[The morbidity of the posterior iliac crest bone graft harvesting could be an overestimated fact. Study in patients with open posterolateral lumbar fusion]. / La morbilidad del sitio donante de injerto de cresta ilíaca posterior podría ser un hecho sobreestimado. Estudio en pacientes con fusión lumbar posterolateral abierta.

INTRODUCTION: spinal fusion is used to treat, among other pathologies, the degenerative intervertebral disc disease. Autologous iliac crest bone grafting is the golden standard treatment for increasing the rate of fusion; however, it isn't free of complications. OBJECTIVES: to investigate whether patients who have posterior iliac crest graft harvesting and are blinded to the donor site, can identify from which side the graft was harvested, and whether the intensity of this pain is related to the amount of graft obtained. MATERIAL AND METHODS: prospective, experimental, randomized and comparative, single-blind study. Adult patients who underwent primary instrumented open posterolateral lumbar fusion with autologous iliac crest bone graft between July 2019 and April 2020 were included. Patients were divided into two randomized groups. The amount of graft to be harvested was according to surgical needs. The patients were asked about pain according to the visual analogue scale in the first, third and sixth months after surgery, always requesting that the most painful side be identified. RESULTS: a total of 44 patients (n = 23 right crest, n = 21 left crest) were analyzed. Most patients were unable to identify the side from which the bone graft was harvested, with a statistically significant difference (p-value 0.0001). CONCLUSION: iliac crest bone graft harvesting is an effective and safe procedure that improves the rates of fusion without increasing the patient's morbidity.

INTRODUCCIÓN: la fusión espinal es utilizada para tratar, entre otras patologías, las enfermedades degenerativas discales. El injerto óseo autólogo de cresta ilíaca es el estándar de oro para aumentar las probabilidades de fusión; sin embargo, no está exento de complicaciones. OBJETIVOS: investigar si los pacientes a los que se les recolecta injerto de cresta ilíaca posterior y son cegados al lado del sitio donante, pueden identificar de qué lado se tomó dicho injerto y si la intensidad de este dolor está en relación con la cantidad de injerto obtenido. MATERIAL Y MÉTODOS: trabajo prospectivo, experimental, aleatorizado y comparativo, simple ciego. Se incluyeron pacientes adultos operados entre Julio de 2019 hasta Abril de 2020, a los cuales se les realizó una cirugía primaria de artrodesis lumbar posterolateral abierta instrumentada, con colocación de injerto óseo autólogo de cresta ilíaca. Se dividió a los pacientes en dos grupos aleatorizados. La cantidad de injerto a recolectar fue de acuerdo a las necesidades quirúrgicas. Se interrogó por el dolor según escala visual analógica al primer, tercer y sexto mes de la cirugía, siempre solicitando que se identifique el lado más doloroso. RESULTADOS: se analizaron 44 pacientes (n = 23 cresta derecha, n = 21 cresta izquierda). La mayoría de los pacientes no lograron identificar el lado del cual se obtuvo el injerto óseo, con una diferencia estadística ampliamente significativa (p = 0.0001). CONCLUSIÓN: la obtención de injerto óseo de cresta ilíaca es un procedimiento efectivo y seguro, que mejora las probabilidades de fusión sin aumentar la morbilidad del paciente.

[Is the Michigan State University lumbar disc herniation classification reliable among spine surgeons?] / ¿Es confiable la clasificación de la hernia de disco lumbar de la Universidad Estatal de Míchigan entre cirujanos de columna?

INTRODUCTION: there are numerous classifications for herniated discs, such classifications guide professionals about the severity of the lesion, the possible clinical picture of the patient, the most appropriate treatment and are undoubtedly a predictive tool to project the possible results of the therapies used. The main purpose of this work is to validate the inter- and intra-observer reliability of the Michigan State University (MSU) classification among the spine surgeons of our service and also to know the risk factors associated with the patients who underwent lumbar discectomy, the most affected disc level, the clinical presentation and the previous treatments performed in the patients treated by our team. MATERIAL AND METHODS: 50 nuclear magnetic resonance (MR) images in axial T2 section corresponding to the "maximum disc herniation" level were selected from patients diagnosed and submitted to meningo-radicular release surgery and single level lumbar discectomy retrospectively in the last two years from our database; these images were distributed among three spine surgeons of our institution. The three spine surgeons gave a specific classification for each MR image based on the MSU classification, then at an interval of seven days one of the three surgeons reclassified the images. The degree of agreement between surgeons was analyzed by calculating interobserver and intraobserver reliability using kappa statistical analysis. RESULTS: the analysis of the kappa coefficient indicated that most of the comparisons by observer gave a "good" concordance strength, the kappa index was higher than 0.64 in all the possible comparisons of the observations. In relation to the number of coincidences, in 60% of the patients there was a total coincidence between the three surgeons, with two coincidences in 24%, and in 16% there was no coincidence at all. For the intraobserver analysis the kappa index was 0.953 with a very good concordance strength, the observed agreement was 96%. CONCLUSION: our research shows a good reliability in the MSU classification among spine surgeons of our institution, as well as very good when reclassifying the intraobserver; we believe that having a sagittal MRI slice to classify them would be very useful, more research is needed to give a prognostic value to the location and size of the hernia and its relation with the surgical indication.

