Dynamics of CD44+ bovine nucleus pulposus cells with inflammation.

Intervertebral Disc (IVD) degeneration has been associated with a chronic inflammatory response, but knowledge on the contribution of distinct IVD cells, namely CD44, to the progression of IVD degeneration remains elusive. Here, bovine nucleus pulposus (NP) CD44 cells were sorted and compared by gene expression and proteomics with the negative counterpart. NP cells were then stimulated with IL-1b (10 ng/ml) and dynamics of CD44 gene and protein expression was analyzed upon pro-inflammatory treatment. The results emphasize that CD44 has a multidimensional functional role in IVD metabolism, ECM synthesis and production of neuropermissive factors. CD44 widespread expression in NP was partially associated with CD14 and CD45, resulting in the identification of distinct cell subsets. In conclusion, this study points out CD44 and CD44-based cell subsets as relevant targets in the modulation of the IVD pro-inflammatory/degenerative cascade.

Can Specular Gloss Measurements Predict the Effectiveness of Finishing/Polishing Protocols in Dental Polymers? A Systematic Review and Linear Mixed-effects Prediction Model.

PURPOSE: The current gold standard measure to assess polishing efficacy is surface roughness (SR) assessed in laboratory research. Specular gloss (SG) has been negatively correlated to SR, which raises the following question: Can SG be used to accurately determine the effectiveness of a finishing/polishing procedure in direct resin composites? METHODS: A systematic approach and search strategy, following Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines, was developed and conducted in five electronic databases: PubMed/Medline, Scopus, Web of Science, EMBASE (Ovid), and SciELO/LILACS to identify laboratory studies that assessed SR and SG, simultaneously, of resin composites, without date or language restriction. Risk of bias assessment was carried out by two reviewers, independently. From the extracted quantitative data of SG/SR, regression analyses were performed, and a linear mixed-effects prediction model was derived using the nimble package in R (v4.0.3). RESULTS: A total of 928 potential studies were found, out of which, 13 were eligible after criterion screening. Experimental groups featured 31 resin composites of six different filler types, with the most common being microhybrids followed by nanohybrids. More than half of the studies initially reported a linear correlation between SR and SG, which ranged from r2 = 0.34-0.96. Taking into account the regression analysis and prediction model posteriorly performed, the corresponding SG threshold for 0.2 µm is estimated to be >55 GU. Most of the evidence was classified as moderate or high risk of bias. CONCLUSION: SG is universally correlated to SR in polymers, and a reference value of >55 GU is proposed, above which samples are considered well polished.

COPD phenotypes by computed tomography and ventilatory response to exercise.

INTRODUCTION: Computed tomographic (CT) phenotypic patterns of chronic obstructive pulmonary disease (COPD) identify different clinical features of disease. The impact of these variables on the physiological response to exercise has been the focus of a great deal of research as it allows more individualized clinical approaches. The aim of our study was to evaluate the relationships between CT phenotyping of subjects with COPD and the ventilatory response during cardiopulmonary exercise testing (CPET). METHODS: Subjects with COPD were classified into four phenotypes based on CT metrics of emphysema (low attenuation area less than a threshold of -950 Hounsfield [%LAA-950]) and airwall thickness (bronchial wall area percentage [%WA]). RESULTS: Eighty COPD patients (78.8% males, median age 65±11.3 years) were enrolled in the study. Based on CT phenotype, 25 (31.3%) patients were classified as normal, 27 (33.8%) air dominant, 17 (21.3%) emphysema dominant and 11 (13.8%) mixed type. The emphysema and mixed phenotypes showed the highest ventilatory equivalent for carbon dioxide (VE/VCO2) and VE/VCO2 slope (p<0,05). In all phenotypes, %LAA was positive correlated with VE/VCO2 and VE/VCO2 slope (r = 0.437, p = 0.006 and r = 0.503, p<0.001, respectively). %WA also showed a positive correlation with VE/VCO2 and VE/VCO2 slope (r = 0.541, p<0.001 and r = 0.299, p = 0.033, respectively). In multivariate regression models, after adjustment for age, BMI, sex and FEV1, %LAA was the only independent predictor of VE/VCO2 and VE/VCO2 slope (ß 0.343, SE 0.147, 95% CI 0.009/0.610, p = 0.044 and ß 0.496, SE 0.081, 95% CI 0.130/0.455, p = 0.001, respectively). CONCLUSION: Emphysema (%LAA) and airways metrics (%WA) had strong relationships with the different characteristics of ventilatory response to exercise in subjects with mild to moderate COPD. In particular, %LAA seemed to play an important role as an independent predictor of VE/VCO2 and VE/VCO2 slope. These results suggested that CT phenotyping may help predicting ventilatory response to exercise in subjects with COPD.

