bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic role of p‐mTOR expression in cancer tissues and metastatic lymph nodes in pT2b gastric cancer
Authors: An Ji Yeong , Kim Kyoung Mee , Choi Min Gew , Noh Jae Hyung , Sohn Tae Sung , Bae Jae Moon , Kim Sung
Published: September 2009

Despite the great interest in mammalian target of rapamycin (mTOR) as a potential anticancer therapy target, the prognostic role of mTOR in gastric cancer has not been elucidated. In this study, we investigated mTOR expression in gastric cancer tissues and in metastatic lymph nodes and examined its association with clinical outcome. A total of 290 patients with pT2b gastric cancer were enrolled in this study. Patients were divided into 3 groups according to metastatic lymph node status: Group 1 contained 96 patients without lymph node metastasis, Group 2 contained 102 patients with a few (1–2) metastatic lymph nodes and Group 3 contained 92 patients with extensive (>16) lymph node metastasis. Phosphorylated mTOR expression was determined immunohistochemically using tissue microarrays. p‐mTOR expression was observed in 36.5% of the gastric cancer tissues in Group 1, 39.2% in Group 2 and 60.9% in Group 3. A significant correlation was found between p‐mTOR expression in gastric cancer tissues and in metastatic lymph nodes. The Borrmann type in Group 1, perineural invasion and p‐mTOR expression in metastatic lymph nodes in Group 2 and p‐mTOR expression in metastatic lymph nodes in Group 3 were found to be independent prognostic factors of disease‐free survival. The 5‐year disease free survival rate of Group 2 patients was 84.4% in negative p‐mTOR and 66.1% in positive p‐mTOR expression in metastatic lymph nodes (p = 0.015). The 5‐year disease free survival rate of Group 3 patients was 37.3% in negative p‐mTOR and 14.9% in positive p‐mTOR expression in metastatic lymph nodes (p = 0.037). There was a linear correlation between the rate of tumor recurrence and mTOR expression scores in metastatic lymph nodes. In pT2b gastric cancer, p‐mTOR expression in gastric cancer is associated with the extent of lymph node metastasis, and p‐mTOR expression in metastatic lymph nodes is correlated with poor disease‐free survival. mTOR may harbor significant potential for a prognostic biomarker and therapeutic target for gastric cancer treatment.
The influence of a period of caloric restriction due to the Dutch famine on breast density
Authors: Haars Gerco , van Gils Carla H. , Elias Sjoerd G. , Lokate Mariëtte , van Noord Paul A.H. , Peeters Petra HM
Published: September 2009

Recently, we showed that short, intense caloric restriction due to the Dutch famine increased breast cancer risk in women and influenced the hormonal‐ and IGF‐system. These systems may also affect breast density, which is one of the strongest risk factors for breast cancer. We examined the influence of the Dutch famine on breast density, using mammograms of 1,035 women. Breast size, dense and non‐dense tissue and the relative density were measured on a continuous scale. Mean density was compared between three groups of ascending levels of famine‐exposure. Results were adjusted for known determinants of breast density and stratified by age at exposure. There were no overall, significant differences by exposure. In unexposed compared to severely exposed women, means varied from 124 cm2 to 121 cm2 (ptrend = 0.50) for breast size, from 23.4 to 21.8 cm2 (ptrend = 0.48) for amount of dense tissue, from 87.7 to 85.4 cm2 (ptrend = 0.55) for non‐dense tissue and from 22.8 to 22.3% (ptrend = 0.78) for relative density. Only among women who were younger than 10 years during the famine was the amount of non‐dense tissue significantly lower with higher exposure, with 53.1 cm2 for severely exposed compared to 77.8 cm2 (ptrend = 0.03) for unexposed. This group also appeared to have smaller breasts with more absolute and relative density, but not statistically significant. We observed no overall effect on breast density in women severely exposed to a short, intense caloric restriction. However, in women exposed before puberty, the non‐dense area was smaller and density tended to be higher.
Cutaneous type adult T‐cell leukemia/lymphoma is a characteristic subtype and includes erythema/papule and nodule/tumor subgroups
Authors: Miyata Tomoko , Yonekura Kentaro , Utsunomiya Atae , Kanekura Takuro , Nakamura Shigeo , Seto Masao
Published: September 2009

We first analyzed the genomic profile of cutaneous type adult T‐cell leukemia/lymphoma (ATLL) in an attempt to clarify its clinical and biological characteristics. Genomic gains of 1p, 7q and 18q and loss of 13q were frequently detected. Gain of 1p36.33‐32 or loss of 13q33.1‐3 indicated poor prognosis. Among cases with generalized lesions, erythema/papule or nodule/tumor cases showed a distinct genomic profile, indicating that these 2 groups were biologically different and developed via different genetic pathways. Furthermore, cases with generalized nodule/tumor lesions tended to progress to aggressive ATLL.
Microsomal prostaglandin E synthase‐1 inhibition blocks proliferation and enhances apoptosis in oesophageal adenocarcinoma cells without affecting endothelial prostacyclin production
Authors: Beales Ian L.P. , Ogunwobi Olorunseun O.
Published: September 2009

