bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of the Aragonitic Layer in Adult Oyster Shells, Crassostrea gigas: Structural Study of Myostracum including the Adductor Muscle Scar
Authors: Lee Seung-Woo , Jang Young-Nam , Kim Jeong-Chan
Published: June 2011

Myostracum, which is connected from the umbo to the edge of a scar, is not a single layer composed of prismatic layers, but a hierarchically complex multilayered shape composed of minerals and an organic matrix. Through the analysis of the secondary structure, the results revealed that a β‐antiparallel structure was predominant in the mineral phase interface between the myostracum (aragonite) and bottom folia (calcite). After the complete decalcification and deproteinization, the membrane obtained from the interface between the myostracum buried in upper folia, and the bottom folia was identified as chitin. The transitional zone in the interface between the adductor muscle scar and folia are verified. The myostracum disappeared at the edge of the scar of the posterior side. From this study, the entire structure of the myostracum from the adult oyster shell of Crassostrea gigas could be proposed.
Effect of a Semisolid Formulation ofLinum usitatissimumL. (Linseed) Oil on the Repair of Skin Wounds
Authors: Franco Eryvelton de Souza , de Aquino Camilla Maria Ferreira , Medeiros Paloma Lys de , Evêncio Liriane Baratella , Góes Alexandre José da Silva , Maia Maria Bernadete de Souza
Published: June 2011

The purpose of this study was to investigate the effects of a semisolid formulation of linseed oil, SSFLO (1%, 5%, or 10%) orin naturalinseed oil on skin wounds of rats. We used wound models, incisional and excisional, to evaluate, respectively, the contraction/reepithelialization of the wound and resistance to mechanical traction. The groups (n=6) treated with SSFLO (1% or 5%) began the process of reepithelialization, to a significant extent (P<.05), on the sixth day, when compared to the petroleum jelly control group. On 14th day for the groups treated with SSFLO (1% or 5%), 100% reepithelialization was found, while in the petroleum jelly control group, this was only 33.33%. Our study showed that topical administration of SSFLO (1% or 5%) in excisional wounds allowed reepithelialization in 100% of treated animals. Therefore, a therapeutic potential of linseed oil, when used at low concentrations in the solid pharmaceutical formulations, is suggested for the process of dermal repair.
Surgical Castration in Hormone-Refractory Metastatic Prostate Cancer Patients Can Be an Alternative for Medical Castration
Authors: Zaitsu Masayoshi , Yamanoi Mariko , Mikami Koji , Takeshima Yuta , Okamoto Naohiko , Imao Sadao , Tonooka Akiko , Takeuchi Takumi
Published: June 2011

Background. Most patients with metastatic prostate cancer are endocrinologically treated with LHRH agonist, but finally castration-refractory and hormone-refractory cancers occur. Serum testosterone levels get low to “the castration level” by LHRH agonists but may not get low enough against castration-refractory prostate cancer.Methods. As case series, twelve patients suffering from hormone-refractory prostate cancer continuously on LHRH agonist underwent surgical castration. Additionally, one hundred and thirty-nine prostate cancer patients on LHRH agonist or surgical castration were tested for serum total testosterone levels.Results. Surgical castration caused decrease in serum PSA in one out of 12 hormone-refractory prostate cancer patients with PSA reduction rate 74%. Serum total testosterone levels were below the sensitivity threshold (0.05 ng/mL) in 40 of 89 (44.9%) medically castrated patients and 33 of 50 (66.0%) surgically castrated patients (P=.20).Conclusion. Even hormone-refractory prostate cancer patients are candidates for surgical castration because of endocrinological, oncological, and economical reasons.
A Low Carbohydrate, High Protein Diet Slows Tumor Growth and Prevents Cancer Initiation
Authors: Ho Victor W. , Leung Kelvin , Hsu Anderson , Luk Beryl , Lai June , Shen Sung Yuan , Minchinton Andrew I. , Waterhouse Dawn , Bally Marcel B. , Lin Wendy , Nelson Brad H. , Sly Laura M. , Krystal Gerald
Published: June 2011

Since cancer cells depend on glucose more than normal cells, we compared the effects of low carbohydrate (CHO) diets to a Western diet on the growth rate of tumors in mice. To avoid caloric restriction–induced effects, we designed the low CHO diets isocaloric with the Western diet by increasing protein rather than fat levels because of the reported tumor-promoting effects of high fat and the immune-stimulating effects of high protein. We found that both murine and human carcinomas grew slower in mice on diets containing low amylose CHO and high protein compared with a Western diet characterized by relatively high CHO and low protein. There was no weight difference between the tumor-bearing mice on the low CHO or Western diets.

