Epithelial-to-mesenchymal transition mediates docetaxel resistance and high risk of relapse in prostate cancer

Molecular characterization of radical prostatectomy specimens after systemic therapy may identify a gene expression profile for resistance to therapy. This study assessed tumor cells from patients with prostate cancer participating in a phase II neoadjuvant docetaxel and androgen deprivation trial to identify mediators of resistance. Transcriptional level of 93 genes from a docetaxel-resistant prostate cancer cell lines microarray study was analyzed by TaqMan low-density arrays in tumors from patients with high-risk localized prostate cancer (36 surgically treated, 28 with neoadjuvant docetaxel þ androgen deprivation). Gene expression was compared between groups and correlated with clinical outcome. VIM, AR and RELA were validated by immunohistochemistry. CD44 and ZEB1 expression was tested by immunofluorescence in cells and tumor samples. Parental and docetaxel-resistant castration-resistant prostate cancer cell lines were tested for epithelial-to-mesenchymal transition (EMT) markers before and after docetaxel exposure. Reversion of EMT phenotype was investigated as a docetaxel resistance reversion strategy. Expression of 63 (67.7%) genes differed between groups (P < 0.05), including genes related to androgen receptor, NF-k B transcription factor, and EMT. Increased expression of EMT markers correlated with radiologic relapse. Docetaxel-resistant cells had increased EMT and stem-like cell markers expression. ZEB1 siRNA transfection reverted docetaxel resistance and reduced CD44 expression in DU-145R and PC-3R. Before docetaxel exposure, a selected CD44 þ subpopulation of PC-3 cells exhibited EMT phenotype and intrinsic docetaxel resistance; ZEB1/CD44 þ subpopulations were found in tumor cell lines and primary tumors; this correlated with aggressive clinical behavior. This study identifies genes potentially related to chemotherapy resistance and supports evi-dence of the EMT role in docetaxel resistance and adverse clinical behavior in early prostate cancer.

Modeling dynamic behavior in tilting pad gas journal bearings

Thisresearch deals with the dynamic modeling of gas lubricated tilting pad journal bearings provided with spring supported pads, including experimental verification of the computation. On the basis of a mathematical model of a film bearing, a computer program has been developed, which can be used for the simulation of a special type of tilting pad gas journal bearing supported by a rotary spring under different loading conditions time dependently (transient running conditions due to geometry variations in time externally imposed). On the basis of literature, different transformations have been used in the model to achieve simpler calculation. The numerical simulation is used to solve a non-stationary case of a gasfilm. The simulation results were compared with literature results in a stationary case (steady running conditions) and they were found to be equal. In addition to this, comparisons were made with a number of stationary and non-stationary bearing tests, which were performed at Lappeenranta University of Technology using bearings designed with the simulation program. A study was also made using numerical simulation and literature to establish the influence of the different bearing parameters on the stability of the bearing. Comparison work was done with literature on tilting pad gas bearings. This bearing type is rarely used. One literature reference has studied the same bearing type as that used in LUT. A new design of tilting pad gas bearing is introduced. It is based on a stainless steel body and electron beam welding of the bearing parts. It has good operation characteristics and is easier to tune and faster to manufacture than traditional constructions. It is also suitable for large serial production.

Obstructive apneas induce early activation of mesenchymal stem cells and enhancement of endothelial wound healing

Background: The aim was to test the hypothesis that the blood serum of rats subjected to recurrent airway obstructions mimicking obstructive sleep apnea (OSA) induces early activation of bone marrow-derived mesenchymal stem cells (MSC) and enhancement of endothelial wound healing. Methods: We studied 30 control rats and 30 rats subjected to recurrent obstructive apneas (60 per hour, lasting 15 s each, for 5 h). The migration induced in MSC by apneic serum was measured by transwell assays. MSC-endothelial adhesion induced by apneic serum was assessed by incubating fluorescent-labelled MSC on monolayers of cultured endothelial cells from rat aorta. A wound healing assay was used to investigate the effect of apneic serum on endothelial repair. Results: Apneic serum showed significant increase in chemotaxis in MSC when compared with control serum: the normalized chemotaxis indices were 2.20 +- 0.58 (m +- SE) and 1.00 +- 0.26, respectively (p < 0.05). MSC adhesion to endothelial cells was greater (1.75 +- 0.14 -fold; p < 0.01) in apneic serum than in control serum. When compared with control serum, apneic serum significantly increased endothelial wound healing (2.01 +- 0.24 -fold; p < 0.05). Conclusions: The early increases induced by recurrent obstructive apneas in MSC migration, adhesion and endothelial repair suggest that these mechanisms play a role in the physiological response to the challenges associated to OSA.