Prostaglandins are important in the progression of various gastrointestinal cancers including oesophageal adenocarcinoma (OAC). Cyclo‐oxygenase (COX)‐2 inhibitors reduce OAC prostaglandin production but also have potentially detrimental effects on vascular endothelial function by reducing prostacyclin production and increasing the risk of cardiovascular events. We have examined the effects of inhibiting microsomal prostaglandin synthase‐1 (mPGES‐1), the enzyme downstream of COX‐2 in the prostaglandin synthetic cascade. In OAC cells, reduction of mPGES‐1 with RNA interference blocked PGE2 production, inhibited serum‐induced proliferation and enhanced apoptosis in the COX‐2 expressing cell lines (OE33 and FLO) but had no effect in COX‐2 deficient BIC‐1 cells. Three different methods of inhibiting mPGES‐1 (RNA interference, a novel small molecule inhibitor and the endogenous inhibitor 15‐deoxy‐Δ12,14‐PGJ2) also blocked leptin induced mPGES‐1 expression and PGE2 production and abolished the leptin‐induced proliferative and anti‐apoptotic effects in OE33 cells, without affecting COX‐2 expression. The anti‐proliferative effects were equivalent to those produced by COX‐2 inhibitory concentrations of celecoxib and NS‐398. However, unlike the two COX‐2 inhibitors, mPGES‐1 inhibition did not reduce endothelial prostacyclin production. In contrast to the effects of the COX‐2 inhibitor celecoxib, mPGES‐1 inhibition had no effects on Akt kinase activity in OAC cells. We conclude that inhibition of mPGES‐1 has potentially beneficial effects in OAC without the potentially detrimental effects on vascular endothelial prostacyclin synthesis. We have also confirmed that celecoxib has anticancer actions separate from the inhibition of COX‐2. Inhibition of mPGES‐1 may be therapeutically useful in the treatment and prevention of OAC.
Detection of Merkel cell polyomavirus (MCPyV) in Merkel cell carcinoma cell lines: Cell morphology and growth phenotype do not reflect presence of the virus
Authors: Fischer Nicole , Brandner Johanna , Fuchs Franca , Moll Ingrid , Grundhoff Adam
Published: September 2009

The recently discovered human polyomavirus (MCPyV) is frequently found in Merkel cell carcinoma (MCC) tissue and is believed to be causally linked to MCC pathogenesis. While cell lines established from MCC represent a valuable tool to study the contribution of MCPyV to MCC pathogenesis, hitherto only 1 MCPyV‐positive line has been described. We have analyzed 7 MCC cell lines for the presence, integration pattern and copy number of MCPyV. In 5 cell lines, MCPyV specific sequences were detected. In 3 of these lines, multiple copies of viral genomes per cell were detected, and sequencing of PCR amplificates identified distinct mutations predicted to lead to the expression of a truncated large T‐Antigen (LT‐Ag). In 1 cell line, clonal integration of concatamerized viral genomes was confirmed by Southern Blotting. MCC cell lines are conventionally categorized as “classic” or “variant” and further divided into 4 subtypes, based on expression of neuroendocrine markers and morphology. While it has been suggested that the presence of MCPyV might promote a classic phenotype, such a notion is not supported by our data. Instead, we find MCPyV‐positive as well as ‐negative lines of the classic variety, indicating that the distinguishing features are either inherently independent of viral infection or have become so in the course of tumorigenesis and/or cell line establishment. We therefore suggest a novel classification scheme based on MCPyV presence, integration patterns and T‐Ag mutations. The cell lines described here extend the repertoire of available MCPyV‐positive MCC‐lines and should aid in the elucidation of the role of MCPyV in the pathogenesis of MCC.
Molecular characterization of human homologs of yeast MOB1
Authors: Chow Annabelle , Hao Yawei , Yang Xiaolong
Published: September 2009

MOB (Mps one binder) was originally identified in yeast as a regulator of mitotic exit and cytokinesis, and was later identified as a tumor suppressor and a component of an emerging Hippo‐LATS tumor suppressor pathway in Drosophila (D). So far, 7 human homologs of yeast MOB (hMOB1A, 1B, 2A, 2B, 2C, 3, 4) have been identified. Although hMOB1A/B has been extensively studied, the biological features of other hMOBs are largely unknown. In addition, while hMOB1 has been reported to interact with and activate LATS (Large tumor suppressor)/Warts tumor suppressor, the functional significance of this is unknown. In this study, we have characterized, for the first time, the cellular and biochemical function of all human MOBs. By examining hMOB mRNAs expression in various human tissues, we found that hMOBs demonstrated different expression patterns. Further biochemical characterization of hMOBs showed that only hMOB1A and hMOB1B interact with both LATS1 and LATS2 in vitro and in vivo. Significantly, we have discovered that overexpression of hMOB1 in human cancer cells activated LATS activity and inhibited cell proliferation or caused apoptosis while hMOB1, targeting the plasma membrane, led to a more significant phenotype. Reciprocally, short‐hairpin (sh) RNA‐mediated suppression of hMOB1 causes increased cell proliferation. Our findings provided evidence that hMOB1A and hMOB1B are 2 LATS‐binding proteins that may function as tumor suppressors in human cancer cells.
Loss of FBXW7, a cell cycle regulating gene, in colorectal cancer: Clinical significance
Authors: Iwatsuki Masaaki , Mimori Koshi , Ishii Hideshi , Yokobori Takehiko , Takatsuno Yasushi , Sato Tetsuya , Toh Hiroyuki , Onoyama Ichiro , Nakayama Keiichi I. , Baba Hideo , Mori Masaki
Published: September 2009