Intervertebral disc decellularisation: progress and challenges.

Intervertebral disc (IVD) degeneration and the consequent low-back pain (LBP) affect over 80 % of people in western societies, constituting a tremendous socio-economic burden worldwide and largely impairing patients' life quality. Extracellular matrix (ECM)-based scaffolds, derived from decellularised tissues, are being increasingly explored in regenerative medicine for tissue repair. Decellularisation plays an essential role for host cells and antigen removal, while maintaining native microenvironmental signals, including ECM structure, composition and mechanical properties, which are essential for driving tissue regeneration. With the lack of clinical solutions for IVD repair/regeneration, implantation of decellularised IVD tissues has been explored to halt and/or revert the degenerative cascade and the associated LBP symptoms. Over the last few years, several researchers have focused on the optimisation of IVD decellularisation methods, combining physical, chemical and enzymatic treatments, in order to successfully develop a cell-free matrix. Recellularisation of IVD-based scaffolds with different cell types has been attempted and numerous methods have been explored to address proper IVD regeneration. Herein, the advances in IVD decellularisation methods, sterilisation procedures, repopulation and biocompatibility tests are reviewed. Additionally, the importance of the donor profile for therapeutic success is also addressed. Finally, the perspectives and major hurdles for clinical use of the decellularised ECM-based biomaterials for IVD are discussed. The studies reviewed support the notion that tissue-engineering-based strategies resorting to decellularised IVD may represent a major advancement in the treatment of disc degeneration and consequent LBP.

IL-1ß-pre-conditioned mesenchymal stem/stromal cells' secretome modulates the inflammatory response and aggrecan deposition in intervertebral disc.

Mesenchymal stem/stromal cells (MSCs) have been increasingly used in clinical trials for low-back pain (LBP) and intervertebral disc (IVD) degeneration with promising results. Their action mechanisms are not fully understood, but they reduce IVD pro-inflammatory markers in a pro-inflammatory/degenerative IVD microenvironment. In this study the therapeutic potential of the MSC secretome, as an alternative cell-free approach for treating degenerated IVDs, was examined. Human bone marrow-derived MSC secretome (MSCsec) was collected after 48 h of preconditioning in IL-1ß (10 ng/mL) and low oxygen (6 % O2), mimicking the degenerative IVD. IL-1ß-pre-conditioning of MSCs increased secretion of pro-inflammatory markers hIL-6, hIL-8, hMCP-1, etc. The therapeutic effect of MSCsec was tested in a pro-inflammatory/degenerative IVD ex vivo model. MSCsec down-regulated IVD gene expression of pro-inflammatory cytokines (bIL-6, bIL-8) and matrix degrading enzyme bMMP1, while bMMP3 and bTIMP2 were up-regulated, at 48 h. After 14 d, MSCsec-treated IVDs revealed increased aggrecan deposition, although no differences in other ECM components were observed. Protein analysis of the MSCsec-treated IVD supernatant revealed a significant increase of CXCL1, MCP-1, MIP-3α, IL-6, IL-8 and GRO α/ß/γ (related to TNF, NOD-like receptor and neutrophil chemotaxis signalling), and a decrease of IFN-γ, IL-10, IL-4, IL-5 and TNF-α (associated with T-cell receptor signalling). MSCsec-treated IVD supernatants did not promote angiogenesis and neurogenesis in vitro. Overall, MSCsec can be a safe therapeutic approach, presenting a strong immunomodulatory role in degenerated IVD while potentiating aggrecan deposition, which can open new perspectives on the use of MSCsec as a cell-based/ cell-free therapeutic approach to LBP.