INTRODUCCIÓN: existen numerosas clasificaciones para las hernias de disco, dichas clasificaciones orientan a los profesionales acerca de la gravedad de la lesión, del posible cuadro clínico del paciente, del tratamiento más adecuado y constituyen, sin lugar a dudas, una herramienta predictiva para proyectar los posibles resultados de las terapias utilizadas. El propósito principal de este trabajo es validar la confiabilidad interobservador e intraobservador de la clasificación de la Universidad Estatal de Míchigan (MSU) entre los cirujanos de columna de nuestro servicio y además, conocer los factores de riesgo asociados con los pacientes que fueron intervenidos con discectomía lumbar, el nivel de disco más afectado, la presentación clínica y los tratamientos previos realizados en los pacientes tratados por nuestro equipo. MATERIAL Y MÉTODOS: fueron seleccionadas 50 imágenes de resonancia magnética nuclear (RM) en el corte axial de T2 correspondiente al nivel de "hernia discal máxima" de pacientes diagnosticados y sometidos a cirugía de liberación meningo radicular y discectomía lumbar de un solo nivel de manera retrospectiva en los últimos dos años de nuestra base de datos, estas imágenes se distribuyeron entre tres cirujanos especialistas en columna de nuestra institución. Los tres cirujanos de columna dieron una clasificación específica para cada imagen de RM basada en la clasificación de la MSU, luego en un intervalo de siete días uno de los tres cirujanos volvió a clasificar las imágenes. El grado de acuerdo entre los cirujanos se analizó calculando la confiabilidad interobservador e intraobservador mediante el análisis estadístico kappa. RESULTADOS: el análisis del coeficiente de kappa indicó que en la mayoría de las comparaciones por observador dieron una fuerza de concordancia "buena", el índice de kappa fue superior a 0.64 en todas las posibles comparaciones de las observaciones. En relación con la cantidad de coincidencias, en 60% de los pacientes hubo una coincidencia total entre los tres cirujanos, con dos coincidencias 24% y en 16% no hubo ninguna coincidencia. Para el análisis intraobservador el índice de kappa fue de 0.953 con una fuerza de concordancia muy buena, el acuerdo observado fue de 96%. CONCLUSIÓN: nuestra investigación demuestra una confiabilidad buena en la clasificación de la MSU entre cirujanos de columna de nuestra institución así como muy buena al reclasificar el intraobservador. Creemos que tener un corte sagital de RM para clasificarlas sería de gran utilidad. Se necesitan más investigaciones para dar un valor pronóstico a la ubicación y tamaño de la hernia y su relación con la indicación quirúrgica.

¿Es confiable la clasificación de la hernia de disco lumbar de la Universidad Estatal de Míchigan entre cirujanos de columna? / Is the Michigan State University lumbar disc herniation classification reliable among spine surgeons?