This study focused on a cell cycle regulatory gene, FBXW7, which ubiquitinates c‐Myc and cyclin E and promotes exit from the cell cycle. We determined the expression level of FBXW7 in colorectal cancer (CRC) cases, correlated those values with clinicopathologic features, and characterized the molecular mechanism of reduced expression of FBXW7 in CRC cells in vitro. FBXW7 mRNA and protein expression were evaluated in 93 CRC cases. Using CGH array, the copy number aberrations of the flanking region of FBXW7 were evaluated in another 130 CRC specimens. In vitro analysis of FBXW7 gene silencing in CRC cells was conducted. FBXW7 mRNA expression was significantly lower in tumor tissues than the corresponding normal tissues. The low FBXW7 expression group showed a significantly poorer prognosis than patients in the high expression group. A concordant relationship was observed between the incidence of FBXW7 repression and the genetic alteration. The incidence of genetic alteration was associated with the stage of disease progression. In vitro, FBXW7‐specific siRNA enhanced expression of c‐MYC and cyclin E proteins and up‐regulated cell proliferation. Genetic alterations in tumors led to the loss of FBXW7 expression and increased cell proliferation. FBXW7 expression provides a prognostic factor for patients with CRC.
Significance of Plk1 regulation by miR‐100 in human nasopharyngeal cancer
Authors: Shi Wei , Alajez Nehad M. , Bastianutto Carlo , Hui Angela B.Y. , Mocanu Joseph D. , Ito Emma , Busson Pierre , Lo Kwok‐Wai , Ng Raymond , Waldron John , O'Sullivan Brian , Liu Fei‐Fei
Published: September 2009

Polo‐like kinase 1 (Plk1) is a critical regulator of many stages of mitosis; increasing evidence indicates that Plk1 overexpression correlates with poor clinical outcome, yet its mechanism of regulation remains unknown. Hence, a detailed evaluation was undertaken of Plk1 expression in human nasopharyngeal cancer (NPC), the cellular effects of targeting Plk1 using siRNA in combination with ionizing radiation (RT) and potential upstream microRNAs (miRs) that might regulate Plk1 expression. Using immunohistochemistry, Plk1 was observed to be overexpressed in 28 of 40 (70%) primary NPC biopsies, which in turn was associated with a higher likelihood of recurrence (p = 0.018). SiPlk1 significantly inhibited Plk1 mRNA and protein expression, and decreased Cdc25c levels in NPC cell lines. This depletion resulted in cytotoxicity of C666‐1 cells, enhanced by the addition of RT, mediated by G2/M arrest, increased DNA double‐strand breaks, apoptosis, and caspase activation. Immunofluorescence demonstrated that the G2/M arrest was associated with aberrant spindle formation, leading to mitotic arrest. In vivo, transfection of C666‐1 cells and systemic delivery of siPlk1 decreased tumour growth. MicroRNA‐100 (miR‐100) was predicted to target Plk1 mRNA, which was indeed underexpressed in C666‐1 cells, inversely correlating with Plk1 expression. Using luciferase constructs containing the 3′‐UTR of Plk1 sequence, we document that miR‐100 can directly target Plk1. Hence, our data demonstrate for the first time that underexpressed miR‐100 leads to Plk1 overexpression, which in turn contributes to NPC progression. Targeting Plk1 will cause mitotic catastrophe, with significant cytotoxicity both in vitro and in vivo, underscoring the important therapeutic opportunity of Plk1 in NPC.
Enhancement of tumor‐specific T‐cell responses by transcatheter arterial embolization with dendritic cell infusion for hepatocellular carcinoma
Authors: Mizukoshi Eishiro , Nakamoto Yasunari , Arai Kuniaki , Yamashita Tatsuya , Mukaida Naofumi , Matsushima Kouji , Matsui Osamu , Kaneko Shuichi
Published: September 2009

Transcatheter arterial embolization (TAE) destroys a tumor by the induction of necrosis and/or apoptosis and causes inflammation with cytokine production, which may favor immune activation and presentation of tumor‐specific antigens. In the current study, we attempted to identify the effect of TAE on tumor‐specific T‐cell responses and the additional effect of dendritic cell (DC) infusion performed during TAE. The prevalence of tumor antigen‐specific T cells was determined by interferon‐γ enzyme‐linked immunospot analysis using alpha‐fetoprotein (AFP) and tumor antigen‐derived peptides in 20 and 13 patients with hepatocellular carcinoma (HCC) who received TAE and TAE with DC infusion, respectively. The increased frequency of AFP‐specific T cells was observed in 6 of 20 patients after TAE. It was observed more frequently in patients with DC infusion than in those with TAE alone. However, tumor recurrence was not completely prevented in patients albeit displayed enhanced immune responses. The evidence that the enhanced immune responses were transient and attenuated within 3 months was provided in time‐course analysis. In conclusion, TAE with DC infusion enhances the tumor‐specific immune responses more effectively than TAE alone. Although the effect is not sufficient to prevent HCC recurrence, these results may contribute to the development of novel immunotherapeutic approach for HCC.
Double impact on p‐glycoprotein by statins enhances doxorubicin cytotoxicity in human neuroblastoma cells
Authors: Sieczkowski Evelyn , Lehner Claudia , Ambros Peter F. , Hohenegger Martin
Published: September 2009