Immunomodulatory effect of mesenchymal stem cells on B cells

The research on T cell immunosuppression therapies has attracted most of the attention in clinical transplantation. However, B cells and humoral immune responses are increasingly acknowledged as crucial mediators of chronic allograft rejection. Indeed, humoral immune responses can lead to renal allograft rejection even in patients whose cell-mediated immune responses are well controlled. On the other hand, newly studied B cell subsets with regulatory effects have been linked to tolerance achievement in transplantation. Better understanding of the regulatory and effector B cell responses may therefore lead to new therapeutic approaches. Mesenchymal stem cells (MSC) are arising as a potent therapeutic tool in transplantation due to their regenerative and immunomodulatory properties.The research on MSCs has mainly focused on their effects onT cells and although data regarding the modulatory effects of MSCs on alloantigen-specific humoral response in humans is scarce, it has been demonstrated that MSCs significantly affect B cell functioning. In the present review we will analyze and discuss the results in this field.

The Human Mesenchymal Stromal Cell-Derived Osteocyte Capacity to Modulate Dendritic Cell Functions Is Strictly Dependent on the Culture System.

In vitro differentiation of mesenchymal stromal cells (MSC) into osteocytes (human differentiated osteogenic cells, hDOC) before implantation has been proposed to optimize bone regeneration. However, a deep characterization of the immunological properties of DOC, including their effect on dendritic cell (DC) function, is not available. DOC can be used either as cellular suspension (detached, Det-DOC) or as adherent cells implanted on scaffolds (adherent, Adh-DOC). By mimicking in vitro these two different routes of administration, we show that both Det-DOC and Adh-DOC can modulate DC functions. Specifically, the weak downregulation of CD80 and CD86 caused by Det-DOC on DC surface results in a weak modulation of DC functions, which indeed retain a high capacity to induce T-cell proliferation and to generate CD4(+)CD25(+)Foxp3(+) T cells. Moreover, Det-DOC enhance the DC capacity to differentiate CD4(+)CD161(+)CD196(+) Th17-cells by upregulating IL-6 secretion. Conversely, Adh-DOC strongly suppress DC functions by a profound downregulation of CD80 and CD86 on DC as well as by the inhibition of TGF-β production. In conclusion, we demonstrate that different types of DOC cell preparation may have a different impact on the modulation of the host immune system. This finding may have relevant implications for the design of cell-based tissue-engineering strategies.

Impaired aldehyde dehydrogenase 1 subfamily member 2A-dependent retinoic acid signaling is related with a mesenchymal-like phenotype and an unfavorable prognosis of head and neck squamous cell carcinoma.

BACKGROUND: An inverse correlation between expression of the aldehyde dehydrogenase 1 subfamily A2 (ALDH1A2) and gene promoter methylation has been identified as a common feature of oropharyngeal squamous cell carcinoma (OPSCC). Moreover, low ALDH1A2 expression was associated with an unfavorable prognosis of OPSCC patients, however the causal link between reduced ALDH1A2 function and treatment failure has not been addressed so far. METHODS: Serial sections from tissue microarrays of patients with primary OPSCC (n = 101) were stained by immunohistochemistry for key regulators of retinoic acid (RA) signaling, including ALDH1A2. Survival with respect to these regulators was investigated by univariate Kaplan-Meier analysis and multivariate Cox regression proportional hazard models. The impact of ALDH1A2-RAR signaling on tumor-relevant processes was addressed in established tumor cell lines and in an orthotopic mouse xenograft model. RESULTS: Immunohistochemical analysis showed an improved prognosis of ALDH1A2(high) OPSCC only in the presence of CRABP2, an intracellular RA transporter. Moreover, an ALDH1A2(high)CRABP2(high) staining pattern served as an independent predictor for progression-free (HR: 0.395, p = 0.007) and overall survival (HR: 0.303, p = 0.002), suggesting a critical impact of RA metabolism and signaling on clinical outcome. Functionally, ALDH1A2 expression and activity in tumor cell lines were related to RA levels. While administration of retinoids inhibited clonogenic growth and proliferation, the pharmacological inhibition of ALDH1A2-RAR signaling resulted in loss of cell-cell adhesion and a mesenchymal-like phenotype. Xenograft tumors derived from FaDu cells with stable silencing of ALDH1A2 and primary tumors from OPSCC patients with low ALDH1A2 expression exhibited a mesenchymal-like phenotype characterized by vimentin expression. CONCLUSIONS: This study has unraveled a critical role of ALDH1A2-RAR signaling in the pathogenesis of head and neck cancer and our data implicate that patients with ALDH1A2(low) tumors might benefit from adjuvant treatment with retinoids.