Additionally, the low CHO-fed mice exhibited lower blood glucose, insulin, and lactate levels. Additive antitumor effects with the low CHO diets were observed with the mTOR inhibitor CCI-779 and especially with the COX-2 inhibitor Celebrex, a potent anti-inflammatory drug. Strikingly, in a genetically engineered mouse model of HER-2/neu–induced mammary cancer, tumor penetrance in mice on a Western diet was nearly 50% by the age of 1 year whereas no tumors were detected in mice on the low CHO diet. This difference was associated with weight gains in mice on the Western diet not observed in mice on the low CHO diet. Moreover, whereas only 1 mouse on the Western diet achieved a normal life span, due to cancer-associated deaths, more than 50% of the mice on the low CHO diet reached or exceeded the normal life span. Taken together, our findings offer a compelling preclinical illustration of the ability of a low CHO diet in not only restricting weight gain but also cancer development and progression. Cancer Res; 71(13); 4484–93. ©2011 AACR.
BMP4 Promotes Prostate Tumor Growth in Bone through Osteogenesis
Authors: Lee Yu-Chen , Cheng Chien-Jui , Bilen Mehmet A. , Lu Jing-Fang , Satcher Robert L. , Yu-Lee Li-Yuan , Gallick Gary E. , Maity Sankar N. , Lin Sue-Hwa
Published: June 2011

Induction of new bone formation is frequently seen in the bone lesions from prostate cancer. However, whether osteogenesis is necessary for prostate tumor growth in bone is unknown. Recently, 2 xenografts, MDA-PCa-118b and MDA-PCa-133, were generated from prostate cancer bone metastases. When implanted subcutaneously in severe combined immunodeficient (SCID) mice, MDA-PCa-118b induced strong ectopic bone formation while MDA-PCa-133 did not. To identify the factors that are involved in bone formation, we compared the expression of secreted factors (secretome) from MDA-PCa-118b and MDA-PCa-133 by cytokine array. We found that the osteogenic MDA-PCa-118b xenograft expressed higher levels of bone morphogenetic protein BMP4 and several cytokines including interleukin-8, growth-related protein (GRO), and CCL2. We showed that BMP4 secreted from MDA-PCa-118b contributed to about a third of the osteogenic differentiation seen in MDA-PCa-118b tumors. The conditioned media from MDA-PCa-118b induced a higher level of osteoblast differentiation, which was significantly reduced by treatment with BMP4 neutralizing antibody or the small molecule BMP receptor 1 inhibitor LDN-193189. BMP4 did not elicit an autocrine effect on MDA-PCa-118b, which expressed low to undetectable levels of BMP receptors. Treatment of SCID mice bearing MDA-PCa-118b tumors with LDN-193189 significantly reduced tumor growth. Thus, these studies support a role of BMP4-mediated osteogenesis in the progression of prostate cancer in bone. Cancer Res; 71(15); 5194–203. ©2011 AACR.
Microvesicles Released from Human Renal Cancer Stem Cells Stimulate Angiogenesis and Formation of Lung Premetastatic Niche
Authors: Grange Cristina , Tapparo Marta , Collino Federica , Vitillo Loriana , Damasco Christian , Deregibus Maria Chiara , Tetta Ciro , Bussolati Benedetta , Camussi Giovanni
Published: June 2011

Recent studies suggest that tumor-derived microvesicles (MV) act as a vehicle for exchange of genetic information between tumor and stromal cells, engendering a favorable microenvironment for cancer development. Within the tumor mass, all cell types may contribute to MV shedding, but specific contributions to tumor progression have yet to be established. Here we report that a subset of tumor-initiating cells expressing the mesenchymal stem cell marker CD105 in human renal cell carcinoma releases MVs that trigger angiogenesis and promote the formation of a premetastatic niche. MVs derived only from CD105-positive cancer stem cells conferred an activated angiogenic phenotype to normal human endothelial cells, stimulating their growth and vessel formation after in vivo implantation in immunocompromised severe combined immunodeficient (SCID) mice. Furthermore, treating SCID mice with MVs shed from CD105-positive cells greatly enhanced lung metastases induced by i.v. injection of renal carcinoma cells. Molecular characterization of CD105-positive MVs defines a set of proangiogenic mRNAs and microRNAs implicated in tumor progression and metastases. Our results define a specific source of cancer stem cell–derived MVs that contribute to triggering the angiogenic switch and coordinating metastatic diffusion during tumor progression. Cancer Res; 71(15); 5346–56. ©2011 AACR.
ERK1/2 and p38α/β Signaling in Tumor Cell Quiescence: Opportunities to Control Dormant Residual Disease
Authors: Sosa Maria Soledad , Avivar-Valderas Alvaro , Bragado Paloma , Wen Huei-Chi , Aguirre-Ghiso Julio A.
Published: June 2011

Systemic minimal residual disease after primary tumor treatment can remain asymptomatic for decades. This is thought to be due to the presence of dormant disseminated tumor cells (DTC) or micrometastases in different organs. DTCs lodged in brain, lungs, livers, and/or bone are a major clinical problem because they are the founders of metastasis, which ultimately kill cancer patients. The problem is further aggravated by our lack of understanding of DTC biology. In consequence, there are almost no rational therapies to prevent dormant DTCs from surviving and expanding. Several cancers, including melanoma as well as breast, prostate, and colorectal carcinomas, undergo dormant periods before metastatic recurrences develop. Here we review our experience in studying the cross-talk between ERK1/2 and p38α/β signaling in models of early cancer progression, dissemination, and DTC dormancy. We also provide some potential translational and clinical applications of these findings and describe how some currently used therapies might be useful to control dormant disease. Finally, we draw caution on the use of p38 inhibitors currently in clinical trials for different diseases as these may accelerate metastasis development. Clin Cancer Res; 17(18); 5850–7. ©2011 AACR.
Fibroblast Growth Factor Receptors Are Components of Autocrine Signaling Networks in Head and Neck Squamous Cell Carcinoma Cells
Authors: Marshall Marianne E. , Hinz Trista K. , Kono Scott A. , Singleton Katherine R. , Bichon Brady , Ware Kathryn E. , Marek Lindsay , Frederick Barbara A. , Raben David , Heasley Lynn E.
Published: June 2011

Purpose: We previously reported that a fibroblast growth factor (FGF) receptor (FGFR) signaling pathway drives growth of lung cancer cell lines of squamous and large cell histologies. Herein, we explored FGFR dependency in cell lines derived from the tobacco-related malignancy, head and neck squamous cell carcinoma (HNSCC).