The development of multidrug resistance (MDR) is a major problem during cancer treatment. Drug efflux via ATP‐binding cassette (ABC) transporters is the main mechanism responsible for resistance to chemotherapeutics. We have recently observed that statins enhance susceptibility to doxorubicin‐induced apoptosis in human rhabdomyosarcoma cells, which is now also observed in human SH‐SY5Y neuroblastoma cells. We have therefore investigated the ABC transporter activity to confirm a possible inhibition by statins in SH‐SY5Y cells. Indeed, simvastatin directly inhibited dye transport at equimolar concentrations of the ABC transporter inhibitor, verapamil. Making use of the fluorescence behavior of doxorubicin the accumulation of anthracycline was monitored in real‐time confocal microscopy. The intracellular doxorubicin accumulation was immediately enhanced by statins in SH‐SY5Y cells and also in a MYCN‐amplified neuroblastoma cell line STA‐NB‐10. The heavily glycosylated P‐glycoprotein (ABCB1, P‐gp) transporter appeared as a 180‐and 140‐kDa species. Atorvastatin and simvastatin reduced the 180‐kDa form of P‐gp, but not verapamil. Thereby the fully glycosylated species is shifted to the core glycosylated species (140 kDa), which was only seen at statin exposure times longer than 24 hr. The functional importance of glycosylation of the transporter was highlighted by exogenous application of N‐glycosidase F, which was sufficient to enhance doxorubicin accumulation. Hence, these novel findings of statins' dual impact on P‐gp have clinical implications. The enhanced intracellular accumulation of chemotherapeutics or other ABC transporter substrates in the presence of statins may represent a novel concept to overcome MDR in cancer therapy and improve drug safety.
Mortalin inhibitors sensitize K562 leukemia cells to complement‐dependent cytotoxicity
Authors: Pilzer David , Saar Moran , Koya Keizo , Fishelson Zvi
Published: September 2009

Mortalin, the mitochondrial hsp70, is a vital constitutively expressed heat shock protein. Its elevated expression has been correlated with malignant transformation and poor cancer prognosis. Cancer cells exhibit increased resistance to complement‐dependent cytotoxicity, partly due to their capacity to eliminate the complement membrane attack complex (MAC) from their cell surface. As we have previously reported, mortalin and the complement membrane attack complexes are released in membrane vesicles from complement attacked cells. As shown here, knock down of mortalin with specific siRNA reduces MAC elimination and enhances cell sensitivity to MAC‐induced cell death. Similar results were obtained with MKT‐077, a cationic rhodacyanine dye that inhibits mortalin. Treatment of human erythroleukemia K562 and colorectal carcinoma HCT116 cells with MKT‐077 sensitizes them to cell death mediated by MAC but not by streptolysin O. Pre‐treatment of cells with MKT‐077 also reduces the extent of MAC‐mortalin vesiculation following a sublytic complement attack. In the presence of MKT‐077, the direct binding of mortalin to complement C9, the major MAC component, is inhibited. The tumor suppressor protein p53 is a known mortalin client protein. The effect of MKT‐077 on complement‐mediated lysis of HCT116 p53+/+ and p53−/− cells was found to be independent on the presence of p53. Our results also demonstrate that recombinant human mortain inhibits complement‐mediated hemolysis of rabbit erythrocytes as well as zinc‐induced C9 polymerization. We conclude that mortalin supports cancer cell resistance to complement‐dependent cytotoxicity and propose consideration of mortalin as a novel target for cancer adjuvant immunotherapy.
Gene‐expression‐based analysis of local and metastatic neuroblastoma variants reveals a set of genes associated with tumor progression in neuroblastoma patients
Authors: Nevo Ido , Oberthuer Andre , Botzer Edry , Sagi‐Assif Orit , Maman Shelly , Pasmanik‐Chor Metsada , Kariv Naam , Fischer Matthias , Yron Ilana , Witz Isaac P.
Published: September 2009

Metastasis is the primary cause of mortality in Neuroblastoma (NB) patients, but the metastatic process in NB is poorly understood. Metastsis is a multistep process that requires the coordinated action of many genes. The identification of genes that promote or suppress tumor metastasis can advance our understanding of this process. In the present study, we utilized a human NB xenograft model comprising local and metastatic NB variants, which was recently developed in our laboratory. We set out to identify molecular correlates of NB metastasis and to determine the clinical relevance of these molecules. We first performed genome‐wide expression profiles of metastatic and nonmetastatic NB variants that have an identical genetic background. We found that some of the proteins highly expressed in the metastatic NB variants are localized in the cytoplasm and endoplasmic reticulum. Other proteins are linked to metabolic processes and signaling pathways, thereby supporting the invasive and metastatic state of the cells. Subsequently, we intersected the differentially expressed genes in the human xenografted variants with genes differentially expressed in Stage 1 and Stage 4 primary tumors of NB patients. By using the same gene‐expression platform, molecular correlates associated with metastatic progression in primary NB tumors were identified. The resulting smaller gene set was clinically relevant as it discriminated between high‐ and low‐risk NB patients.
Sustained delivery and efficacy of polymeric nanoparticles containing osteopontin and bone sialoprotein antisenses in rats with breast cancer bone metastasis
Authors: Elazar Victoria , Adwan Hassan , Bäuerle Tobias , Rohekar Keren , Golomb Gershon , Berger Martin R.
Published: September 2009

Poor prognosis in mammary carcinoma is associated with a certain expression profile of a defined set of genes including osteopontin and bone sialoprotein. Efficient and specific delivery of antisenses (AS) and a protection of the sequences from degradation are the crucial conditions for AS therapeutic efficiency. We hypothesized that effective and safe AS delivery direceted against these genes could be achieved by polymeric nanoparticles (NP) fabricated from a biocompatible polymer. Due to their nano‐size range and small negative charge, AS‐NP can overcome the absorption barrier offering increased resistance to nuclease degradation, sustained duration of AS administration, and consequently, prolonged antisense action. The ASs designed against OPN and BSP‐II were successfully encapsulated in NP composed of the biodegradable and biocompatible polylactide‐co‐glycolide polymer (PLGA), exhibiting sustained release and stability of the ASs. The therapeutic efficacy of the AS‐NP delivery system was examined in vitro, and in a breast cancer bone metastasis animal model of MDA‐MB‐231 human breast cancer cells in nude rats. Treatment with OPN‐AS or BSP‐AS loaded NP in comparison with osmotic mini‐pumps (locoregional injection and SC implants, respectively) resulted in a significant decrease in both, tumor bone metastasis incidence and in the size of the lesions in rats with metastases. Despite its smaller dose, AS‐NP exhibited a better therapeutic efficacy than osmotic mini‐pumps in terms of lesion ratio at later time periods (8–12 weeks). It may be concluded that AS delivery by NP is a promising therapeutic modality providing stability of the encapsulated AS and a sustained release.
Comparison of HPV and cytology triage algorithms for women with borderline or mild dyskaryosis in population‐based cervical screening (VUSA‐screen study)
Authors: Rijkaart Dorien C. , Berkhof Johannes , van Kemenade Folkert J. , Rozendaal Lawrence , Verheijen René H.M. , Bulk Saskia , Herreilers Miriam E. , Verweij Wim M. , Snijders Peter J.F. , Meijer Chris J.L.M.
Published: September 2009

