthesis. We present a case of an 80-year-old man who opted for MCL reconstruction in the setting of previous TKA due to late MCL injury from a skiing accident. The presentation, operative techniques, and outcomes of this treatment option are presented. CONCLUSIONS: MCL reconstruction in the setting of TKA can lead to a successful and satisfactory outcome without the need for revision TKA.


Assuntos
Artroplastia do Joelho , Instabilidade Articular , Ligamento Colateral Médio do Joelho , Masculino , Humanos , Idoso de 80 Anos ou mais , Artroplastia do Joelho/métodos , Articulação do Joelho/cirurgia , Resultado do Tratamento , Ligamento Colateral Médio do Joelho/cirurgia , Ligamento Colateral Médio do Joelho/lesões , Instabilidade Articular/etiologia , Instabilidade Articular/cirurgia , Estudos Retrospectivos
3.
J Arthroplasty ; 36(6): 1857-1863, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33579631

RESUMO

BACKGROUND: Optimizing surgical trays to improve operating room efficiency and reduce costs in instrument processing is an under-appreciated strategy for cost containment. This study aimed to assess the economic impact of instrument tray optimization in total joint arthroplasty. METHODS: Thirty-five randomly selected elective primary total knee arthroplasty and total hip arthroplasty performed by 4 fellowship-trained surgeons were analyzed. Type and number of instruments used as well as timing of different steps in the sterilization process were recorded by an independent observer. Using Lean methodology, surgeons identified redundant or underutilized instruments and agreed upon the fewest number needed for each tray. Instrument utilization rates and processing time were analyzed before and after tray modifications. Annual cost savings were calculated based on a processing factor of $0.59-$11.52 per instrument. RESULTS: Only 45.5% of instruments opened for total knee arthroplasty were utilized. After optimization, 28 of 87 (32.2%) instruments were removed and the remainder could be stored in one tray. Mean set-up time decreased from 20.7 to 14.2 minutes, while 40-75 minutes were saved during the sterilization process. For total hip arthroplasty, only 36.0% of instruments were utilized. Using Lean methods, 46 of 112 (41.1%) instruments were removed and tray count was reduced to 2 trays. Mean set-up time decreased from 27.9 to 18.6 minutes, while 45-150 minutes were saved during processing. Average annual savings amounted to $281,298.05. CONCLUSION: Lean methodology can be used to eliminate redundant or underutilized instruments in total joint arthroplasty, improving surgical efficiency and generating substantial cost savings.


Assuntos
Salas Cirúrgicas , Instrumentos Cirúrgicos , Artroplastia , Redução de Custos , Humanos , Esterilização
5.
JBJS Case Connect ; 10(2): e19.00489, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32649150

RESUMO

CASE: A 67-year-old immunocompromised woman presented with 2 weeks of atraumatic knee pain 4 months after a primary total knee arthroplasty. The patient had a history of consuming unpasteurized dairy products and horse rearing. The patient underwent a successful irrigation, debridement, polyethylene liner exchange, and adjunct antibiotic therapy for a periprosthetic joint infection (PJI). Intraoperative cultures grew Listeria monocytogenes. CONCLUSION: Immmunocompromised patients are at increased risks for PJI. Those patients who engage in rearing animals and consuming unpasteurized dairy products are at increased risk for atypical infections. It is vital to recognize patients who are at increased risk for infection and council them on risk aversion behavior.


Assuntos
Antibacterianos/administração & dosagem , Artrite Infecciosa/microbiologia , Artroplastia do Joelho/efeitos adversos , Listeria monocytogenes/isolamento & purificação , Infecções Relacionadas à Prótese/microbiologia , Idoso , Artrite Infecciosa/tratamento farmacológico , Artrite Infecciosa/cirurgia , Feminino , Humanos , Infecções Relacionadas à Prótese/tratamento farmacológico , Infecções Relacionadas à Prótese/cirurgia
6.
Respir Res ; 21(1): 108, 2020 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-32381012