Resumen: Introducción: Existen numerosas clasificaciones para las hernias de disco, dichas clasificaciones orientan a los profesionales acerca de la gravedad de la lesión, del posible cuadro clínico del paciente, del tratamiento más adecuado y constituyen, sin lugar a dudas, una herramienta predictiva para proyectar los posibles resultados de las terapias utilizadas. El propósito principal de este trabajo es validar la confiabilidad interobservador e intraobservador de la clasificación de la Universidad Estatal de Míchigan (MSU) entre los cirujanos de columna de nuestro servicio y además, conocer los factores de riesgo asociados con los pacientes que fueron intervenidos con discectomía lumbar, el nivel de disco más afectado, la presentación clínica y los tratamientos previos realizados en los pacientes tratados por nuestro equipo. Material y métodos: Fueron seleccionadas 50 imágenes de resonancia magnética nuclear (RM) en el corte axial de T2 correspondiente al nivel de «hernia discal máxima¼ de pacientes diagnosticados y sometidos a cirugía de liberación meningo radicular y discectomía lumbar de un solo nivel de manera retrospectiva en los últimos dos años de nuestra base de datos, estas imágenes se distribuyeron entre tres cirujanos especialistas en columna de nuestra institución. Los tres cirujanos de columna dieron una clasificación específica para cada imagen de RM basada en la clasificación de la MSU, luego en un intervalo de siete días uno de los tres cirujanos volvió a clasificar las imágenes. El grado de acuerdo entre los cirujanos se analizó calculando la confiabilidad interobservador e intraobservador mediante el análisis estadístico kappa. Resultados: El análisis del coeficiente de kappa indicó que en la mayoría de las comparaciones por observador dieron una fuerza de concordancia «buena¼, el índice de kappa fue superior a 0.64 en todas las posibles comparaciones de las observaciones. En relación con la cantidad de coincidencias, en 60% de los pacientes hubo una coincidencia total entre los tres cirujanos, con dos coincidencias 24% y en 16% no hubo ninguna coincidencia. Para el análisis intraobservador el índice de kappa fue de 0.953 con una fuerza de concordancia muy buena, el acuerdo observado fue de 96%. Conclusión: Nuestra investigación demuestra una confiabilidad buena en la clasificación de la MSU entre cirujanos de columna de nuestra institución así como muy buena al reclasificar el intraobservador. Creemos que tener un corte sagital de RM para clasificarlas sería de gran utilidad. Se necesitan más investigaciones para dar un valor pronóstico a la ubicación y tamaño de la hernia y su relación con la indicación quirúrgica.

Abstract: Introduction: There are numerous classifications for herniated discs, such classifications guide professionals about the severity of the lesion, the possible clinical picture of the patient, the most appropriate treatment and are undoubtedly a predictive tool to project the possible results of the therapies used. The main purpose of this work is to validate the inter- and intra-observer reliability of the Michigan State University (MSU) classification among the spine surgeons of our service and also to know the risk factors associated with the patients who underwent lumbar discectomy, the most affected disc level, the clinical presentation and the previous treatments performed in the patients treated by our team. Material and methods: 50 nuclear magnetic resonance (MR) images in axial T2 section corresponding to the «maximum disc herniation¼ level were selected from patients diagnosed and submitted to meningo-radicular release surgery and single level lumbar discectomy retrospectively in the last two years from our database; these images were distributed among three spine surgeons of our institution. The three spine surgeons gave a specific classification for each MR image based on the MSU classification, then at an interval of seven days one of the three surgeons reclassified the images. The degree of agreement between surgeons was analyzed by calculating interobserver and intraobserver reliability using kappa statistical analysis. Results: The analysis of the kappa coefficient indicated that most of the comparisons by observer gave a «good¼ concordance strength, the kappa index was higher than 0.64 in all the possible comparisons of the observations. In relation to the number of coincidences, in 60% of the patients there was a total coincidence between the three surgeons, with two coincidences in 24%, and in 16% there was no coincidence at all. For the intraobserver analysis the kappa index was 0.953 with a very good concordance strength, the observed agreement was 96%. Conclusion: Our research shows a good reliability in the MSU classification among spine surgeons of our institution, as well as very good when reclassifying the intraobserver; we believe that having a sagittal MRI slice to classify them would be very useful, more research is needed to give a prognostic value to the location and size of the hernia and its relation with the surgical indication.

Traumatismo toracolumbar: fijación in situ vs descompresión y fijación en fracturas por estallido con compromiso neurológico / Thoracolumbar trauma: in situ fixation vs decompression and fixation in neurologically involved burst fractures

Resumen: Las fracturas por estallido toracolumbar son un gran desafío para los cirujanos de columna, es por ello que el objetivo de este estudio fue realizar una revisión sistemática analizando la literatura actual respecto a los diferentes factores a tener en cuenta en el manejo de las mismas. Material y métodos: Búsqueda sistemática en los sitios PubMed, Cochrane, MedlinePlus. «Thoracolumbar Burst Fractures¼ fueron las palabras claves. Se analizaron los títulos, resúmenes y texto completo a partir de dos observadores independientes. Resultados: Seis estudios han sido incluidos, cinco (83.33%) fueron de cohorte prospectivos y sólo uno (16.67%) retrospectivo. Hubo un total de 289 pacientes entre todas las muestras. La media de edad fue 38.48 años y la relación respecto al sexo fue masculino-femenino de 1.7:1. Conclusión: Las fracturas por estallido toracolumbar siguen siendo controversiales en su tratamiento con bibliografía publicada de bajo nivel de evidencia y sin establecer un consenso en el momento de la toma de decisiones con diferentes opciones según la experiencia y preferencia del cirujano. Sin embargo, la mayoría de los mismos se inclinan hacia el tratamiento con fijación posterior de segmento corto pero sin seguimiento a largo plazo. Respecto al déficit neurológico, la descompresión quirúrgica logró consenso entre todos los cirujanos.