We studied the effectiveness of high‐risk human papillomavirus (hrHPV) triage for immediate colposcopy in women with borderline or mild dyskaryosis (BMD). In the Utrecht province of the Netherlands, women aged 30–60 years who participated in the regular cervical screening programme were offered hrHPV testing and cytology (intervention group) or cytology only (control group). In the intervention group (n = 337), women with BMD were immediately referred for colposcopy only if the sample was hrHPV positive. Women with a hrHPV negative test were advised to repeat cytology at 6 and 18 months and were referred for colposcopy if and when the repeat test result was positive (BMD or worse). In the control group (n = 329), referral of women with BMD was delayed until cytology was repeatedly positive at 6 or 18 months. The CIN3 detection rates were 10.7% (36/337) in the intervention group and 6.4% (21/329) in the control group (p = 0.047). Moreover, hrHPV triaging resulted in shorter time to diagnosis (154 vs. 381 days). Although the number of colposcopy referrals was 51.5% higher in the intervention group than in the control group, the medical costs per detected CIN3 were slightly lower ([euro] 4781 vs. [euro] 6235). If, in addition, hrHPV negative women had been referred back to routine screening at baseline, the CIN3 rate would have been 10.1% (34/337) and colposcopy rate would only have been 30.4% higher than in the control group. This study shows that hrHPV triaging of women with BMD is at least as effective for detecting CIN3 as repeat cytology, also when hrHPV negative women are referred back to routine screening.
Characterization of vascular structures and skin bruises using hyperspectral imaging, image analysis and diffusion theory
Authors: Randeberg Lise Lyngsnes , Larsen Eivind La Puebla , Svaasand Lars Othar
Published: September 2009

Hyperspectral imaging, image analysis and diffusion theory were used to visualize skin vasculature and to monitor the development of fresh skin bruises. Bruises were inflicted in a porcine model, and the development of the hemorrhage was monitored using white light hyperspectral imaging (400–1000 nm). Hyperspectral images from human volunteers were also included in the study. Statistical image analysis was used to classify bruised regions and to visualize the skin vasculature. Biopsies were collected from the animals to reveal the true depth of the bruising. A three‐layer diffusion model and an analytic hemoglobin transport model were used to model the reflectance spectra from the images. The results show that hyperspectral images contain depth information, and that the approximate depth and extent of bruises can be retrieved using a combination of statistical image analysis and diffusion theory. This technique also shows potential to visualize vascular structures in human skin. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
SUMF2 interacts with interleukin‐13 and inhibits interleukin‐13 secretion in bronchial smooth muscle cells
Authors: Liang Hongyan , Li Zhengjun , Xue Li , Jiang Xiaofeng , Liu Fang
Published: September 2009

IL‐13 is a central mediator of allergic inflammation and secreted by Th2 and bronchial smooth muscle cells (BSMC). However, little is known about the regulation of IL‐13 secretion. To address it, a cDNA library of BSMC was screened for the proteins interacted with IL‐13 by yeast two‐hybridization. Besides IL‐13 receptors, human sulfatase modifying factor 2 (SUMF2) was interacted with IL‐13. Furthermore, SUMF2 and IL‐13 were co‐immunoprecipitated from BSMC, which was independent of IL‐13 glycosylation. Interestingly, high levels of SUMF2 were expressed by BSMC, accompanied by significantly higher levels of intracellular IL‐13, but lower levels of IL‐13 secretion from BSMC. In contrast, little of SUMF2 was detected in lymphocytes, accompanied by lower levels of intracellular IL‐13, but significantly higher levels of 12 kDa form of IL‐13 secretion. Moreover, knockdown of SUMF2 expression by transfection with SUMF2‐specific siRNA did not alter IL‐13 mRNA transcription, but significantly reduced intracellular IL‐13 levels, associated with increased levels of IL‐13 secretion from BSMC. While induction of transient SUMF2 expression in lymphocytes failed to modulate IL‐13 mRNA transcription. It significantly increased the contents of 12 kDa form of intracellular IL‐13, accompanied by significantly reduced levels of IL‐13 in their supernatants. In addition, blockage of N‐glycosylation by treatment with tunicamycin eliminated 17 kDa form of intracellular IL‐13, but failed to promote IL‐13 secretion in BSMC. Collectively, our novel data clearly indicated that SUMF2 interacted with IL‐13 and inhibited IL‐13 secretion in BSMC and lymphocytes, which was independent of IL‐13 glycosylation. J. Cell. Biochem. 108: 1076–1083, 2009. © 2009 Wiley‐Liss, Inc.
Thrombin induces cyclooxygenase‐2 expression and prostaglandin E2 release via PAR1 activation and ERK1/2‐ and p38 MAPK‐dependent pathway in murine macrophages
Authors: Lo Huey‐Ming , Chen Chih‐Li , Tsai Yih‐Jeng , Wu Pi‐Hui , Wu Wen‐Bin
Published: September 2009