RESUMO

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a rapidly progressing disease with challenging management. To find novel effective therapies, better preclinical models are needed for the screening of anti-fibrotic compounds. Activated fibroblasts drive fibrogenesis and are the main cells responsible for the accumulation of extracellular matrix (ECM). Here, a prolonged Scar-in-a-Jar assay was combined with clinically validated biochemical markers of ECM synthesis to evaluate ECM synthesis over time. To validate the model as a drug screening tool for novel anti-fibrotic compounds, two approved compounds for IPF, nintedanib and pirfenidone, and a compound in development, omipalisib, were tested. METHODS: Primary human lung fibroblasts from healthy donors were cultured for 12 days in the presence of ficoll and were stimulated with TGF-ß1 with or without treatment with an ALK5/TGF-ß1 receptor kinase inhibitor (ALK5i), nintedanib, pirfenidone or the mTOR/PI3K inhibitor omipalisib (GSK2126458). Biomarkers of ECM synthesis were evaluated over time in cell supernatants using ELISAs to assess type I, III, IV, V and VI collagen formation (PRO-C1, PRO-C3, PRO-C4, PRO-C5, PRO-C6), fibronectin (FBN-C) deposition and α-smooth muscle actin (α-SMA) expression. RESULTS: TGF-ß1 induced synthesis of PRO-C1, PRO-C6 and FBN-C as compared with unstimulated fibroblasts at all timepoints, while PRO-C3 and α-SMA levels were not elevated until day 8. Elevated biomarkers were reduced by suppressing TGF-ß1 signalling with ALK5i. Nintedanib and omipalisib were able to reduce all biomarkers induced by TGF-ß1 in a concentration dependent manner, while pirfenidone had no effect on α-SMA. CONCLUSIONS: TGF-ß1 stimulated synthesis of type I, III and VI collagen, fibronectin and α-SMA but not type IV or V collagen. Synthesis was increased over time, although temporal profiles differed, and was modulated pharmacologically by ALK5i, nintedanib, pirfenidone and omipalisib. This prolonged 12-day Scar-in-a-Jar assay utilising biochemical markers of ECM synthesis provides a useful screening tool for novel anti-fibrotic compounds.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Cicatriz/induzido quimicamente , Cicatriz/metabolismo , Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Biomarcadores/metabolismo , Células Cultivadas , Cicatriz/tratamento farmacológico , Colágeno/antagonistas & inibidores , Colágeno/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Matriz Extracelular/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibronectinas/antagonistas & inibidores , Fibronectinas/metabolismo , Fibrose/induzido quimicamente , Fibrose/tratamento farmacológico , Fibrose/metabolismo , Humanos , Indóis/antagonistas & inibidores , Indóis/metabolismo , Inibidores de Proteínas Quinases/uso terapêutico , Piridonas/antagonistas & inibidores , Piridonas/metabolismo , Fator de Crescimento Transformador beta1/toxicidade
7.
J Arthroplasty ; 35(5): 1228-1232, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32115328

RESUMO

BACKGROUND: Controlling postoperative pain and reducing opioid requirements after total knee arthroplasty remain a challenge, particularly in an era stressing rapid recovery protocols and early discharge. A single-shot adductor canal blockade (ACB) has been shown to be effective in decreasing postoperative pain. The purpose of the present study is to compare the efficacy of an anesthesiologist administered ACB and a surgeon administered intraoperative ACB. METHODS: Patients undergoing primary total knee arthroplasty were prospectively randomized to receive either an anesthesiologist administered (group 1) or surgeon administered (group 2) ACB using 15 mL of ropivacaine 0.5%. Primary outcomes were pain visual analog scale, range of motion, and opioid consumption. RESULTS: Thirty-four patients were randomized to group 1 and 29 to group 2. Opioid equivalents consumed were equal on postoperative day (POD) 0, 1, and 2. Patients in group 1 had statistically less pain on POD 0, but this did not reach clinical significance and there was no difference in pain on POD 1 or 2. Patients in group 1 had significantly increased active flexion POD 1, but there was no difference in active flexion on POD 0 or 6 weeks postop. There was no difference in patient satisfaction with pain control or short-term functional outcomes. CONCLUSION: Surgeon administered ACB is not inferior to anesthesiologist administered ACB with respect to pain, opioid consumption, range of motion, patient satisfaction, or short-term functional outcomes. Surgeon administered ACB is an effective alternative to anesthesiologist administered ACB.