Experimental Design: FGF and FGFR mRNA and protein expression was assessed in nine HNSCC cell lines. Dependence on secreted FGF2 for cell growth was tested with FP-1039, an FGFR1-Fc fusion protein. FGFR and epidermal growth factor receptor (EGFR) dependence was defined by sensitivity to multiple inhibitors selective for FGFRs or EGFR.

Results: FGF2 was expressed in eight of the nine HNSCC cell lines examined. Also, FGFR2 and FGFR3 were frequently expressed, whereas only two lines expressed FGFR1. FP-1039 inhibited growth of HNSCC cell lines expressing FGF2, identifying FGF2 as an autocrine growth factor. FGFR inhibitors selectively reduced in vitro growth and extracellular signal-regulated kinase signaling in three HNSCC cell lines, whereas three distinct lines exhibited responsiveness to both EGFR and FGFR inhibitors. Combinations of these drugs yielded additive growth inhibition. Finally, three cell lines were highly sensitive to EGFR tyrosine kinase inhibitors (TKI) with no contribution from FGFR pathways.

Conclusions: FGFR signaling was dominant or codominant with EGFR in six HNSCC lines, whereas three lines exhibited little or no role for FGFRs and were highly EGFR dependent. Thus, the HNSCC cell lines can be divided into subsets defined by sensitivity to EGFR and FGFR-specific TKIs. FGFR inhibitors may represent novel therapeutics to deploy alone or in combination with EGFR inhibitors in HNSCC. Clin Cancer Res; 17(15); 5016–25. ©2011 AACR.
Why Is This Effective HSP90 Inhibitor Not Being Developed in HER2+ Breast Cancer?
Authors: Arteaga Carlos L.
Published: June 2011

Inhibition of the HSP90 chaperone leads to degradation of the HER2 receptor. The HSP90 inhibitor tanespimycin in combination with trastuzumab is active in patients with HER2-overexpressing metastatic breast cancer. This combination is one of several HER2-targeted therapies that will significantly improve the outcome of patients with this subtype of breast cancer. Clin Cancer Res; 17(15); 4919–21. ©2011 AACR.
Neutralizing Monoclonal Antibody to Periostin Inhibits Ovarian Tumor Growth and Metastasis
Authors: Zhu Min , Saxton Romaine E. , Ramos Lillian , Chang David D. , Karlan Beth Y. , Gasson Judith C. , Slamon Dennis J.
Published: June 2011

Periostin, an extracellular matrix protein, is reported to be overexpressed in a variety of human cancers and its functions seem to be linked to tumor metastasis. Our previous results show that engineered periostin overexpression promotes ovarian tumor growth and dissemination in vivo. In this study, we developed a neutra-lizing monoclonal antibody to periostin, named MZ-1, and investigated its effects on human ovarian tumor growth and metastasis. Our in vivo studies showed significant growth inhibition by MZ-1 on both subcutaneous and intraperitoneal (i.p.) tumors derived from the periostin-expressing ovarian cancer cell line A2780. In addition, MZ-1 treatment led to a reduction of the metastatic potential of these A2780 i.p. tumors. The in vivo antitumor effects of MZ-1 were linked to its specific inhibition of anchorage-independent growth and survival of periostin-expressing cells, as well as its neutralizing effects on periostin-induced cancer cell migration and invasion. The data suggest that blocking periostin expression may be a novel approach for treating the subset of invasive ovarian tumors that overexpress periostin protein. Mol Cancer Ther; 10(8); 1500–8. ©2011 AACR.
A Pilot Clinical Study of Treatment Guided by Personalized Tumorgrafts in Patients with Advanced Cancer
Authors: Hidalgo Manuel , Bruckheimer Elizabeth , Rajeshkumar N.V. , Garrido-Laguna Ignacio , De Oliveira Elizabeth , Rubio-Viqueira Belen , Strawn Steven , Wick Michael J. , Martell James , Sidransky David
Published: June 2011

Patients with many advanced solid cancers have very poor prognosis, and improvements in life expectancy are measured only in months. We have recently reported the remarkable clinical outcome of a patient with advanced, gemcitabine-resistant, pancreatic cancer who was later treated with DNA-damaging agents, on the basis of the observation of significant activity of this class of drugs against a personalized tumorgraft generated from the patient's surgically resected tumor. Here, we extend the approach to patients with other advanced cancers. Tumors resected from 14 patients with refractory advanced cancers were propagated in immunodeficient mice and treated with 63 drugs in 232 treatment regimens. An effective treatment regimen in the xenograft model was identified for 12 patients. One patient died before receiving treatment, and the remaining 11 patients received 17 prospectively guided treatments. Fifteen of these treatments resulted in durable partial remissions. In 2 subjects, no effective treatments were found. Overall, there was a remarkable correlation between drug activity in the model and clinical outcome, both in terms of resistance and sensitivity. The data support the use of the personalized tumorgraft model as a powerful investigational platform for therapeutic decision making and to efficiently guide cancer treatment in the clinic. Mol Cancer Ther; 10(8); 1311–6. ©2011 AACR.
Is Serum Human Chorionic Gonadotrophin Follow-Up Necessary after Suspected Spillage of Trophoblast at the Time of Laparoscopic Surgery for Ectopic Pregnancy?
Authors: Bora Shabana A. , Kirk Emma , Daemen Anneleen , Timmerman Dirk , Bourne Tom
Published: June 2011