Thrombin levels increase at sites of vascular injury and during acute coronary syndromes. It is also increased several fold by sepsis with a reciprocal decrease in the anti‐thrombin III levels. In this study we investigate the effects of thrombin on the induction of cyclooxygenase‐2 (COX‐2) and prostaglandin (PG) production in macrophages. Thrombin‐induced COX‐2 protein and mRNA expression in RAW264.7 and primary cultured peritoneal macrophages. A serine proteinase, trypsin, also exerted a similar effect. The inducing effect by thrombin in macrophages was not affected by a lipopolysaccharide (LPS)‐binding antibiotic, polymyxin B, excluding the possibility of LPS contamination. The increase of COX‐2 expression by thrombin was functionally linked to release of PGE2 and PGI2 but not thromboxane A2 into macrophage culture medium. Thrombin‐induced COX‐2 expression and PGE2 production were significantly attenuated by PD98059 and SB202190 but not by SP600125, suggesting that ERK1/2 and p38 MAPK activation were involved in this process. This was supported by the observation that thrombin could directly activate ERK1/2 and p38 MAPK in macrophages. A further analysis indicated that the proteinase‐activated receptor 1 (PAR1)‐activating agonist induced effects similar to those induced by thrombin in macrophages and the PAR1 antagonist‐SCH79797 could attenuate thrombin‐induced COX‐2 expression and PGE2 release. Taken together, we provided evidence demonstrating that thrombin can induce COX‐2 mRNA and protein expression and PGE2 production in macrophages through PAR1 activation and ERK1/2 and p38 MAPK‐dependent pathway. The results presented here may explain, at least in part, the possible contribution of thrombin and macrophages in these pathological conditions. J. Cell. Biochem. 108: 1143–1152, 2009. © 2009 Wiley‐Liss, Inc.
Motivational interviewing: a bellwether for context‐responsive psychotherapy integration
Authors: Constantino Michael J. , DeGeorge Joan , Dadlani Mamta B. , Overtree Christopher E.
Published: September 2009

We comment on 6 clinical cases involving the application of one or more elements of Motivational interviewing (MI). First, we share our general reactions to MI and the case material. Second, we reflect briefly and specifically on each case illustration, highlighting the compelling flexibility and clinical utility of the MI spirit and its principles. Third, we offer several reflective themes across the cases, including convergences between MI and other psychotherapies, and unanswered clinical questions related to MI, its effectiveness, and its change mechanisms. Finally, we advance a context‐responsive psychotherapy integration for which MI might effectively serve as the bellwether. © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session 65: 1–8, 2009.
The effectiveness and applicability of motivational interviewing: a practice‐friendly review of four meta‐analyses
Authors: Lundahl Brad , Burke Brian L.
Published: September 2009

This article reviews the research support for Motivational interviewing (MI) so that practitioners can make informed decisions about the value and applicability of MI in their clinical work. We highlight the evidence from the three published meta‐analyses of MI and a recent meta‐analysis that we completed. MI is significantly (10%–20%) more effective than no treatment and generally equal to other viable treatments for a wide variety of problems ranging from substance use (alcohol, marijuana, tobacco, and other drugs) to reducing risky behaviors and increasing client engagement in treatment. Although most client‐related variables are unrelated to outcomes (e.g., age, gender, severity), some decisions about treatment format (e.g., individual vs. group) are important. For example, relying solely on group‐delivered MI appears to be less effective than one‐on‐one MI, whereas delivering MI with problem feedback is likely to generate better outcomes for some problems than MI alone. © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session 65: 1–14, 2009.
Motivation enhancement therapy can increase utilization of cognitive‐behavioral therapy: the case of social anxiety disorder
Authors: Buckner Julia D.
Published: September 2009

Despite the documented efficacy of cognitive‐behavioral therapy (CBT) for many psychological disorders, the vast majority of suffering people do not receive treatment. Treatment underutilization may prove at least partially due to ambivalence about seeking treatment. Ambivalence could result if, for example, people with social anxiety disorder (SAD) want to decrease their anxiety symptoms but are worried they will be judged negatively should they seek treatment. Motivation enhancement therapy (MET) can be a useful tool to help non‐treatment seekers explore and resolve ambivalence regarding seeking treatment. A case study illustrates the application of MET to encourage the utilization of CBT for a non‐treatment seeker with SAD. This case study includes outcome data regarding the client's experience with post‐MET CBT for her social anxiety. © 2009 Wiley Periodicals, Inc. J Clin Psychol: In Session 65: 1–12, 2009.
Regulation of intestinal stem cells in mammals and Drosophila
Authors: Wang Ping , Hou Steven X.
Published: September 2009

The digestive systems in mammals and Drosophila are quite different in terms of their complexity and organization, but their biological functions are similar. The Drosophila midgut is a functional equivalent of the mouse small intestine. Adult intestinal stem cells (ISCs) have been identified in both the mouse small intestine and Drosophila midgut. The anatomy and cell renewal in the Drosophila midgut are similar to those in the mouse small intestine: the intestinal epithelium in both systems is a tube composed of epithelial cells with absorptive and secretory functions; the Notch signaling controls absorptive versus secretory fate decisions in the intestinal epithelium; cell renewal in both systems starts from stem cells in the basal cell layer, and the differentiated cells then move toward the lumen. However, it is clear that the stem cells in the two systems are regulated in different ways. In this review, we will compare cell renewal and stem cell regulation in the two systems. J. Cell. Physiol. 222:33–37, 2010. © 2009 Wiley‐Liss, Inc.
Leptodactylus ocellatus (Amphibia): mechanism of defense in the skin and molecular phylogenetic relationships
Authors: Leite João Manoel Almeida , Silva Luciano Paulino , Silva‐Leite Roberta Rocha , Ferrari Ana Stella , Noronha Sergio Eustáquio , Silva Helio Ricardo , Bloch Carlos , Leite José Roberto de Souza de Almeida
Published: September 2009