Abstract: Thoracolumbar burst fractures are a great challenge for spine surgeons and that is why the aim of this study was to perform a systematic review analyzing the current literature regarding the different factors to take into account in their management. Material and methods: Systematic search in PubMed, Cochrane, MedlinePlus sites. «Thoracolumbar Burst Fractures¼ were the keywords. Titles, abstracts and full text were analyzed from two independent observers. Results: Six studies were included, five (83.33%) were prospective cohort studies and only one (16.67%) was retrospective. There were a total of 289 patients among all samples. The mean age was 38.48 years and the sex ratio was male to female of 1.7:1. Conclusion: Thoracolumbar burst fractures remain controversial in their treatment with published literature of low level of evidence and without establishing a consensus at the time of decision making with different options depending on the surgeon's experience and preference. However, most of them are inclined towards treatment with short segment posterior fixation but without long-term follow-up. Regarding neurological deficit, surgical decompression achieved consensus among all surgeons.

[Thoracolumbar trauma: in situ fixation vs decompression and fixation in neurologically involved burst fractures]. / Traumatismo toracolumbar: fijación in situ vs descompresión y fijación en fracturas por estallido con compromiso neurológico.

Thoracolumbar burst fractures are a great challenge for spine surgeons and that is why the aim of this study was to perform a systematic review analyzing the current literature regarding the different factors to take into account in their management. Material and methods: Systematic search in PubMed, Cochrane, MedlinePlus sites. "Thoracolumbar Burst Fractures" were the keywords. Titles, abstracts and full text were analyzed from two independent observers. Results: Six studies were included, five (83.33%) were prospective cohort studies and only one (16.67%) was retrospective. There were a total of 289 patients among all samples. The mean age was 38.48 years and the sex ratio was male to female of 1.7:1. Conclusion: Thoracolumbar burst fractures remain controversial in their treatment with published literature of low level of evidence and without establishing a consensus at the time of decision making with different options depending on the surgeon's experience and preference. However, most of them are inclined towards treatment with short segment posterior fixation but without long-term follow-up. Regarding neurological deficit, surgical decompression achieved consensus among all surgeons.

Las fracturas por estallido toracolumbar son un gran desafío para los cirujanos de columna, es por ello que el objetivo de este estudio fue realizar una revisión sistemática analizando la literatura actual respecto a los diferentes factores a tener en cuenta en el manejo de las mismas. Material y métodos: Búsqueda sistemática en los sitios PubMed, Cochrane, MedlinePlus. "Thoracolumbar Burst Fractures" fueron las palabras claves. Se analizaron los títulos, resúmenes y texto completo a partir de dos observadores independientes. Resultados: Seis estudios han sido incluidos, cinco (83.33%) fueron de cohorte prospectivos y sólo uno (16.67%) retrospectivo. Hubo un total de 289 pacientes entre todas las muestras. La media de edad fue 38.48 años y la relación respecto al sexo fue masculino-femenino de 1.7:1. Conclusión: Las fracturas por estallido toracolumbar siguen siendo controversiales en su tratamiento con bibliografía publicada de bajo nivel de evidencia y sin establecer un consenso en el momento de la toma de decisiones con diferentes opciones según la experiencia y preferencia del cirujano. Sin embargo, la mayoría de los mismos se inclinan hacia el tratamiento con fijación posterior de segmento corto pero sin seguimiento a largo plazo. Respecto al déficit neurológico, la descompresión quirúrgica logró consenso entre todos los cirujanos.

[Application of vancomycin powder on graft. Does the fusion of the rabbits spine is affected?] / Aplicación de vancomicina en polvo sobre el injerto. ¿Afecta la fusión de columna en conejos?