Étude du rapport entre les manifestations d'agitation et l'inconfort de la personne âgée atteinte de démence (pad) au moment de la toilette en milieu hospitalier spécialise

Les personnes âgées atteintes de démence (PAD) souffrent de troubles importants de la communication associés à des manifestations psychologiques et comportementales comme l'agitation. La plupart de ces personnes hospitalisées sont en outre atteintes de comorbidités physiques ou psychologiques pouvant provoquer de l'inconfort. Ce travail décrit la fréquence des comportements d'agitation et de l'inconfort des PAD durant les soins d'hygiène et vérifie si ces deux phénomènes sont associés durant la toilette. La mise en perspective infirmière des résultats de l'étude est effectuée à partir des écrits scientifiques et du cadre théorique du confort de Kolcaba.

Use of Bichat"s buccal fat pad for the sealing of orosinusal communications. A presentation of 8 cases

Objectives: To determine the efficacy of the pediculate flap with the buccal fat pad in the sealing of orosinusal communications, describe the surgical technique used, and report the main complications. Patients and method: A retrospective study was made of 8 patients seen in the Service of Oral Surgery of the University of Barcelona Dental Clinic (Spain) for the treatment of orosinusal communications between the years 2007 and 2009. In all cases a pediculate flap with the buccal fat pad was used to solve the problem. Results: All of the orosinusal communications were successfully resolved with this technique. The immediate postoperative complications were pain (37.5%), inflammation (37.5%), edema (32.5%), trismus (37.5%), halitosis (14.3%), suppuration (12.5%) and rhinorrhea (12.5%). Conclusions: The use of Bichat"s buccal fat pad is not regarded as the technique of choice for sealing small to medium sized orosinusal communications. However, in the case of large communications, it is a good option, and the results obtained are optimum.

Epithelial-to-mesenchymal transition mediates docetaxel resistance and high risk of relapse in prostate cancer

Molecular characterization of radical prostatectomy specimens after systemic therapy may identify a gene expression profile for resistance to therapy. This study assessed tumor cells from patients with prostate cancer participating in a phase II neoadjuvant docetaxel and androgen deprivation trial to identify mediators of resistance. Transcriptional level of 93 genes from a docetaxel-resistant prostate cancer cell lines microarray study was analyzed by TaqMan low-density arrays in tumors from patients with high-risk localized prostate cancer (36 surgically treated, 28 with neoadjuvant docetaxel þ androgen deprivation). Gene expression was compared between groups and correlated with clinical outcome. VIM, AR and RELA were validated by immunohistochemistry. CD44 and ZEB1 expression was tested by immunofluorescence in cells and tumor samples. Parental and docetaxel-resistant castration-resistant prostate cancer cell lines were tested for epithelial-to-mesenchymal transition (EMT) markers before and after docetaxel exposure. Reversion of EMT phenotype was investigated as a docetaxel resistance reversion strategy. Expression of 63 (67.7%) genes differed between groups (P < 0.05), including genes related to androgen receptor, NF-k B transcription factor, and EMT. Increased expression of EMT markers correlated with radiologic relapse. Docetaxel-resistant cells had increased EMT and stem-like cell markers expression. ZEB1 siRNA transfection reverted docetaxel resistance and reduced CD44 expression in DU-145R and PC-3R. Before docetaxel exposure, a selected CD44 þ subpopulation of PC-3 cells exhibited EMT phenotype and intrinsic docetaxel resistance; ZEB1/CD44 þ subpopulations were found in tumor cell lines and primary tumors; this correlated with aggressive clinical behavior. This study identifies genes potentially related to chemotherapy resistance and supports evi-dence of the EMT role in docetaxel resistance and adverse clinical behavior in early prostate cancer.