Assuntos
Bloqueio Nervoso , Cirurgiões , Anestesiologistas , Anestésicos Locais , Humanos , Dor Pós-Operatória/prevenção & controle , Estudos Prospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto
8.
Sci Rep ; 9(1): 11898, 2019 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-31417162

RESUMO

Myzus persicae is a major pest of many crops including canola and Brassica vegetables, partly because it vectors plant viruses. Previously it has been reported that double-stranded RNA delivered to aphids by injection, artificial diet or transgenic plants has knocked down target genes and caused phenotypic effects. While these studies suggest that RNA interference (RNAi) might be used to suppress aphid populations, none have shown effects sufficient for field control. The current study analyses the efficacy of dsRNA directed against previously reported gene-targets on Green peach aphid (Myzus persicae) strains. No silencing effect was observed when dsRNA was delivered in artificial diet with or without transfection reagents. dsRNA produced in planta also failed to induce significant RNAi in M. persicae. Transcriptome analyses of the midgut suggested other potential targets including the Ferritin heavy chain transcripts, but they also could not be knocked down with dsRNA. Here we show that dsRNA is rapidly degraded by midgut secretions of Myzus persicae. Analysis of the transcriptome of the M. persicae midgut revealed that an ortholog of RNases from other insects was abundant.


Assuntos
Afídeos/enzimologia , Sistema Digestório/enzimologia , Endonucleases/metabolismo , Espaço Extracelular/enzimologia , Interferência de RNA , Administração Oral , Sequência de Aminoácidos , Animais , Arabidopsis/genética , Peso Corporal , Dieta , Endonucleases/química , Ferritinas/genética , Filogenia , Plantas Geneticamente Modificadas , RNA de Cadeia Dupla/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
9.
Proc Natl Acad Sci U S A ; 116(21): 10424-10429, 2019 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-31064874

RESUMO

Insecticide resistance is a paradigm of microevolution, and insecticides are responsible for the strongest cases of recent selection in the genome of Drosophila melanogaster Here we use a naïve population and a novel insecticide class to examine the ab initio genetic architecture of a potential selective response. Genome-wide association studies (GWAS) of chlorantraniliprole susceptibility reveal variation in a gene of major effect, Stretchin Myosin light chain kinase (Strn-Mlck), which we validate with linkage mapping and transgenic manipulation of gene expression. We propose that allelic variation in Strn-Mlck alters sensitivity to the calcium depletion attributable to chlorantraniliprole's mode of action. GWAS also reveal a network of genes involved in neuromuscular biology. In contrast, phenotype to transcriptome associations identify differences in constitutive levels of multiple transcripts regulated by cnc, the homolog of mammalian Nrf2. This suggests that genetic variation acts in trans to regulate multiple metabolic enzymes in this pathway. The most outstanding association is with the transcription level of Cyp12d1 which is also affected in cis by copy number variation. Transgenic overexpression of Cyp12d1 reduces susceptibility to both chlorantraniliprole and the closely related insecticide cyantraniliprole. This systems genetics study reveals multiple allelic variants segregating at intermediate frequency in a population that is completely naïve to this new insecticide chemistry and it foreshadows a selective response among natural populations to these chemicals.


Assuntos
Variações do Número de Cópias de DNA/genética , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Transativadores/genética , Alelos , Animais , Animais Geneticamente Modificados , Mapeamento Cromossômico/métodos , Variações do Número de Cópias de DNA/efeitos dos fármacos , Proteínas de Drosophila/genética , Estudo de Associação Genômica Ampla/métodos , Quinase de Cadeia Leve de Miosina/genética , Fenótipo , Pirazóis/farmacologia , Sobrevivência , Transcriptoma/efeitos dos fármacos , Transcriptoma/genética , ortoaminobenzoatos/farmacologia
10.
J Arthroplasty ; 34(8): 1640-1645, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31084971