INTRODUCTION: In vitro studies suggest that vancomycin is highly cytotoxic for osteoblasts. However, several clinical studies in vivo have not provided evidence or determined that effect on osteogenesis. The aim of this work was to investigate the effect of topical vancomycin on a lumbar arthrodesis model on New Zealand white rabbits. MATERIAL AND METHODS: Double-blind randomized experimental study. Posterolateral spinal fusion was performed on 30 rabbits, divided into two groups, A: graft/placebo, B: graft/vancomycin. After sacrificing them, the bone callus was evaluated with axial tomography and classified into three groups: no fusion (0), partial/incomplete fusion (1) and fusion (2). The samples were also histologically analyzed. The associations between the presence of fusion (complete/incomplete) and the group tested were estimated using Poisson log-linear models with two covariates. Adherences to histologically obtained responses were studied using contingency tables and 2 tests. The significance level was set equal to 0.05. RESULTS: Treatment with vancomycin has 30% (0.30. CI 95%: 0.12-0.94) less chance compared to the placebo group, of presenting complete fusion. In other words, the vancomycin group has 2.3 times (CI 95%: 1.02-4.91) more likelyhood, compared to placebo, to have incomplete fusion. CONCLUSION: The application of vancomycin powder mixed with graft reduces fusion rates by 30%, but at doses 5 times higher than those routinely used in arthrodesis by the authors medical team.

INTRODUCCIÓN: Estudios in vitro sugieren que la vancomicina es altamente citotóxica para los osteoblastos. No obstante, diversos estudios clínicos in vivo no han aportado evidencias o determinado ese efecto en la osteogénesis. El objetivo del presente trabajo fue investigar el efecto de la vancomicina tópica, en un modelo de artrodesis lumbar sobre conejos blancos neozelandeses. MATERIAL Y MÉTODOS: Estudio experimental randomizado doble ciego. Se realizó fusión espinal posterolateral a 30 conejos, divididos en dos grupos, A: injerto/placebo, B: injerto/vancomicina. Luego de sacrificarlos, se evaluó el callo óseo con tomografía axial, clasificándolos en tres grupos: sin fusión (0), fusión parcial/incompleta (1) y fusión (2). Las muestras también fueron analizadas histológicamente. Las asociaciones entre presencia de fusión (completa/incompleta) y el grupo ensayado fueron estimadas con modelos log-lineales de Poisson con dos covariables. Las adherencias con las respuestas obtenidas histológicamente fueron estudiadas usando tablas de contingencia y test 2. El nivel de significación se fijó igual a 0.05. RESULTADOS: El tratamiento con vancomicina tiene 30% (0.30. IC 95%: 0.12-0.94) menos probabilidad respecto al grupo placebo de presentar fusión completa. En otros términos, el grupo vancomicina tiene 2.3 veces (IC 95%: 1.02-4.91) más probabilidad, respecto del placebo, de tener fusión incompleta. CONCLUSIÓN: La aplicación de vancomicina en polvo mezclada con injerto reduce 30% las tasas de fusión, pero a dosis cinco veces más alta que las usadas rutinariamente en artrodesis por el equipo médico de los autores.

Periprostatic adipose inflammation is associated with high-grade prostate cancer.

BACKGROUND: Obesity, a cause of subclinical inflammation, is associated with increased risk of high-grade prostate cancer (PC) and poor outcomes. Whether inflammation occurs in periprostatic white adipose tissue (WAT), and contributes to the negative impact of obesity on PC aggressiveness, is unknown. METHODS: In a single-center, cross-sectional design, men with newly diagnosed PC undergoing radical prostatectomy were eligible for study participation. The primary objective was to examine the prevalence of periprostatic WAT inflammation defined by the presence of crown-like structures (CLS-P) as detected by CD68 immunohistochemistry. Secondary objectives were to explore the clinical and systemic correlates of periprostatic WAT inflammation. Tumor characteristics and host factors including BMI, adipocyte diameter, and circulating levels of lipids, adipokines, and other metabolic factors were measured. Wilcoxon rank-sum, Chi-square, or Fisher's exact tests, and generalized linear regression were used to examine the association between WAT inflammation and tumor and host characteristics. RESULTS: Periprostatic fat was collected from 169 men (median age 62 years; median BMI 28.3). Periprostatic WAT inflammation was identified in 49.7% of patients and associated with higher BMI (P=0.02), larger adipocyte size (P=0.004) and Gleason grade groups IV/V tumors (P=0.02). The relationship between WAT inflammation and high Gleason grade remained significant after adjusting for BMI (P=0.04). WAT inflammation correlated with higher circulating levels of insulin, triglycerides, and leptin/adiponectin ratio, and lower high density lipoprotein cholesterol, compared to those without WAT inflammation (P's <0.05). CONCLUSION: Periprostatic WAT inflammation is common in this cohort of men with PC and is associated with high-grade PC.

Risk factors associated with sensitive skin and potential role of lifestyle habits: a cross-sectional study.