Amphibian antimicrobial peptides have been known for many decades and several of them have already been isolated. However, the number of species investigated is still small. Herein, we report on the skin secretions of Leptodactylus ocellatus, which were extracted by mild electrical stimulation and its semi‐preparative reverse‐phase chromatography was resolved in more than 30 fractions. Among these fractions, two novel antimicrobial peptides were isolated and their amino acid sequences determined by de novo sequencing. The ocellatins‐5 and ‐6 (21 and 22 amino acid residues, respectively) are amidated at the C‐terminus. Ocellatins inhibited the growth of reference strains of both Gram‐negative bacteria (Escherichia coli) and Gram‐positive bacteria (Staphylococcus aureus) with minimal inhibition concentration values in the range of 32–128 µg/mL. The amino acid sequence of the peptides shows structural similarity with members of the antimicrobial peptides found in the skin secretion of other leptodactylid frogs. This observation is consistent with the hypothesis that many frog skin antimicrobial peptides are related evolutionarily, having arisen from multiple duplications of an ancestral gene that existed before the radiation of the different species. J. Exp. Zool. 313A:1–8, 2010. © 2009 Wiley‐Liss, Inc.
Evidence for urea‐induced hypometabolism in isolated organs of dormant ectotherms
Authors: Muir Timothy J. , Costanzo Jon P. , Lee Richard E.
Published: September 2009

Many organisms endure extended periods of dormancy by depressing their metabolism, which effectively prolongs the use of their endogenous energy stores. Though the mechanisms of hypometabolism are varied and incompletely understood, recent work suggests that urea accumulation in autumn and early winter contributes to reduced metabolism of hibernating wood frogs (Rana sylvatica). Urea accumulation during dormancy is a widespread phenomenon, and it has long been presumed that numerous species from diverse taxa benefit from its hypometabolic effect. To investigate the phylogenetic prevalence of urea‐induced hypometabolism, we studied four species of urea accumulators from the clades Amphibia (Spea bombifrons and Ambystoma tigrinum), Reptilia (Malaclemys terrapin), and Gastropoda (Anguispira alternata), and one amphibian species (R. pipiens) that does not accumulate urea during dormancy. We measured rates of oxygen consumption (V̇O2) of excised organ samples from dormant animals in the presence or absence of physiological concentrations of urea. Three of the four urea‐accumulating species had at least one organ whose V̇O2 was significantly decreased by urea treatment. However, V̇O2 of organs from R. pipiens, the one species tested that does not accumulate urea during dormancy, was not affected by urea treatment. Our results support the hypothesis that urea accumulation can reduce metabolic rate of dormant animals and provide a base for further investigation into the evolution of urea‐induced hypometabolism. J. Exp. Zool. 313A:28–34, 2010. © 2009 Wiley‐Liss, Inc.
Adult heat tolerance variation in Drosophila melanogaster is not related to Hsp70 expression
Authors: Jensen Louise Toft , Cockerell Fiona Elizabeth , Kristensen Torsten Nygaard , Rako Lea , Loeschcke Volker , McKechnie Stephen William , Hoffmann Ary Anthony
Published: September 2009

Expression of heat‐inducible Hsp70 is considered closely linked to thermotolerance in Drosophila melanogaster and other ectotherms. However, intra‐specific variation of Hsp70 expression levels and its relationship to heat resistance has only been investigated in a few studies. Although in Drosophila larvae Hsp70 expression may be a key determinant of heat tolerance, the evidence for this in adults is equivocal. We therefore examined heat‐induced Hsp70 expression and several measurements of adult heat tolerance in three independent collections of D. melanogaster, measured in three laboratories and using slightly different protocols. Expression levels of Hsp70 were quantified using ELISA or Western blots on extracts from adult females. Both Hsp70 and heat tolerance exhibited substantial within‐population variation as previously reported. However, in all experiments there were no significant correlation between Hsp70 expression and laboratory assays of adult heat tolerance commonly used in Drosophila. When combining data across three studies we had high power to detect associations but the results showed that variation in Hsp70 expression is only likely to explain a small proportion of variation in adult heat tolerance. Therefore, although Hsp70 expression is a major component of the cellular heat stress response, its influence on intra‐specific heat tolerance variation may be life‐stage specific. J. Exp. Zool. 313A:35–44, 2010. © 2009 Wiley‐Liss, Inc.
Directing human neural stem/precursor cells into oligodendrocytes by overexpression of Olig2 transcription factor
Authors: Maire Cécile L. , Buchet Delphine , Kerninon Christophe , Deboux Cyrille , Baron‐Van Evercooren Anne , Nait‐Oumesmar Brahim
Published: September 2009