RESUMO

BACKGROUND: Multiple studies have demonstrated that ketamine, a glutamate receptor blocker, may decrease postoperative pain in abdominal and orthopedic surgeries. However, its role with spinal anesthesia and total knee arthroplasty (TKA) remains unknown. The purpose of this study is to determine the efficacy of subanesthetic dosing of ketamine during TKA on postoperative pain and narcotic consumption. METHODS: In this prospective, randomized, double-blinded clinical trial, we enrolled 91 patients undergoing primary TKA with spinal anesthesia in a single institution from 2017 to 2018. Patients were randomized to receive intraoperative ketamine infusion at a rate of 6 mcg/kg/min for 75 minutes or a saline placebo. All patients received spinal anesthesia and otherwise identical surgical approaches, pain management, and rehabilitation protocols. Patient-reported visual analog pain scores were calculated preoperatively, postoperative days (POD) 0-7, and 2 weeks. Narcotic consumption was evaluated on POD 0 and 1. RESULTS: There was no difference in average pain between ketamine and placebo at all time points except for at PODs 1 (45 vs 56, P = .041) and 4 (39 vs 49, P = .040). For least pain experienced, patients administered with ketamine experienced a reduction in pain only at POD 4 (22 vs 35, P = .011). There was no difference in maximum pain cohorts at all time points of the study or in-hospital morphine equivalents between the 2 cohorts. CONCLUSION: As part of multimodal pain management protocol, intraoperative ketamine does not result in a clinically significant improvement in pain and narcotic consumption following TKA.


Assuntos
Analgesia Controlada pelo Paciente/métodos , Analgésicos Opioides/administração & dosagem , Artroplastia do Joelho , Ketamina/uso terapêutico , Entorpecentes/uso terapêutico , Idoso , Raquianestesia/métodos , Método Duplo-Cego , Feminino , Humanos , Período Intraoperatório , Masculino , Pessoa de Meia-Idade , Morfina/uso terapêutico , Manejo da Dor/métodos , Medição da Dor , Dor Pós-Operatória/tratamento farmacológico , Período Pós-Operatório , Estudos Prospectivos
11.
J Arthroplasty ; 34(9): 1963-1968, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31104838

RESUMO

BACKGROUND: Currently, the decision to resurface the patella is often made irrespective of the presence of patellar arthritis. The purpose of this study is to utilize the existing literature to assess cost-utility of routinely vs selectively resurfacing the patella. METHODS: Prospective randomized studies of patella resurfacing vs non-resurfacing in total knee arthroplasty (TKA) were identified through literature review. Data from these studies represented probabilities of varied outcomes following TKA dependent upon patella resurfacing. Using previously validated utility scores from the McKnee modified Health Utilities Index, endpoint utility values were provided for each potential outcome. RESULTS: Literature review yielded a total of 14 studies with 3,562 patients receiving 3,823 TKAs, of which 1,873 (49.0%) patellae were resurfaced. Persistent postoperative anterior knee pain occurred in 20.9% vs 13.2% (P < .001) and patella reoperation was performed in 3.7% vs 1.6% (P < .001) of unresurfaced and resurfaced patella, respectively. In studies excluding those with arthritic patellae, the incidence of anterior knee pain was equivalent between groups and reoperation decreased to 1.2% vs 0% (P = .06). Patella resurfacing provided marginally improved quality-adjusted life-years (QALY) for both selective and indiscriminate patella resurfacing. When including all studies, the incremental cost per QALY was $3,032. However, when analyzing only those studies with nonarthritic patellae, the incremental cost per QALY to resurface the patella increased to $183,584. CONCLUSION: Patellar resurfacing remains a controversial issue in TKA. Utilizing data from new prospective randomized studies, this analysis finds that routinely resurfacing arthritis-free patellae in TKA are not cost-effective.