Sensitive skin (SS) is a widespread condition, but still not completely understood. To identify risk factors that increase the likelihood of SS, 258 women aged between 20 and 65 years old and resident in the Netherlands were surveyed by questionnaire, which included questions on sociodemographic characteristics (age group, Fitzpatrick skin type, hormonal status), health state (atopic predisposition, skin diseases) and lifestyle habits (history of smoking and of sun exposure, frequency of physical exercise). Analysis of the responses confirmed that atopic predisposition, presence of skin diseases and Fitzpatrick skin types I and II are risk factors significantly associated with SS. In addition, as current or past smoking and a history of low sun exposure showed a trend to increase the likelihood of reporting SS, we suggest that the potential role of lifestyle factors in the onset or exacerbation of SS should be investigated further.

Measurement of skin surface biomakers by Transdermal Analyses Patch following different in vivo models of irritation: a pilot study.

BACKGROUND/PURPOSE: FibroTx Transdermal Analyses Patch (TAP) is a novel technology for non-invasive measurements of protein biomarkers on the skin surface, in vivo. The aim of this study was to explore the potential of TAP in detecting skin surface biomarkers following mild perturbations, in vivo, using two experimental models: tape stripping, mimicking acute barrier disruption, and histamine iontophoresis, mimicking acute and local inflammation at minimal skin barrier insult. METHODS: Tape stripping and histamine iontophoresis were performed in two separate experiments on the volar forearm of healthy volunteers (n = 27 and n = 10, respectively). Biomarker levels were assessed with TAP at baseline and up to 72 h after stimulation. Functional (transepidermal water loss -TEWL- and a* value) and morphological (confocal reflectance microscopy -RCM) assessments were added in the tape stripping and histamine iontophoresis experiments, respectively. RESULTS: Cytokines IL-1α and IL-1RA and the antimicrobial peptide hBD-1 showed distinct dynamics, despite substantial inter-individual variation in levels, with an increase following tape stripping and a decrease following histamine iontophoresis. These dynamics could be related to the assessments made by TEWL and RCM. In the tape stripping experiment, additional biomarkers could be detected. CONCLUSION: TAP measurements, especially IL-1α, IL-1RA, and hBD-1, from the skin surface were sensitive enough for monitoring dynamic changes in the skin in the two models of skin perturbation. We conclude that TAP holds promise for non-invasively unraveling the dynamics of processes related to skin perturbation and repair.

The effects of nonsteroidal anti-inflammatory drugs on clinical outcomes, synovial fluid cytokine concentration and signal transduction pathways in knee osteoarthritis. A randomized open label trial.

OBJECTIVE: We investigated the effects of celecoxib, diclofenac, and ibuprofen on the disease-specific quality of life, synovial fluid cytokines and signal transduction pathways in symptomatic knee osteoarthritis (OA). DESIGN: Ninety patients scheduled for a total knee arthroplasty (TKA) were randomized to six groups that were treated with low and high dosages of celecoxib, diclofenac or ibuprofen. At the time of the first admission (T0) and at surgery (T1 = 14 days after beginning of the nonsteroidal anti-inflammatory drugs (NSAIDs)), samples of knee synovial fluid were obtained from each patient for analysis. During the surgery the synovial tissue was harvested from the knee of patients. The Western Ontario and McMaster universities (WOMAC) score was used to evaluate the patient disease-specific quality of life at T0 and T1. Microarray tests performed at T0 and T1 were used to evaluate the effects of NSAIDs on Tumor necrosis factor (TNF)-alpha, Interleukin-6 (IL-6), IL8 and Vascular endothelial growth factor (VEGF) concentration in the synovial fluid. Western blot assays evaluated the effects of NSAIDs on MAP kinase (MAPK) signal transduction pathway in the synovial membrane. RESULTS: NSAID treatment induced a statistically significant improvement in the WOMAC score and a statistically significant decrease in the IL-6, VEGF and TNF-alpha concentration in the synovial fluid. Higher dosages of NSAIDs provided a greater improvement in the disease-specific quality of life of patients and lower concentrations of pro-inflammatory cytokines in the synovial fluid. Inhibition of MAPKs was noted after NSAID treatment. CONCLUSION: Short-term NSAID treatment improves the patient disease-specific quality of life with a parallel decrease in pro-inflammatory synovial fluid cytokine levels in knee OA. Signal transduction pathways may be involved in regulating the anti-inflammatory effects of NSAIDs. ClinicalTrial.gov: NCT01860833.

Effects of simvastatin and rosuvastatin on RAS protein, matrix metalloproteinases and NF-κB in lung cancer and in normal pulmonary tissues.