Background: Persistent trophoblast is a recognised complication of salpingostomy for the treatment of ectopic pregnancy, with reported rates of 3–20%; hence, women are advised to have serum human chorionic gonadotrophin (hCG) levels monitored post-operatively. Although much less common, there are also reports of disseminated trophoblastic peritoneal implants after laparoscopic salpingectomy. The aim of this study was to assess whether monitoring of post-operative serum hCG levels is necessary in women undergoing salpingectomy, where intra-operative spillage of trophoblast is thought to have occurred. Methods: This was a retrospective study of women who underwent serum hCG follow-up after salpingectomy. Serum hCG levels were monitored if: (1) the ectopic pregnancy was found to be ruptured; (2) there was a significant haemoperitoneum (>500 ml); (3) there was thought to be spillage of trophoblast at the time of salpingectomy or (4) a tubal miscarriage was diagnosed. Serum hCG levels were taken at days 1–2, days 3–4, days 6–8 or days 13–15 post-surgery. Women were followed up until the serum hCG level was <15 IU/l. Persistent trophoblast was defined as a failure of the serum hCG level to decrease spontaneously after surgery. Results: 105 women underwent serum hCG follow-up after a laparoscopy for a tubal ectopic pregnancy. Of these women, 92 had a laparoscopic salpingectomy and 13 were diagnosed with a tubal miscarriage at the time of laparoscopy. In all women the serum hCG decreased spontaneously. Conclusion: It does not appear necessary to routinely monitor serum hCG levels post-operatively in women diagnosed with tubal miscarriages, in those undergoing salpingectomy for a ruptured ectopic pregnancy or in cases of salpingectomy, where there is thought to be spillage of trophoblast.
Comparison of Long-Term Outcome between Doublet and Triplet Neoadjuvant Chemotherapy in Non-Metastatic Osteosarcoma of the Extremity
Authors: Hong Soojung , Shin Sang Joon , Jung Minkyu , Jeong Jaeheon , Lee Young Joo , Shin Kyoo-Ho , Roh Jae Kyung , Rha Sun Young
Published: June 2011

Objective: This study compared outcomes between doublet (AP) and triplet (IAP) neoadjuvant chemotherapy for nonmetastatic osteosarcoma of the extremity. Methods: A total of 124 patients were enrolled. In the AP group, a doublet regimen of intraarterial cisplatin and intravenous doxorubicin was given to 77 patients from 1991 to 1999. In the IAP group, a triplet regimen of additional intravenous ifosfamide was given to 47 patients from 2000 to 2007. After completion of 3 cycles of chemotherapy, patients underwent surgery. We assessed tumor response according to pathologic tumor necrosis, and treated patients with further adjuvant chemotherapy. Results: The overall pathologic response was excellent with more than 90% tumor necrosis in 74.8% of patients. Total necrosis of tumors was also found in 46 (37.4%) patients. There was no difference between the 2 groups in pathologic response (75.3 vs. 72.3%; p = 0.52) or other clinicopathologic parameters. There was no difference between the 2 groups in recurrence rate (31.2 vs. 31.9%; p = 0.17) or lung metastasis (28.6 vs. 23.4%; p = 0.53). Moreover, there were no statistical differences in median disease-free survival and overall survival between the groups. There was more hematologic toxicity in the IAP group (neutropenia, p = 0.002; thrombocytopenia, p = 0.001; febrile neutropenia, p < 0.001). Conclusions: The addition of ifosfamide to doxorubicin and cisplatin in neoadjuvant chemotherapy did not show improved outcomes in this study. Further trials are required to elucidate optimal neoadjuvant chemotherapy and effective salvage regimens.
The Honeybee Antimicrobial Peptide Apidaecin Differentially Immunomodulates Human Macrophages, Monocytes and Dendritic Cells
Authors: Tavano Regina , Segat Daniela , Gobbo Marina , Papini Emanuele
Published: June 2011

We show that apidaecin binds to human macrophages, monocytes and dendritic cells, displaying different intracellular distributions and inducing diversified effects. An apidaecin-cell association was detectable at concentrations as low as 5 µM and increased without saturation until 60 µM, was receptor independent and required a physiological temperature (37°C). For apidaecin, cytosolic localization was prevalent in macrophages and endosomal localization in monocytes, and associations with the plasma membrane were predominant in dendritic cells. Apidaecin upregulated T-lymphocyte co-stimulatory molecule CD80 and cytokine/chemokine production in macrophages, but not in monocytes and dendritic cells. Suboptimal stimulatory doses (5–10 µM) of apidaecin partially inhibited the lipopolysaccharide (LPS)-induced increase in major histocompatibility complex class II (MHCII) and CD86 in macrophages, and the release of selected cytokines/chemokines by both macrophages [interleukin (IL)-6 and tumor necrosis factor (TNF)-α] and monocytes [IL-6, TNF-α, basic fibroblast growth factor (FGF) and eotaxin]. Apidaecin had a double-edged effect: at low concentrations it partially antagonized LPS-stimulatory effects on both macrophages and monocytes while it stimulated pro-inflammatory and pro-immune functions of macrophages at higher concentrations.
A Prognostic Model to Predict Clinical Outcome in Gastric Cancer Patients with Bone Metastasis
Authors: Park Hyung Soon , Rha Sun Young , Kim Hyo Song , Hyung Woo Jin , Park Ji Soo , Chung Hyun Cheol , Noh Sung Hoon , Jeung Hei-Cheul
Published: June 2011