Assuntos
Artroplastia do Joelho/economia , Artroplastia do Joelho/métodos , Articulação do Joelho/cirurgia , Osteoartrite do Joelho/cirurgia , Patela/cirurgia , Análise Custo-Benefício , Árvores de Decisões , Humanos , Osteoartrite do Joelho/economia , Probabilidade , Estudos Prospectivos , Anos de Vida Ajustados por Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Reoperação , Resultado do Tratamento
12.
CRISPR J ; 2: 31-40, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-31021235

RESUMO

Genome editing is a tool that has many applications, including the validation of potential drug targets. However, performing genome editing in low-passage primary human cells with the greatest physiological relevance is notoriously difficult. High editing efficiency is desired because it enables gene knockouts (KO) to be generated in bulk cellular populations and circumvents the problem of having to generate clonal cell isolates. Here, we describe a single-step workflow enabling >90% KO generation in primary human lung fibroblasts via CRISPR ribonucleoprotein delivery in the absence of antibiotic selection or clonal expansion. As proof of concept, we edited two SMAD family members and demonstrated that in response to transforming growth factor beta, SMAD3, but not SMAD2, is critical for deposition of type I collagen in the fibrotic response. The optimization of this workflow can be readily transferred to other primary cell types.


Assuntos
Edição de Genes/métodos , Técnicas de Inativação de Genes/métodos , Cultura Primária de Células/métodos , Sistemas CRISPR-Cas/genética , Técnicas de Cultura de Células/métodos , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Fibroblastos/metabolismo , Engenharia Genética/métodos , Vetores Genéticos , Humanos , Pulmão/patologia , Proteínas Smad/genética , Proteínas Smad/metabolismo
13.
Nat Ecol Evol ; 3(4): 647-656, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30886368

RESUMO

Imidacloprid, the world's most used insecticide, has caused considerable controversy due to harmful effects on non-pest species and increasing evidence showing that insecticides have become the primary selective force in many insect species. The genetic response to insecticides is heterogeneous across populations and environments, leading to more complex patterns of genetic variation than previously thought. This motivated the investigation of imidacloprid resistance at different temperatures in natural populations of Drosophila melanogaster originating from four climate extremes replicated across two continents. Population and quantitative genomic analysis, supported by functional tests, have revealed a mixed genetic architecture to resistance involving major genes (Paramyosin and Nicotinic-Acetylcholine Receptor Alpha 3) and polygenes with a major trade-off with thermotolerance. Reduced genetic differentiation at resistance-associated loci indicated enhanced gene flow at these loci. Resistance alleles showed stronger evidence of positive selection in temperate populations compared to tropical populations in which chromosomal inversions In(2 L)t, In(3 R)Mo and In(3 R)Payne harbour susceptibility alleles. Polygenic architecture and ecological factors should be considered when developing sustainable management strategies for both pest and beneficial insects.


Assuntos
Drosophila melanogaster/fisiologia , Resistência a Inseticidas/fisiologia , Inseticidas , Neonicotinoides , Nitrocompostos , Termotolerância , Animais , Clima , Feminino , Estudo de Associação Genômica Ampla , Receptores Nicotínicos/genética , Tropomiosina/genética
14.
Nat Commun ; 10(1): 6, 2019 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-30602778

RESUMO

Myofibroblasts are the key effector cells responsible for excessive extracellular matrix deposition in multiple fibrotic conditions, including idiopathic pulmonary fibrosis (IPF). The PI3K/Akt/mTOR axis has been implicated in fibrosis, with pan-PI3K/mTOR inhibition currently under clinical evaluation in IPF. Here we demonstrate that rapamycin-insensitive mTORC1 signaling via 4E-BP1 is a critical pathway for TGF-ß1 stimulated collagen synthesis in human lung fibroblasts, whereas canonical PI3K/Akt signaling is not required. The importance of mTORC1 signaling was confirmed by CRISPR-Cas9 gene editing in normal and IPF fibroblasts, as well as in lung cancer-associated fibroblasts, dermal fibroblasts and hepatic stellate cells. The inhibitory effect of ATP-competitive mTOR inhibition extended to other matrisome proteins implicated in the development of fibrosis and human disease relevance was demonstrated in live precision-cut IPF lung slices. Our data demonstrate that the mTORC1/4E-BP1 axis represents a critical signaling node during fibrogenesis with potential implications for the development of novel anti-fibrotic strategies.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Colágeno/biossíntese , Fibroblastos/metabolismo , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Fosfoproteínas/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Proteínas de Ciclo Celular , Linhagem Celular , Humanos , Fibrose Pulmonar Idiopática/etiologia , Fosfatidilinositol 3-Quinases/metabolismo , Transdução de Sinais , Sirolimo , Serina-Treonina Quinases TOR/metabolismo
15.
BMC Biomed Eng ; 1: 14, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32903343