Three-dimensional scaffolds of fetal decellularized hearts exhibit enhanced potential to support cardiac cells in comparison to the adult.

A main challenge in cardiac tissue engineering is the limited data on microenvironmental cues that sustain survival, proliferation and functional proficiency of cardiac cells. The aim of our study was to evaluate the potential of fetal (E18) and adult myocardial extracellular matrix (ECM) to support cardiac cells. Acellular three-dimensional (3D) bioscaffolds were obtained by parallel decellularization of fetal- and adult-heart explants thereby ensuring reliable comparison. Acellular scaffolds retained main constituents of the cardiac ECM including distinctive biochemical and structural meshwork features of the native equivalents. In vitro, fetal and adult ECM-matrices supported 3D culture of heart-derived Sca-1(+) progenitors and of neonatal cardiomyocytes, which migrated toward the center of the scaffold and displayed elongated morphology and excellent viability. At the culture end-point, more Sca-1(+) cells and cardiomyocytes were found adhered and inside fetal bioscaffolds, compared to the adult. Higher repopulation yields of Sca-1(+) cells on fetal ECM relied on ß1-integrin independent mitogenic signals. Sca-1(+) cells on fetal bioscaffolds showed a gene expression profile that anticipates the synthesis of a permissive microenvironment for cardiomyogenesis. Our findings demonstrate the superior potential of the 3D fetal microenvironment to support and instruct cardiac cells. This knowledge should be integrated in the design of next-generation biomimetic materials for heart repair.

ATM down-regulation is associated with poor prognosis in sporadic breast carcinomas.

BACKGROUND: Ataxia telangiectasia-mutated (ATM) gene downexpression has been reported in sporadic breast carcinomas (BC); however, the prognostic value and mechanisms of ATM deregulation remain unclear. PATIENTS AND METHODS: ATM and miRNAs (miR-26a, miR-26b, miR-203, miR-421, miR-664, miR-576-5p and miR-18a) expression levels were evaluated by quantitative real-time PCR (RT-qPCR) in 52 BC and 3 normal breast samples. ATM protein expression was assessed by immunohistochemistry in 968 BC and 35 adjacent normal breast tissues. ATM copy number alteration was detected by array comparative genomic hybridization (aCGH) in 42 tumours. RESULTS: Low ATM levels were associated with tumour grade. Absence of ATM protein expression was associated with distant metastasis (P < 0.001), reduced disease-free survival (DFS, P < 0.001) and cancer-specific survival (CSS, P < 0.001). Multivariate analysis indicated ATM protein expression as an independent prognostic marker for DFS (P = 0.001, HR = 0.579) and CSS (P = 0.001, HR = 0.554). ATM copy number loss was detected in 12% of tumours and associated with lower mRNA levels. miR-421 over-expression was detected in 36.5% of cases which exhibit lower ATM transcript levels (P = 0.075, r = -0.249). CONCLUSIONS: The data suggest that ATM protein expression is an independent prognostic marker in sporadic BC. Gene copy number loss and miR-421 over-expression may be involved in ATM deregulation in BC.

Postprandial insulin resistance in Zucker diabetic fatty rats is associated with parasympathetic-nitric oxide axis deficiencies.