Background: The clinicopathological manifestations and treatment outcomes of bone metastasis of gastric cancer are largely unknown. We delineated a prognostic model to identify different risk groups on the basis of clinical parameters. Methods: Patients who had bone metastasis at the time of diagnosis of gastric cancer (synchronous metastasis) or who developed bone metastasis during follow-up (metachronous metastasis) were retrospectively reviewed from January 1998 to May 2008. Results: Bone metastasis was identified in 203 (2.4%) of 8,633 patients: 126 patients (62%) with synchronous metastasis and 77 patients with metachronous metastasis. The median time to event was 16 months (range 4–87). As for treatment, 120 patients (59%) received systemic chemotherapy. The median survival time was 103 days (95% CI 80–126). Poor performance status [Eastern Cooperative Oncology Group 3–4; relative risk (RR) = 1.91, p = 0.011], multiple bone metastasis (RR = 2.593, p = 0.002), and abnormal carcinoembryonic antigen (RR = 1.779, p = 0.004) implied independent factors for survival. For patients who had zero to two of these factors identified, chemotherapy had a beneficial effect (175 vs. 43 days; p < 0.0001). Conclusion: We recommend that the therapeutic approach with bone metastasis be customized to facilitate the risk stratification, so as to consequently provide the most appropriate therapy for each patient.
Time-Course Studies of Implanted Rabbit VX2 Liver Tumors to Identify the Appropriate Time for Starting Hepatic Arterial Embolization in Animal Models
Authors: Sonoda Akinaga , Nitta Norihisa , Nitta-Seko Ayumi , Ohta Shinichi , Nagatani Yukihiro , Mukaisho Kenichi , Otani Hideji , Tsuchiya Keiko , Takahashi Masashi , Murata Kiyoshi
Published: June 2011

Purpose: We followed the 4-week course of implanted VX2 tumors in rabbits and compared MRI and pathological findings to determine the appropriate time for starting therapy in animal liver tumor models. Materials and Methods: We used 18 Japanese white rabbits. The VX2 liver tumor was harvested from one tumor-bearing rabbit and implanted in the liver of the other 17 rabbits. They were then sacrificed at 1 (n = 5), 2 (n = 3), 3 (n = 4), and 4 weeks (n = 5) after implantation and MRI study. Using MRI scans and/or pathological specimens of individual rabbits, we evaluated the tumor survival ratio, the major tumor axes, intrahepatic metastases, and peritoneal dissemination. Results: All tumor transplantations were successful. At 1 week, 56.25% of the implanted tumors were visualized on MRI scans. At 2 weeks or later, all transplanted rabbits were confirmed to be tumor-bearing on MRI scans. At 3 weeks after implantation, the tumor size was similar on MRI scans and in pathological specimens. There were no intra-hepatic metastases or peritoneal disseminations within 2 weeks of tumor transplantation. Conclusion: We suggest that in studies of implanted VX2 models addressing the treatment of solid hepatic tumors, it may be prudent to start hepatic arterial embolization at 2 weeks after implantation.
Therapeutic Outcomes of Papillary Thyroid Cancer Patients in Different Risk Groups
Authors: Huang Bie-Yu , Lin Jen-Der , Chao Tzu-Chieh , Lin Kun-Ju , Hseuh Chuen , Tsang Ngan-Ming
Published: June 2011

Objective: The objective of this study was to determine the therapeutic outcome of papillary thyroid cancer (PTC) patients in different risk groups in one institute. Methods: A total of 1,759 PTC patients were categorized into low- (n = 1,123), intermediate- (n = 75), and high-risk (n = 561) groups according to tumor-node-metastasis (TNM) stage. Results: Of the patients, 15.1% presented with lymph node metastases, and 4.6% presented with distant metastases at the time of thyroid operation. After 8.0 ± 0.1 years of follow-up, 73 (4.2%) patients died of thyroid cancer. Tumor size, local invasion, and lymph node metastases adversely influenced recurrence and survival. Of the patients in the 3 groups, 9 (0.8%), 8 (10.7%), and 56 (10.0%) died of thyroid cancer, respectively. In addition, 88 (7.8%), 14 (18.7%), and 144 (25.8%) patients showed recurrence during the follow-up period. Patients with highly aggressive histological patterns showed increased recurrence and cancer mortality compared with the low-risk group; otherwise, values were not higher than those of the high-risk group. Conclusions: The cancer-related mortality was nearly 10% in the intermediate- and high-risk groups, and the patients in these groups required aggressive surgical and postoperative adjuvant therapies.
Prognostic Impact of Phosphorylated Mitogen-Activated Protein Kinase Expression in Patients with Metastatic Gastric Cancer
Authors: Atmaca Akin , Pauligk Claudia , Steinmetz Kristina , Altmannsberger Hans-Michael , Jäger Elke , Al-Batran Salah-Eddin
Published: June 2011