Multipotential neural stem/precursor cells of the central nervous system were extensively studied for their properties of generating myelinating oligodendrocytes both in vitro and in vivo upon engraftment in animal models of myelin disorders, such as leucodystrophy and multiple sclerosis. These studies provided proof‐of‐principle that efficient myelination can be achieved by cell transplantation. However, one major drawback of cell‐based therapy of myelin diseases is the difficulty in generating oligodendrocytes efficiently from human fetal neural stem/precursor cells (hNPC). Here we explored whether overexpression of the basic helix‐loop‐helix (bHLH) transcription factor Olig2 in fetal hNPC could enhance the generation of oligodendrocytes both in vitro and in vivo. We report that transduction of hNPC with Olig2‐encoding lentiviral vectors enhances their commitment toward an oligodendroglial fate. Moreover, Olig2‐transduced hNPC, grafted into the dysmyelinated shiverer mouse brain, survived up to 9 weeks, migrated extensively, and differentiated into MBP+ myelinating oligodendrocytes. In contrast, control hNPC remained at a less mature stage and generated very few myelinating oligodendrocytes. Our study indicates that bHLH transcription factors, such as Olig2, are interesting targets for directing hNPC into myelinating oligodendrocytes. © 2009 Wiley‐Liss, Inc.
Microglia of rat ventral midbrain recovers its resting state over time in vitro: Let microglia rest before work
Authors: Cristóvão Ana Clara , Saavedra Ana , Fonseca Carla Pais , Campos Filipa , Duarte Emília Pedrosa , Baltazar Graça
Published: September 2009

Cortical or total brain cultures of microglia are commonly used as a model to study the inflammatory processes in Parkinson's disease. Here we characterize microglia cultures from rat ventral midbrain and evaluate their response to zymosan A. We used specific markers of microglia and evaluated the morphology, the phagocytic activity and reactive oxygen species (ROS) levels of the cells. During the first 10 days in vitro (DIV), cultures presented predominantly cells with a round morphology, expressing CD68 and with high phagocytic activity and ROS production. After 13 DIV, this tendency was reversed, with cultures showing higher number of ramified cells and fewer CD68+ cells along with lower phagocytic and ROS production capability, suggesting that microglia must be kept in vitro for at least 13 days to recover its resting state. The exposure of cultures with less than 10 DIV to zymosan A significantly decreased cell viability. Exposure of cultures with 13 DIV to zymosan A (0.05, 0.5, or 5 μg/ml) increased the total cell number, the percentage of CD68+ cells, and the phagocytic activity. Concentrations of zymosan A higher than 5 μg/ml were also effective in activating microglia but significantly decreased the number of viable cells. In summary, microglial cells remain in the activated state for several days after the isolation process and, thus, stimulation of microglia recently isolated can compromise interpretation of the results. However, upon 13 DIV, cells achieve properties of nonactivated microglia and present a characteristic response to a proinflammatory agent. © 2009 Wiley‐Liss, Inc.
Up‐regulation of oligodendrocyte precursor cell αV integrin and its extracellular ligands during central nervous system remyelination
Authors: Zhao Chao , Fancy Stephen P.J. , Franklin Robin J.M. , ffrench‐Constant Charles
Published: September 2009

To determine the role of extracellular matrix molecules and their integrin ligands in CNS remyelination, we have examined in experimentally induced focal demyelinated lesions the expression of the two classes of integrins implicated in oligodendrocyte development and myelination: α6 laminin‐binding integrins and αV integrins that bind a range of extracellular matrix proteins containing the ‐Arg‐Gly‐Asp‐ (RGD) recognition sequence. Only αV integrins were up‐regulated during remyelination, being expressed on oligodendrocyte precursor cells during their recruitment into the lesion. Next, therefore, we examined the expression of extracellular matrix ligands for αV integrins and documented increased expression of tenascin‐C, tenascin‐R, fibronectin, and vitronectin. Taken together with our previous discovery of high levels of expression of another αV ligand, osteopontin, during remyelination in these lesions, our findings suggest that αV integrins make an important contribution to successful repair in the CNS. © 2009 Wiley‐Liss, Inc.
Lack of aspartoacylase activity disrupts survival and differentiation of neural progenitors and oligodendrocytes in a mouse model of Canavan disease
Authors: Kumar Shalini , Biancotti Juan Carlos , Matalon Reuben , de Vellis Jean
Published: September 2009

Loss of the oligodendrocyte (OL)‐specific enzyme aspartoacylase (ASPA) from gene mutation results in the sponginess and loss of white matter (WM) in Canavan disease (CD). This study addresses the fate of OLs during the pathophysiology of CD in an adult ASPA knockout (KO) mouse strain. Massive arrays of neural stem/progenitor cells, immunopositive for PSA‐NCAM, nestin, vimentin, and NG2, were observed within the severely affected spongy WM of the KO mouse brain. In these mice, G1→S cell cycle progression was confirmed by an increase in cdk2‐kinase activity, a reduction in mitotic inhibitors p21Cip1 and p27Kip1, and an increase in bromodeoxyuridine (BrdU) incorporation. Highly acetylated nuclear histones H2B and H3 were detected in adult KO mouse WM, suggesting the existence of noncompact chromatin as seen during early development. Costaining for BrdU‐ or Ki67‐positive cells with markers for neural progenitors confirmed a continuous generation of OL lineage cells in KO WM. We observed a severe reduction in 21.5‐ and 18.5‐kDa myelin basic protein and PLP/DM20 proteolipid proteins combined with a decrease in myelinated fibers and a perinuclear retention of myelin protein staining, indicating impairment in protein trafficking. Death of OLs, neurons, and astrocytes was identified in every region of the KO brain. Immature OLs constituted the largest population of dying cells, particularly in WM. We also report an early expression of full‐length ASPA mRNA in normal mouse brain at embryonic day 12.5, when OL progenitors first appear during development. These findings support involvement of ASPA in CNS development and function. © 2009 Wiley‐Liss, Inc.