The Zucker diabetic fatty (ZDF) rat is an obesity and type 2 diabetes model. Progression to diabetes is well characterised in ZDF rats, but only in the fasted state. We evaluated the mechanisms underlying postprandial insulin resistance in young ZDF rats. We tested the hypothesis that the overall postprandial action of insulin is affected in ZDF rats as a result of impairment of the hepatic parasympathetic-nitric oxide (PSN-NO) axis and/or glutathione (GSH), resulting in decreased indirect (PSN-NO axis) and direct actions of insulin. Nine-week-old male ZDF rats and lean Zucker rats (LZR, controls) were used. The action of insulin was assessed in the fed state before and after parasympathetic antagonism atropine. Basal hepatic NO and GSH were measured, as well as NO synthase (NOS) and γ-glutamyl-cysteine synthethase (GCS) activity and expression. ZDF rats presented postprandial hyperglycaemia (ZDF, 201.4 ± 12.9 mg/dl; LZR, 107.7 ± 4.3 mg/dl), but not insulinopaenia (ZDF, 5.9 ± 0.8 ng/ml; LZR, 1.5 ± 0.3 ng/ml). Total postprandial insulin resistance was observed (ZDF, 78.6 ± 7.5 mg glucose/kg; LZR, 289.2 ± 24.7 mg glucose/kg), with a decrease in both the direct action of insulin (ZDF, 54.8 ± 7.0 mg glucose/kg; LZR, 173.3 ± 20.5 mg glucose/kg) and the PSN-NO axis (ZDF, 24.5 ± 3.9 mg glucose/kg; LZR, 115.9 ± 19.4 mg glucose/kg). Hepatic NO (ZDF, 117.2 ± 11.4 µmol/g tissue; LZR, 164.6 ± 4.9 µmol/g tissue) and GSH (ZDF, 4.9 ± 0.3 µmol/g; LZR, 5.9 ± 0.2 µmol/g) were also compromised as a result of decreased NOS and GCS activity, respectively. These results suggest a compromise of the mechanism responsible for potentiating insulin action after a meal in ZDF rats. We show that defective PSN-NO axis and GSH synthesis, together with an impaired direct action of insulin, appears to contribute to postprandial insulin resistance in this model.

[Minimally invasive thoracic surgery: a fifteen-years experience]. / Cirurgia torácica minimamente invasiva. Experiêcia de 15 anos.

The evolution of surgical techniques has rendering possible for thoracic surgery to be performed efficiently and safely, with less aggressive approaches. Being less traumatic, minimally invasive surgery allows a faster post-operative recovery, less complications and reduces in patient days and morbidity. The authors consider as minimally invasive approaches the video-assisted thoracic surgery, as well as the minithoractomy and the ministernotomy. The present paper reviews thoracic surgeries performed for the last 15 years in a cardiothoracic surgical unit using a minimally invasive surgical approach.

[Intra-thoracic goiter: analysis of the Department's experience]. / Bócio mergulhante: Análise da casuística do Serviço.

A review of all clinical files with pre-operative diagnosis of intra-thoracic goiter operated in our Department from 2001-2009 was made. We identified 30 patients, of which 66% were females. 63% of the patients were asymptomatic, 46% had clinical or radiological findings of mediastinal structures compression and 2 patients were ventilated prior to surgery. 50% of the patients were operated on through an inferior transverse cervicotomy, the remaining were operated on through partial or complete sternotomy, with cervical approach. 6,6% of patients (n=2) had an intra-operative diagnosis of invasive thymoma, 3,3% (n=1) had a very invasive mediastinal tumor and 3,3% (n=1) had an ectopic thyroid. Average surgical length of time was of 83 minutes. In 10% of patients (n=3) post-operative invasive ventilation was required, one of which for unilateral permanent lesion of the recurrent laryngeal nerve. In the long term follow-up we assessed 18 patients. The median follow up was 53 months. One patient died at the 15th post-operatory day of pneumonia, one expired at the 30th day for anaplasic thyroid carcinoma progression and one died at the 18 month from invasive thymoma progression. One patient currently reveals evidence of local recurrence 3 patients are currently taking levothyroxine and one is taking propiltiouracil. Pathological studies revealed that 66% of cases were multinodular goiter and neoplasia was present in 13% of the patients. Other results were thymic pathology in 13% of cases (n=4), parathyroid cyst in 3,3% (n=1), and ectopic thyroid in 3,3% (n=1(.

Absence of transforming growth factor-beta type II receptor is associated with poorer prognosis in HER2-negative breast tumours.