Background: This study was designed to investigate the expression of phosphorylated mitogen-activated protein kinase (p-MAPK/Erk1 and Erk2) and its correlation with outcomes in patients with metastatic gastric cancer. Methods: p-MAPK was detected by immunohistochemistry using monoclonal antibodies in a total of 223 formalin-fixed, paraffin-embedded samples obtained from 156 patients who received first-line chemotherapy in a phase III trial. Results: p-MAPK was positive in 93 (59.6%) and negative in 63 (40.4%) of the 156 patients evaluated. Similar rates of p-MAPK positivity were found in primaries (53%) and metastatic lesions (61.4%). Overall survival was significantly shorter in p-MAPK-positive patients (13.7 vs. 8.5 months) in the univariate analysis. However, this prognostic value disappeared as a trend in the multivariate analysis (p = 0.1). There was a strong, positive correlation between p-MAPK and the MIB-1 proliferation index, but MIB-1 did not predict outcomes. Conclusion: p-MAPK expression could be a potential negative prognostic parameter in patients with metastatic gastric cancer treated with chemotherapy.
Phosphate-Induced Apoptosis in Human Peritoneal Mesothelial Cells in vitro
Authors: Park Jong-Won , Yook Ju-Min , Ryu Hye-Myung , Choi Soon-Youn , Morishita Masayo , Do Jun-Young , Park Sun-Hee , Kim Chan-Duck , Choi Ji-Young , Chung Ho-Young , Kim Yong-Lim
Published: June 2011

Background: It has been demonstrated that phosphate uptake through the type III sodium-dependent phosphate co-transporter, Pit-1, induced apoptosis of aortic vascular smooth muscle cells and endothelial cells in vitro. However, the apoptotic effects of high phosphate (HP) level in human peritoneal mesothelial cells (HPMCs) are not known. Methods: To examine whether Pit-1 is expressed in HPMCs, we checked the Western blot assay of immunoreactive Pit-1 and the transcription of Pit-1 by reverse transcriptase PCR. We treated several different phosphate concentrations (1–4 mM) and calcium concentrations (1.8 and 2.8 mM) on HPMCs to assess the effects of concentration. MTT, TUNEL assays, and flow cytometry analysis using Annexin V and propidium iodide were performed to identify cell death and apoptosis. Bax and Bcl-2 by Western blot and caspase-3 activity were evaluated by colorimetric assay. In addition, phosphonoformic acid (PFA) and pan-caspase inhibitor, Z-VAD-FMK, were given to prevent phosphate-induced apoptosis. Results: Pit-1 expression on HPMCs was demonstrated. Apoptosis in HPMCs significantly increased with a high concentration of phosphate in a dose- and time-dependent manner, and was enhanced in the presence of 2.8 mM calcium. HP concentrations significantly decreased the anti-apoptotic Bcl-2/Bax ratio and increased caspase-3 activity. The treatment with PFA and Z-VAD-FMK prevented cell death by HP. Conclusion: Phosphate uptake through Pit-1 induces apoptosis in HPMCs by a caspase-related mechanism.
Smooth Muscle Cell Differentiation In Vitro
Authors: Xie Changqing , Ritchie Raquel P. , Huang Huarong , Zhang Jifeng , Chen Y. Eugene
Published: June 2011

Development of in vitro models by which to study smooth muscle cell (SMC) differentiation has been hindered by some peculiarities intrinsic to these cells, namely their different embryological origins and their ability to undergo phenotypic modulation in cell culture. Although many in vitro models are available for studying SMC differentiation, careful consideration should be taken so that the model chosen fits the questions being posed. In this review, we summarize several well-established in vitro models available to study SMC differentiation from stem cells and outline novel mechanisms recently identified as underlying SMC differentiation programs.
The Spatial and Developmental Relationships in the Macrophage Family
Authors: Swirski Filip K.
Published: June 2011

A classic study in 1968 proposed that bone marrow-dwelling promonocytes differentiate to monocytes, which then intravasate, circulate, and, on tissue entry, differentiate to sessile macrophages. Since then, understanding of the macrophage family relationship has undergone substantial enhancement and occasional revision. It is currently recognized that in addition to their role in the bone marrow, hematopoietic progenitors circulate and give rise to their descendants in extramedullary niches. Monocytes, of which there are several subsets, are not merely circulating macrophage precursors but participate in the immune response in their own right. Macrophages are highly heterogeneous and, as recent studies indicate, can arise in the absence of a monocyte intermediate. These spatial and developmental relationships reveal a complex interactive network and underscore the importance of context in evaluating biological systems. The observations have significant implications for how we image, target, and treat disease.
Adventitial Biology
Authors: Hu Yanhua , Xu Qingbo
Published: June 2011

Recent evidence indicates that stem/progenitor cells are present in the adventitia and participate in vascular repair and the formation of neointimal lesions in severely damaged vessels. Data have also demonstrated that these resident stem/progenitor cells could differentiate into endothelial or smooth muscle cells in response to different stimuli. Under pathological conditions, adventitial inflammation results in releasing a panel of cytokines, such as stromal cell-derived factor-1 and tumor necrosis factor-α, that may lead to local stem/progenitor mobilization and differentiation. Overall, these data support the impact of the adventitial progenitors in pathophysiological processes of lesion development in the arterial wall. In the present review, we aim to summarize the data concerning the presence of the resident stem cells and discuss the pathological impact of the adventitia in vascular diseases. We will also discuss the possible signal pathways orchestrating stem cell differentiation toward vascular lineage and highlight controversial issues related to the role of adventitial progenitors.
Loss of Methyl-CpG–Binding Domain Protein 2 Enhances Endothelial Angiogenesis and Protects Mice Against Hind-Limb Ischemic Injury
Authors: Rao Xiaoquan , Zhong Jixin , Zhang Shu , Zhang Yushan , Yu Qilin , Yang Ping , Wang Mong-Heng , Fulton David J. , Shi Huidong , Dong Zheng , Wang Daowen , Wang Cong-Yi
Published: June 2011