OBJECTIVES: In this study, we have evaluated effects of 24-hour treatments with simvastatin or rosuvastatin on RAS protein, NF-κB and MMP expression in LC tissues obtained from 12 patients undergoing thoracic surgery. MATERIALS AND METHODS: Normal and lung tumour tissues obtained from each sample were exposed to simvastatin (2.5-30 µm) or rosuvastatin (1.25-30 µm) and western blot analysis was then performed. RESULTS: We documented increased expression of proteins, MMP-2, MMP-9 and NF-κB-p65 in LC tissues, with respect to normal tissues (P < 0.01). In the malignant tissues, simvastatin and rosuvastatin significantly (P < 0.01) and dose-dependently reduced RAS protein, MMP-2/9 and NF-κB-p65 expression. CONCLUSIONS: In conclusion, our results suggest that simvastatin and rosuvastatin could play a role in LC treatment by modulation of RAS protein, MMP-2/9 and NF-κB-p65.

Effects of statins and farnesyl transferase inhibitors on ERK phosphorylation, apoptosis and cell viability in non-small lung cancer cells.

OBJECTIVE: 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins) can affect post-translational processes, thus being responsible for decreased farnesylation and geranylgeranylation of intracellular small G proteins such as Ras, Rho and Rac, essential for cell survival and proliferation. In this regard, recent in vitro and in vivo studies suggest a possible role for both statins and farnesyl transferase inhibitors in the treatment of malignancies. Within such a context, the aim of our study was to investigate effects of either simvastatin (at concentrations of 1, 15, and 30 µm) or the farnesyl transferase inhibitor R115777 (at concentrations of 0.1, 1, and 10 µm), on two cultures of human non-small lung cancer cells, adenocarcinoma (GLC-82) and squamous (CALU-1) cell lines. In particular, we evaluated actions of these two drugs on phosphorylation of the ERK1/2 group of mitogen-activated protein kinases and on apoptosis, plus on cell numbers and morphology. MATERIALS AND METHODS: Western blotting was used to detect ERK phosphorylation, and to assess apoptosis by evaluating caspase-3 activation; apoptosis was also further assessed by terminal deoxynucleotidyl-mediated dUTP nick end labelling (TUNEL) assay. Cell counting was performed after trypan blue staining. RESULTS AND CONCLUSION: In both GLC-82 and CALU-1 cell lines, simvastatin and R115777 significantly reduced ERK phosphorylation; this effect, which reached the greatest intensity after 36 h treatment, was paralleled by a concomitant induction of apoptosis, documented by significant increase in both caspase-3 activation and TUNEL-positive cells, associated with a reduction in cell numbers. Our results thus suggest that simvastatin and R115777 may exert, in susceptible lung cancer cell phenotypes, a pro-apoptotic and anti-proliferative activity, which appears to be mediated by inhibition of the Ras/Raf/MEK/ERK signalling cascade.

Characteristics and clinical implications of the pharmacokinetic profile of ibuprofen in patients with knee osteoarthritis.

BACKGROUND AND OBJECTIVE: Ibuprofen is a non-selective cyclo-oxygenase (COX)-1/COX-2 inhibitor used to treat pain conditions and inflammation. Limited data have been published concerning the pharmacokinetic profile and clinical effects of ibuprofen in patients with osteoarthritis (OA). In this paper we compared the pharmacokinetic and clinical profile of ibuprofen (at a dosage of from 800 mg/day to 1800 mg/day) administered in patients affected by severe knee OA. METHODS: Ibuprofen was administered for 7 days to patients who were scheduled to undergo knee arthroplasty due to OA. After 7 days, the ibuprofen concentration in plasma and synovial fluid was measured through both high-performance liquid chromatography (HPLC)-UV and gas chromatography-mass spectroscopy (GC/MS), while clinical effects were evaluated through both visual analogue scale (VAS) and Western Ontario and McMaster Universities (WOMAC) scores. The Naranjo scale and the WHO causality assessment scale were used for estimating the probability of adverse drug reactions (ADRs). The severity of ADRs was assessed by the modified Hartwig and Siegel scale. RESULTS: Ibuprofen showed a dose-dependent diffusion in both plasma and synovial fluid, which was related to the reduction of pain intensity and improvement of health status, without the development of ADRs. CONCLUSION: Ibuprofen at higher dosages can be expected to provide better control of OA symptoms as a result of higher tissue distribution.

Natural Products Genomics: A novel approach for the discovery of anti-cancer therapeutics.