The maximum voltage drop in an on-chip power distribution network: analysis of square, triangular and hexagonal power pad arrangements

A mathematical model of the voltage drop which arises in on-chip power distribution networks is used to compare the maximum voltage drop in the case of different geometric arrangements of the pads supplying power to the chip. These include the square or Manhattan power pad arrangement, which currently predominates, as well as equilateral triangular and hexagonal arrangements. In agreement with the findings in the literature and with physical and SPICE models, the equilateral triangular power pad arrangement is found to minimize the maximum voltage drop. This headline finding is a consequence of relatively simple formulas for the voltage drop, with explicit error bounds, which are established using complex analysis techniques, and elliptic functions in particular.

Therapeutic Properties of Superoxide Dismutase 3 and Mesenchymal Stromal Cells in Peripheral Ischemia

The aim of this study was to characterize the cellular mechanisms leading to the beneficial effect of anti-oxidative gene therapy and pro-angiogenic stem cell therapy in acute peripheral ischemia. Post-ischemic events aim to re-establish tissue blood perfusion, to clear cellular debris, and to regenerate lost tissue by differentiation of satellite cells into myoblasts. Although leukocytes have an essential role in clearing cellular debris and promoting angiogenesis, they also contribute to tissue injury through excessive ROS production. First, we investigated the therapeutic properties of extracellular superoxide dismutase (SOD3) gene transfer. SOD3 was shown to reduce oxidative stress, to normalize glucose metabolism, and to enhance cell proliferation in the ischemic muscle. Analysis of the mitogenic Ras-Erk1/2 pathway showed SOD3 mediated induction offering a plausible explanation for enhanced cell proliferation. In addition, SOD3 reduced NF-κB activity by enhancing IκBα expression thus leading to reduced expression of inflammatory cytokines and adhesion molecules with consequent reduction in macrophage infiltration. Secondly, we sought to determine the fate and the effect of locally transplanted mesenchymal stem/stromal cells (MSCs) in acute ischemia. We showed that a vast majority of the transplanted cells are cleared from the injury site within 24 hours after local transplantation. Despite rapid clearance, transplantation was able to temporarily promote angiogenesis and cell proliferation in the muscle. Lack of graft-derived growth factor expression suggests other than secretory function to mediate this observed effect. In conclusion, both SOD3 and MSCs could be utilized to alleviate peripheral ischemia induced tissue injury. We have described a previously unidentified growth regulatory role for SOD3, and suggest a novel mechanism whereby transplanted MSCs enhance the reparative potential of the recipient tissue through physical contacts.

Peritônio bovino conservado na correção de hérnia ventral em ratos: uma alternativa para tela cirúrgica biológica

OBJETIVO: Verificar a possibilidade de implantação e a capacidade de resistência tênsil do peritônio parietal bovino como tela cirúrgica na correção de hérnia ventral em um modelo animal de experimentação. MÉTODO: Utilizando 57 ratos machos Wistar, comparou-se o implante do peritônio bovino com a tela de polipropileno na correção de um defeito provocado na parede abdominal do animal. Após sete (sub-grupo A) e 28 (sub-grupo B) dias de observação, as peças foram retiradas e procedeu-se a avaliação da resistência à tração em Máquina Universal de Ensaios. Um grupo sem implante de material protético foi utilizado como controle nos testes de força tênsil. Os testes de Mann-Whitney e de Kruskal-Wallis foram utilizados e estabeleceu-se em 0,05 o nível para rejeição da hipótese de nulidade. RESULTADOS: Os testes de resistência à tração, com valores expressos em Newton, não mostraram diferenças estatísticas entre os grupos estudados, tanto no 7º quanto no 28º dia de pós-operatório, e ambos foram menos resistentes que a parede abdominal normal (p = 0,003). CONCLUSÃO: O peritônio parietal bovino apresentou resistência tênsil semelhante a da tela de polipropileno em um modelo de correção de hérnia ventral em ratos.