RESUMO

BACKGROUND: Excessive extracellular matrix (ECM) deposition is a hallmark feature in fibrosis and tissue remodelling diseases. Typically, mesenchymal cells will produce collagens under standard 2D cell culture conditions, however these do not assemble into fibrils. Existing assays for measuring ECM production are often low throughput and not disease relevant. Here we describe a robust, high content, pseudo-3D phenotypic assay to quantify mature fibrillar collagen deposition which is both physiologically relevant and amenable to high throughput compound screening. Using pulmonary fibroblasts derived from patients with idiopathic pulmonary fibrosis (IPF), we developed the 'scar-in-a-jar' assay into a medium-throughput phenotypic assay to robustly quantify collagen type I deposition and other extracellular matrix (ECM) proteins over 72 h. RESULTS: This assay utilises macromolecular crowding to induce an excluded volume effect and enhance enzyme activity, which in combination with TGF-ß1 stimulation significantly accelerates ECM production. Collagen type I is upregulated approximately 5-fold with a negligible effect on cell number. We demonstrate the robustness of the assay achieving a Z prime of approximately 0.5, and % coefficient of variance (CV) of < 5 for the assay controls SB-525334 (ALK5 inhibitor) and CZ415 (mTOR inhibitor). This assay has been used to confirm the potency of a number of potential anti-fibrotic agents. Active compounds from the 'scar-in-a-jar' assay can be further validated for other markers of ECM deposition and fibroblast activation such as collagen type IV and α-smooth muscle actin exhibiting a 4-fold and 3-fold assay window respectively. CONCLUSION: In conclusion, we have developed 'scar -in-a-jar is' into a robust disease-relevant medium-throughput in vitro assay to accurately quantify ECM deposition. This assay may enable iterative compound profiling for IPF and other fibroproliferative and tissue remodelling diseases.

16.
Elife ; 72018 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-29345617

RESUMO

Malignant mesothelioma (MM) is poorly responsive to systemic cytotoxic chemotherapy and invariably fatal. Here we describe a screen of 94 drugs in 15 exome-sequenced MM lines and the discovery of a subset defined by loss of function of the nuclear deubiquitinase BRCA associated protein-1 (BAP1) that demonstrate heightened sensitivity to TRAIL (tumour necrosis factor-related apoptosis-inducing ligand). This association is observed across human early passage MM cultures, mouse xenografts and human tumour explants. We demonstrate that BAP1 deubiquitinase activity and its association with ASXL1 to form the Polycomb repressive deubiquitinase complex (PR-DUB) impacts TRAIL sensitivity implicating transcriptional modulation as an underlying mechanism. Death receptor agonists are well-tolerated anti-cancer agents demonstrating limited therapeutic benefit in trials without a targeting biomarker. We identify BAP1 loss-of-function mutations, which are frequent in MM, as a potential genomic stratification tool for TRAIL sensitivity with immediate and actionable therapeutic implications.