BACKGROUND: The clinical relevance of transforming growth factor-beta (TGF-beta)-signalling pathway in breast carcinomas (BCs) remained elusive. This study aimed to evaluate the prognostic value of TGF-beta1 and transforming growth factor-beta type II receptor (TGF-betaRII) expression levels in tumour cells and their association with the established biomarkers in BC. PATIENTS AND METHODS: In 324 BC from patients with long-term follow-up, the TGF-beta1 and TGF-betaRII transcript and protein expression levels were assessed. RESULTS: TGF-beta1 and TGF-betaRII down-expression was significantly associated with BC. Negative TGF-beta1 and TGF-betaRII protein status was associated with the development of distant metastasis (P = 0.003 and P = 0.029, respectively). In multivariate analysis, TGF-beta1-positive tumours were associated with increased disease-free survival (DFS) [hazard ratio (HR) = 0.489, P = 0.003]. TGF-betaRII positivity was an independent prognostic factor for DFS (HR = 0.439, P = 0.001) and overall survival (OS) (HR = 0.409, P = 0.003) in human epidermal growth factor receptor-2 (HER2)-negative patients. Absence of TGF-beta1 and TGF-betaRII proteins in breast tumour cells was significantly associated with metastasis development. CONCLUSIONS: To the best of our knowledge, this is the first report indicating the relevance of HER2 status in discriminating TGF-betaRII as a prognostic marker for DFS and OS in human BC. These data indicate that TGF-betaRII protein analysis in tumour cells could be introduced in clinical practice as additional prognostic biomarker in HER2-negative BC.

[Pulmonary arteriovenous malformations in Rendu-Osler-Weber disease. Case report]. / Malformações arteriovenosas pulmonares: caso clínico no context de doenca de Rendu-Osler-Weber.

The authors report the clinical case of a 59 year-old woman who was incidentally diagnosed as having arteriovenous pulmonary malformation (AVPM). Through clinical history a diagnosis of Rendu-Osler-Weber disease (ROWD) was made, a disease manifesting itself through the existence of vascular malformations in the skin, mucosae and viscera (like the lungs, liver and brain). The surgical therapy offered to the patient had no complications. The essential aspects of pathofisiology, clinical manifestations and treatment of AVPM and ROWD are discussed.

[Surgery of pulmonary metastasis]. / Cirurgia das metáses pulmonares.

UNLABELLED: Between April 1993 and December 2007, 92 patients underwent the complete resection of pulmonary metastases, 47 mens (51.1%) and 45 womens (49.9%). 26 patients (27.2%) had solitary pulmonary metastasis, 25 (26.1%) multiple pulmonary metastasis and 41 (44.5%) patients presented bilateral pulmonary metastasis. 42 patients (45.6%) presented pulmonary metastasis recurrence, 14 patients (15.2%) a second recurrence and 6 patients (6.5%) a third pulmonary metastasis recurrence. 49 patients (53.2%) were reoperated. The mean cumulative survival after complete resection was 78.4 months (+/- 52.5 months). There were no operative mortality. CONCLUSION: Thys work demonstrated that every attempt should be made to completely ressect all clinically detected metastases. Complete resection of pulmonary metastasis, even in recurrent disease is compatible with long-term survival.

Survival patterns of Dekkera bruxellensis in wines and inhibitory effect of sulphur dioxide.

The wine spoilage yeast species Dekkera bruxellensis, after inoculation in red wines, displayed three survival patterns characterized by: i) initial lag phase followed by growth and sequential death; ii) initial death phase leading to reduced viable counts followed by growth and sequential death; and iii) death phase leading to complete loss of viability. These survival patterns were observed for the same strain in different dry red wine blends with 12% (v/v) ethanol and pH 3.50, in the absence of free sulphur dioxide. For the same wine blend, these patterns also varied with the tested strain. Under laboratory conditions the addition of 150 mg/l of potassium metabisulphite (PMB) to dry red wine with 12% (v/v) ethanol and pH 3.50 reduced initial cell counts by more than 6 logarithmic cycles, inducing full death within less than 24 h. Winery trials showed that D. bruxellensis blooms were only prevented in the presence of about 40 mg/l of free sulphur dioxide in dry red wine, with 13.8% (v/v) ethanol and pH 3.42, matured in oak barrels. These different amounts of PMB and sulphur dioxide corresponded to about 1 mg/l of molecular sulphur dioxide. Our results therefore demonstrate that the control of populations of D. bruxellensis growing in red wine can only be achieved under the presence of relatively high doses of molecular sulphur dioxide.