Reparo intraperitoneal de defeitos da parede ventral do abdome com telas de poliéster com colágeno e polipropileno com ácido poliglicólico

OBJETIVO: Avaliar a incorporação de telas de poliéster revestido em uma de suas faces por colágeno (Parietex, Covidien) e polipropileno recoberto por ácido poliglicólico (Optilene Mesh Elastic e Safil, BBD Aesculap) no reparo de defeitos da parede ventral de coelhos avaliando a cicatrização no aspecto macroscópico, o depósito de colágeno e a imunomarcação tecidual pelos anticorpos MMP-1, MMP-8 e MMP-13. MÉTODOS: Utilizaram-se 16 coelhos, divididos em dois grupos de oito animais, avaliados após eutanásia após 30 e 60 dias de pós-operatório. Os animais foram submetidos à realização de dois defeitos simétricos na parede ventral do abdome, à direita e esquerda da linha alba, que compreendendo todos os folhetos musculares e o peritônio. O reparo dos defeitos foi realizado mediante implante intraperitoneal de dois modelos diferentes de telas. Utilizou-se a tela de poliéster revestido com camada protetora de colágeno (grupo controle) e a tela de polipropilene revestido com malha de ácido poliglicólico (manufaturacao própria, grupo de experimentacao). A avaliacao constou de aspectos clínicos, achados macroscópicos, análise dos colágenos tipos I/III e avaliação imunoistoquímica de metaloproteinases. RESULTADOS: Os resultados da avaliacao clínica e os parâmetros macroscópicos foram semelhantes entre os grupos. 50% dos animais do grupo Parietex tiveram ausência de aderencias intraperitoneais a no 30° dia de pós-operatrório. Em ambos os grupos observou-se reducao das aderências entre o 30° e o 60° dias de pós-operatório, contudo sem diferenca estatística. As aderências observadas foram classificadas principalmente de frouxas. Nao se observou a ocorrencia de complicacoes envolvendo vísceras intraabdominais. No Grupo Parietex houve a ocorrência de formacao de ulceracao da pele que recobria a tela em quatro animais, em comparacao com um no grupo de experimentacao. No Grupo Parietex foi observada uma insuficiencia de reparo após 60 dias. Quanto ao depósito do colágeno tipos I e III, nao houve diferenca significativa entre os grupos. Os resultados da imunoistoquímica referentes aos anticorpos MMP-1 e MMP-8 também não demonstraram diferença significativa entre as telas. CONCLUSÃO: As duas telas pesquisadas obtiveram resultados semelhantes tanto nos aspectos macro como nos microscópicos, podendo ser consideradas semelhantes quanto ao reparo de defeitos cirúrgicos da parede ventral do abdome em coelhos.

Correlação entre as queixas de incontinência urinária de esforço e o pad test de uma hora em mulheres na pós-menopausa

OBJETIVO: correlacionar as queixas de incontinência urinária de esforço e os resultados da aplicação do pad test de uma hora em mulheres na pré e pós-menopausa. MÉTODOS: estudo transversal, composto por 60 voluntárias na pós-menopausa, divididas em dois grupos: um com 34 mulheres com queixa de perda involuntária de urina aos esforços, outro com 26 mulheres sem queixas de perda de urina aos esforços. Há também a presença de um Grupo Controle composto por 15 mulheres na pré-menopausa, com ciclo menstrual normal e sem queixas urinárias. Todas as mulheres foram avaliadas quanto à clínica e laboratorialmente, e submetidas ao pad test por uma hora. A paciente foi considerada incontinente quando o peso do absorvente após o teste foi maior do que 1 g. Os resultados obtidos foram submetidos à estatística descritiva, ao teste paramétrico ANOVA, ao pós-teste de Turkey e à correlação de Pearson. RESULTADOS: todas as mulheres na pós-menopausa apresentaram incontinência urinária de esforço durante o pad test, tanto as que referiram perda urinária (4 g), como as sem perda urinária prévia (3,5 g). Nessas mulheres, observou-se uma forte correlação das perdas de urina com o tempo de menopausa (r=0,8; p<0,01) e com o índice de massa corpórea (IMC) (r=0,7; p= 0,01). As mulheres na pré-menopausa mantiveram-se continentes durante o pad test (0,4 g). CONCLUSÕES: os resultados obtidos com a aplicação do pad test de uma hora mostraram que todas as mulheres na pós-menopausa apresentavam incontinência urinária de esforço, inclusive aquelas que não apresentavam queixa de perda de urina aos esforços. Essa perda de urina correlaciona-se com o tempo de menopausa e com o IMC.