Assuntos
Neoplasias Pulmonares/fisiopatologia , Mesotelioma/fisiopatologia , Proteínas Repressoras/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina Tiolesterase/metabolismo , Animais , Linhagem Celular Tumoral , Humanos , Mesotelioma Maligno , Camundongos
17.
Genetics ; 207(3): 1181-1193, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28935691

RESUMO

Insecticide resistance is considered a classic model of microevolution, where a strong selective agent is applied to a large natural population, resulting in a change in frequency of alleles that confer resistance. While many insecticide resistance variants have been characterized at the gene level, they are typically single genes of large effect identified in highly resistant pest species. In contrast, multiple variants have been implicated in DDT resistance in Drosophila melanogaster; however, only the Cyp6g1 locus has previously been shown to be relevant to field populations. Here we use genome-wide association studies (GWAS) to identify DDT-associated polygenes and use selective sweep analyses to assess their adaptive significance. We identify and verify two candidate DDT resistance loci. A largely uncharacterized gene, CG10737, has a function in muscles that ameliorates the effects of DDT, while a putative detoxifying P450, Cyp6w1, shows compelling evidence of positive selection.


Assuntos
DDT/toxicidade , Drosophila melanogaster/genética , Loci Gênicos , Resistência a Inseticidas/genética , Praguicidas/toxicidade , Animais , Sistema Enzimático do Citocromo P-450/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/efeitos dos fármacos , Genoma de Inseto , Seleção Genética
18.
J Bone Joint Surg Am ; 99(16): 1337-1344, 2017 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-28816893

RESUMO

BACKGROUND: Intra-articular bupivacaine hydrochloride (HCl) infusion catheters and periarticular injections of liposomal bupivacaine are often used as postoperative local anesthetics. The purpose of this study was to compare the efficacies of these local anesthetics following total knee arthroplasty. METHODS: This study was a superiority trial with a randomized, controlled, double-blinded design. Patients were randomly assigned to either delivery of bupivacaine HCl by the ON-Q* Pain Relief System pump (n = 96) or by an injection of Exparel (liposomal bupivacaine) (n = 104). The primary outcome of this study was cumulative narcotic consumption on postoperative days 0 through 3. Narcotic consumption data were collected retrospectively from in-hospital records while patients were in the hospital. Following discharge, narcotic consumption data were gathered from patient surveys, as were secondary outcomes measures. RESULTS: We did not identify greater narcotic use in the ON-Q* group compared with the Exparel group (p = 0.641). The mean difference between the groups was 0.5 morphine equivalent (95% confidence interval [CI] = -1.7 to +2.8), with the ON-Q* group consuming an average 10.4 morphine equivalents (95% CI = 8.7 to 12.0) compared with 10.9 (95% CI = 9.3 to 12.5) in the Exparel group. There were no significant differences between groups with regard to any of the secondary measures of pain with the exception of pain while walking and pain with physical therapy (p = 0.019 and p = 0.010, respectively), both of which showed an approximately 1-point difference in favor of the ON-Q* group on a visual analog scale (VAS). There were also no differences in the postoperative side effects, including nausea, constipation, or vomiting, or in the rates of study-related complications, patient satisfaction, or length of hospital stay. CONCLUSIONS: Exparel did not have superior efficacy compared with the ON-Q* Pain Relief System as reflected by narcotic consumption, our primary outcome. There were small significant differences, in favor of the ON-Q* group, in 2 secondary measures of pain during activity, but these approximately 1-point VAS differences are unlikely to be clinically relevant. The choice of a local anesthetic modality should be based on a combination of safety, convenience, and cost considerations. LEVEL OF EVIDENCE: Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.


Assuntos
Anestesia Local/métodos , Anestésicos Locais/administração & dosagem , Artroplastia do Joelho , Bupivacaína/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Bombas de Infusão , Adulto , Idoso , Idoso de 80 Anos ou mais , Cateterismo/métodos , Método Duplo-Cego , Feminino , Humanos , Injeções Intra-Articulares , Tempo de Internação , Modelos Lineares , Lipossomos , Masculino , Pessoa de Meia-Idade , Manejo da Dor , Medição da Dor , Satisfação do Paciente
19.
J Arthroplasty ; 31(7): 1583-8, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26869061