Gene expression profiling of clinical stages II and III breast cancer.

Clinical stage (CS) is an established indicator of breast cancer outcome. In the present study, a cDNA microarray platform containing 692 genes was used to identify molecular differences between CSII and CSIII disease. Tumor samples were collected from patients with CSII or CSIII breast cancer, and normal breast tissue was collected from women without invasive cancer. Seventy-eight genes were deregulated in CSIII tumors and 22 in CSII tumors when compared to normal tissue, and 20 of them were differentially expressed in both CSII and CSIII tumors. In addition, 58 genes were specifically altered in CSIII and expression of 6 of them was tested by real time RT-PCR in another cohort of patients with CSII or CSIII breast cancer and in women without cancer. Among these genes, MAX, KRT15 and S100A14, but not APOBEC3G or KRT19, were differentially expressed on both CSIII and CSII tumors as compared to normal tissue. Increased HMOX1 levels were detected only in CSIII tumors and may represent a molecular marker of this stage. A clear difference in gene expression pattern occurs at the normal-to-cancer transition; however, most of the differentially expressed genes are deregulated in tumors of both CS (II and III) compared to normal breast tissue.

Gene expression profiling of clinical stages II and III breast cancer

Clinical stage (CS) is an established indicator of breast cancer outcome. In the present study, a cDNA microarray platform containing 692 genes was used to identify molecular differences between CSII and CSIII disease. Tumor samples were collected from patients with CSII or CSIII breast cancer, and normal breast tissue was collected from women without invasive cancer. Seventy-eight genes were deregulated in CSIII tumors and 22 in CSII tumors when compared to normal tissue, and 20 of them were differentially expressed in both CSII and CSIII tumors. In addition, 58 genes were specifically altered in CSIII and expression of 6 of them was tested by real time RT-PCR in another cohort of patients with CSII or CSIII breast cancer and in women without cancer. Among these genes, MAX, KRT15 and S100A14, but not APOBEC3G or KRT19, were differentially expressed on both CSIII and CSII tumors as compared to normal tissue. Increased HMOX1 levels were detected only in CSIII tumors and may represent a molecular marker of this stage. A clear difference in gene expression pattern occurs at the normal-to-cancer transition; however, most of the differentially expressed genes are deregulated in tumors of both CS (II and III) compared to normal breast tissue.

[Cervical lymphadenopathy as first manifestation of a bronchogenic cyst]. / Linfadenopatias cervicais como primeira manifestação de um quisto broncogênico. Caso clinico.

A 26-year old woman was admitted with clinical and laboratory signs and symptoms of chronic infection and multiple cervical lymphadenopathy, whose pathological studies disclosed a non-specific inflammatory reaction. Anti-tuberculosis treatment was then started, with no significant improvement. Chest CT scans was compatible with a mediastinial tumor and RMN studies revealed the presence of an infected bronchogenic cyst, which was surgically and successfully removed with complete disappearance of the infection. The authors conclude by enhancing the fact that bronchogenic cysts may course with diverse clinical manifestations and should be included in differential diagnosis of mediastinal tumors.

[Surgery of mediastinal tumors: 11-years experience]. / Cirurgia dos tumores do mediastino: experiência de 11 anos.

Between 1993 and 2004, 70 operations on patients with the diagnosis of mediastinal tumors were performed in the Cardiothoracic Surgery Department of Santa Maria Hospital. In this study we did not include diagnostic surgery of mediastinal tumors. In 70 patients treated, 63 p (90%) had the tumor located in the anterior mediastinum, 21 p with thymoma, 6 p with lymphoma, 4 p with carcinoid tumors, 1 p with sarcoma and 1 p with adenocarcinoma. 3 patients had tumors located in the medium and 4 patients in the posterior mediastinum. We analysed the results of the mediastinal tumors surgery during the 11-years period. Also, we evaluated the clinical effects of the thymectomy in myasthenia gravis.