Background—

Despite intensive investigation, how DNA methylation influences endothelial function remains poorly understood. We used methyl-CpG–binding domain protein 2 (MBD2), an interpreter for DNA methylome–encoded information, to dissect the impact of DNA methylation on endothelial function in both physiological and pathophysiological states.

Methods and Results—

Human umbilical vein endothelial cells under normal conditions express moderate levels of MBD2, but knockdown of MBD2 by siRNA significantly enhanced angiogenesis and provided protection against H
2
O
2
-induced apoptosis. Remarkably,
Mbd2
−/−
mice were protected against hind-limb ischemia evidenced by the significant improvement in perfusion recovery, along with increased capillary and arteriole formation. Loss of MBD2 activated endothelial survival and proangiogenic signals downstream of vascular endothelial growth factor signaling characterized by an increase in endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor receptor 2 expression, along with enhanced extracellular signal-regulated kinase 1/2 activation and BCL-2 expression. Mechanistic studies confirmed the methylation of CpG elements in the eNOS and vascular endothelial growth factor receptor 2 promoter. MBD2 binds to these methylated CpG elements and suppresses eNOS promoter activity. On ischemic insult, key endothelial genes such as eNOS and vascular endothelial growth factor receptor 2 undergo a DNA methylation turnover, and MBD2 interprets the changes of DNA methylation to suppress their expressions. Moreover, MBD2 modulation of eNOS expression is likely confined to endothelial cells because nonendothelial cells such as splenocytes fail to express eNOS after loss of MBD2.

Conclusions—

We provided direct evidence supporting that DNA methylation regulates endothelial function, which forms the molecular basis for understanding how environmental insults (epigenetic factor) affect the genome to modify disease susceptibility. Because MBD2 itself does not affect the methylation of DNA and is dispensable for normal physiology in mice, it could be a viable epigenetic target for modulating endothelial function in disease states.
Mitral Valve Abnormalities Identified by Cardiovascular Magnetic Resonance Represent a Primary Phenotypic Expression of Hypertrophic Cardiomyopathy
Authors: Maron Martin S. , Olivotto Iacopo , Harrigan Caitlin , Appelbaum Evan , Gibson C. Michael , Lesser John R. , Haas Tammy S. , Udelson James E. , Manning Warren J. , Maron Barry J.
Published: June 2011

Background—

Whether morphological abnormalities of the mitral valve represent part of the hypertrophic cardiomyopathy (HCM) disease process is unresolved. Therefore, we applied cardiovascular magnetic resonance to characterize mitral valve morphology in a large HCM cohort.

Methods and Results—

Cine cardiac magnetic resonance images were obtained in 172 HCM patients (age, 42±18 years; 62% men) and 172 control subjects. In addition, 15 HCM gene-positive/phenotype-negative relatives were studied. Anterior mitral leaflet (AML) and posterior mitral leaflet lengths were greater in HCM patients than in control subjects (26±5 versus 19±5 mm,
P
<0.001; and 14±4 versus 10±3 mm,
P
<0.001, respectively), including 59 patients (34%) in whom AML length alone, posterior mitral leaflet length alone, or both were particularly substantial (>2 SDs above controls). Leaflet length was increased compared with controls in virtually all HCM age groups, including young patients 15 to 20 years of age (AML, 26±5 versus 21±4 mm;
P
=0.0002) and those ≥60 years of age (AML, 26±4 versus 19±2 mm;
P
<0.001). No relation was evident between mitral leaflet length and LV thickness or mass index (
P
=0.09 and
P
=0.16, respectively). A ratio of AML length to LV outflow tract diameter of >2.0 was associated with subaortic obstruction (
P
=0.001). In addition, AML length in 15 genotype-positive relatives without LV hypertrophy exceeded that of matched control subjects (21±3 versus 18±3 mm;
P
<0.01).

Conclusions—

In HCM, mitral valve leaflets are elongated independently of other disease variables, likely constituting a primary phenotypic expression of this heterogeneous disease, and are an important morphological abnormality responsible for LV outflow obstruction in combination with small outflow tract dimension. These findings suggest a novel role for cardiac magnetic resonance in the assessment of HCM.
Nonmuscle Myosin Light-Chain Kinase Deficiency Attenuates Atherosclerosis in Apolipoprotein E–Deficient Mice via Reduced Endothelial Barrier Dysfunction and Monocyte Migration
Authors: Sun Chongxiu , Wu Mack H. , Yuan Sarah Y.
Published: June 2011

Background—

Endothelial dysfunction and monocyte migration are key events in the pathogenesis of atherosclerosis. Nonmuscle myosin light-chain kinase (nmMLCK), the predominant MLCK isoform in endothelial cells, has been shown to contribute to vascular inflammation by altering endothelial barrier function. However, its impact on atherogenesis remains unknown.