RESUMO

BACKGROUND: Vascular complications during total knee arthroplasty (TKA) are uncommon but potentially devastating. We evaluated cases of vascular complication during TKA in our high-volume, community hospital system. METHODS: Patients were identified by cross-referencing billing codes for TKA with diagnosis codes for vascular complication during the same hospital stay between January 1, 2010 and December 31, 2014. Clinical and radiographic data, time to diagnosis, intervention, and outcomes were collected. RESULTS: We identified 13 vascular complications in 9951 TKAs (0.13%). Average age was 66.2 years (95% CI: 5.55; range: 54.1-87.9), 12 (92.3%) were female, average body mass index was 32.3 (5.17; 20-50), and average Charlson Comorbidity Index was 4.08 (1.03; 2-7). Black females (relative risk = 18.33, 95% CI: 6.20-54.22) were at particularly high risk. Preoperatively, 6 knees exhibited varus coronal malalignment and 2 valgus malalignment (only 1 >15°). None had flexion contracture >10°. Four knees exhibited vascular calcifications on preoperative radiographs. Twelve were diagnosed and treated the same day as index TKA and 1 on postoperative day 2. All underwent interventions: 9 stents, 2 endarterectomies, 1 thrombectomy, and 1 bypass. One patient sustained a peroneal nerve injury; 3 had persistent stiffness postoperatively that improved after manipulation. There were no revision surgeries, further vascular intervention, compartment syndrome, periprosthetic joint infection, amputation, or mortality. CONCLUSION: Incidence of vascular complications at our community-based hospital system is in line with previous reports. Black race and female gender were significant risk factors. Although outcomes were remarkable for a high rate of stiffness and one peroneal neuropathy, the devastating complications of amputation, compartment syndrome, periprosthetic joint infection, or early mortality were not observed.


Assuntos
Artrite Infecciosa/etiologia , Artroplastia do Joelho/efeitos adversos , Síndromes Compartimentais/etiologia , Articulação do Joelho/cirurgia , Doenças Vasculares/etiologia , Idoso , Serviços de Saúde Comunitária , Feminino , Hospitais Comunitários , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Reoperação/estatística & dados numéricos , Fatores de Risco , Tempo para o Tratamento
20.
Surg Technol Int ; 27: 225-32, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26680402

RESUMO

PURPOSE: Accurate alignment and balanced flexion and extension gaps are critical elements in achieving a successful outcome following total knee arthroplasty (TKA). The ability to make accurate and precise bone cuts is essential in the creation of balanced gaps. We sought to determine if one type of modern-day standard surgical instrument using an intramedullary rod and posterior referencing produces accurate and precise distal and posterior femoral bone resections. MATERIALS AND METHODS: Seventy-five consecutive patients undergoing TKA were divided into three groups, with 25 patients in each group receiving one of three implant designs: 25 Stryker Triathlon® CR (Stryker, Mahwah, NJ), 25 Zimmer NexGen® Flex CR (Zimmer, Warsaw, IN), and 25 StelKast Proven Gen-FlexTM CR (StelKast, Pittsburgh, PA). Flexion-extension gap matching was determined using only the medial flexion and extension gaps. Accuracy was determined by comparing actual resection thickness to desired resection thickness. "Optimal" accuracy was considered to be within 1.0mm of desired, and "near-optimal" accuracy was considered to be within 2.0mm of the desired resection thickness. Precision was determined by the variability of resection thicknesses within each system. RESULTS: Data demonstrated a lack of accuracy and precision across all three tested systems, with each system resulting in certain unique tendencies. Only one out of 75 cases resulted in optimal resection accuracy with all three cuts (Zimmer). When lowering the threshold to include both optimal and near-optimal (within 2 mm of error) with all three cuts, only one third of Stryker and Zimmer cases and two thirds of StelKast cases achieved this threshold, representing 44% of cases (33/75). CONCLUSIONS: Improvements in instrumentation to increase accuracy and precision may be warranted. Errors in fixation may be due to the instrumentation itself, and altering instrumentation to include less modularity, provide more stable fixation, and more reliably seal the implant on the femur may be of benefit.


Assuntos
Artroplastia do Joelho , Fêmur/cirurgia , Artroplastia do Joelho/instrumentação , Artroplastia do Joelho/métodos , Artroplastia do Joelho/normas , Artroplastia do Joelho/estatística & dados numéricos , Humanos , Articulação do Joelho/cirurgia , Prótese do Joelho
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