Plants continue to retain some advantages over combinatorial chemistry as sources of novel compounds, for example, they can generate metabolites with a complexity beyond synthetic chemistry. However, this comes with its own problems in production and synthetic modification of these compounds. Natural Products Genomics (NPG) aims to access the plants own genomic capacity to increase yields, and modify complex bioactive metabolites, to alleviate these limitations. NPG uses a combination of gain of function mutagenesis and selection to a) mimic the evolution of novel compounds in plants, and b) to increase yields of known bioactive metabolites. This process is performed rapidly at the cell culture level in large populations of mutants. Two examples demonstrating proof of concept in Nicotiana tabacum (tobacco) and proof of application in the medicinal plant species Catharanthus roseus, are included to illustrate the feasibility of this approach. This biotechnology platform may alter the way in which plant drug discovery is perceived by the pharmaceutical industry, and provides an alternative to combinatorial chemistry for the discovery, modification and production of highly complex bioactive molecules.

Effects of budesonide on P38 MAPK activation, apoptosis and IL-8 secretion, induced by TNF-alpha and Haemophilus influenzae in human bronchial epithelial cells.

Non-typeable Haemophilus influenzae (NTHi) is one of the most frequently involved pathogens in bacterial exacerbations of chronic obstructive pulmonary disease (COPD). In the airways, the main tissue target of NTHi is bronchial epithelium, where this pathogen can further amplify the inflammatory and structural changes induced by proinflammatory cytokines such as tumour necrosis factor-alpha (TNF-alpha). Therefore, the aim of this study is to investigate, in primary cultures of human bronchial epithelial cells, the effects of NTHi on signal transduction pathways, apoptotic events and chemokine production activated by TNF-alpha. Moreover, we also evaluated the effects exerted on such cellular and molecular phenomena by a corticosteroid drug. p38 mitogen-activated protein kinase (MAPK) phosphorylation was analyzed by Western blotting, using an anti-phospho-p38 MAPK monoclonal antibody. Apoptosis was assayed by active caspase-3 expression. Interleukin-8 (IL-8/CXCL8) was detected in cell-free culture supernatants by ELISA. TNF-alpha induced a significant increase in p38 MAPK phosphorylation. NTHi was able to potentiate the stimulatory actions of TNF-alpha on caspase-3 expression and, to a lesser extent, on IL-8 secretion. These effects were significantly (P less than 0.01) inhibited by a pharmacological pre-treatment with budesonide. These results suggest that TNF-alpha is able to stimulate, via activation of p38 MAPK signalling pathway, IL-8 release and airway epithelial cell apoptosis; the latter effect can be markedly potentiated by NTHi. Furthermore, budesonide can be very effective in preventing, through inhibition of p38 MAPK phosphorylation, both structural and proinflammatory changes elicited in bronchial epithelium by TNF-alpha and NTHi.

Interleukin-6 receptor superantagonist Sant7 inhibits TGF-beta-induced proliferation of human lung fibroblasts.

OBJECTIVES: Both interleukin-6 (IL-6) and transforming growth factor-beta (TGF-beta) are crucially involved in fibrotic events that characterize interstitial lung diseases (ILD). Therefore, the aim of this study was to investigate in primary cultures of normal and fibrotic human lung fibroblasts (HLF), exposed to either IL-6 or TGF-beta1, the effects on phosphorylation of mitogen-activated protein kinases (MAPK) and cell growth of IL-6 signalling inhibition, performed by the IL-6 receptor superantagonist Sant7. MATERIALS AND METHODS: MAPK phosphorylation was detected by Western blotting, HLF viability and proliferation were evaluated using the trypan blue staining and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, respectively. RESULTS: Sant7, at a concentration of 1 microg/mL, was capable of significantly inhibiting HLF proliferation and MAPK phosphorylation induced by cell exposure to IL-6 (100 ng/mL) or TGF-beta1 (10 ng/mL), whose actions were more evident in fibrotic cells. CONCLUSIONS: These findings suggest that, in HLFs derived from patients with ILDs, the proliferative mechanisms activated by TGF-beta1 are at least in part mediated by an increased release of IL-6, leading to phosphorylation-dependent MAPK activation. Such preliminary findings may thus open new therapeutic perspectives for fibrogenic ILDs, based on inhibition of signal transduction pathways stimulated by the IL-6 receptor.

Detection and prediction of alternative splicing in Arabidopsis thaliana.

Alternative splicing is an important process for increasing the diversity arising from a single gene. Presently, most studies aimed at detecting alternatively spliced genes use Expressed Sequence Tags (ESTs). However, the EST studies based on spliced transcripts analyse sequences by alignment rather than sequence patterns. Second, EST libraries can be of uncertain quality. To address these issues and to improve the quality of detection and prediction for alternative splicing, we propose a method that primarily uses pre-mRNAs. It is achieved by a decision tree algorithm using triplet nucleotides as attributes for each chromosome in Arabidopsis thaliana. In addition, we propose a novel algorithm for accurate prediction.