Methods and Results—

We investigated the role of nmMLCK in the development of atherosclerotic lesions in apolipoprotein E–deficient (
apoE
−/−
) mice fed an atherogenic diet for 12 weeks. Histopathological examination demonstrated that nmMLCK deficiency (
apoE
−/−
nmmlck
−/−
) reduced the size of aortic lesions by 53%, lipid contents by 44%, and macrophage deposition by 40%. Western blotting and reverse-transcription polymerase chain reaction revealed the expression of nmMLCK in aortic endothelial cells and peripheral blood monocytes. Measurements of transendothelial electric resistance indicated that nmMLCK deficiency attenuated endothelial barrier dysfunction caused by thrombin, oxidized low-density lipoprotein, and tumor necrosis factor α. In monocytes, nmMLCK deficiency reduced their migration in response to the chemokine monocyte chemoattractant protein-1. Further mechanistic studies showed that nmMLCK acted through both myosin light chain phosphorylation-coupled and -uncoupled pathways; the latter involved Rous sacracoma virus homolog genes-encoded tyrosine kinases (Src) signaling. Moreover, depletion of Src via gene silencing, site-specific mutagenesis, or pharmacological inhibition of Src greatly attenuated nmMLCK-dependent endothelial barrier dysfunction and monocyte migration.

Conclusions—

Nonmuscle myosin light-chain kinase contributes to atherosclerosis by regulating endothelial barrier function and monocyte migration via mechanisms involving not only kinase-mediated MLC phosphorylation but also Src activation.
Extracorporeal Membrane Oxygenation for Bridge to Heart Transplantation Among Children in the United States
Authors: Almond Christopher S. , Singh Tajinder P. , Gauvreau Kimberlee , Piercey Gary E. , Fynn-Thompson Francis , Rycus Peter T. , Bartlett Robert H. , Thiagarajan Ravi R.
Published: June 2011

Background—

Extracorporeal membrane oxygenation (ECMO) has served for >2 decades as the standard of care for US children requiring mechanical support as a bridge to heart transplantation. Objective data on the safety and efficacy of ECMO for this indication are limited. We describe the outcomes of ECMO as a bridge to heart transplantation to serve as performance benchmarks for emerging miniaturized assist devices intended to replace ECMO.

Methods and Results—

Data from the Extracorporeal Life Support Organization Registry and the Organ Procurement Transplant Network database were merged to identify children supported with ECMO and listed for heart transplantation from 1994 to 2009. Independent predictors of wait-list and posttransplantation in-hospital mortality were identified. Objective performance goals for ECMO were developed. Of 773 children, the median age was 6 months (interquartile range, 1 to 44 months); 28% had cardiomyopathy; and in 38%, a bridge to transplantation was intended at ECMO initiation. Overall, 45% of subjects reached transplantation, although one third of those transplanted died before discharge; overall survival to hospital discharge was 47%. Wait-list mortality was independently associated with congenital heart disease, cardiopulmonary resuscitation before ECMO, and renal dysfunction. Posttransplantation mortality was associated with congenital heart disease, renal dysfunction, ECMO duration of >14 days, and initial ECMO indication as a bridge to recovery. In the objective performance goal cohort (n=485), patients with cardiomyopathy had the highest survival to hospital discharge (63%), followed by patients with myocarditis (59%), 2-ventricle congenital heart disease (44%) and 1-ventricle congenital heart disease (33%).

Conclusion—

Although ECMO is effective for short-term circulatory support, it is not reliable for the long-term circulatory support necessary for children awaiting heart transplantation. Fewer than half of patients bridged with ECMO survive to hospital discharge. More effective modalities for chronic circulatory support in children are urgently needed.
Relaxin Ameliorates Hypertension and Increases Nitric Oxide Metabolite Excretion in Angiotensin II But Not
N
ω
-Nitro-
l
-Arginine Methyl Ester Hypertensive Rats
Authors: Sasser Jennifer M. , Molnar Miklos , Baylis Chris
Published: June 2011

Previous findings suggest a potential therapeutic action of relaxin, the putative vasodilatory signal of normal pregnancy, in some forms of cardiovascular disease. However, the mechanisms underlying the beneficial effects of relaxin have not been fully elucidated. The purpose of this study was to determine whether the vasodilatory effects of relaxin are dependent on activation of NO synthase. We examined the effect of relaxin in male Sprague-Dawley rats given angiotensin II (Ang II; 200 ng/kg per minute SC by minipump), the NO synthase inhibitor
N
ω
-nitro-
l
-arginine methyl ester (
l
-NAME; 1.5 mg/100 g IV followed by 150 mg/L in drinking water), or vehicle for 3 weeks. After 7 days of Ang II or
l
-NAME, mean arterial pressure was elevated compared with baseline. Relaxin was administered (4 μg/h, SC by minipump) for the next 2 weeks of Ang II,
l
-NAME, or vehicle treatment. Two-week relaxin treatment alone slightly reduced mean arterial pressure in normotensive rats. Three weeks of either Ang II or
l
-NAME treatment alone produced hypertension, albuminuria, mild glomerular sclerosis, reduced nitric oxide metabolite excretion, and increased oxidative stress (excretion of hydrogen peroxide and thiobarbituric acid reactive substances and renal cortex nitrotyrosine abundance). Relaxin reduced mean arterial pressure, albumin excretion, and oxidative stress markers and preserved glomerular structure and nitric oxide metabolite excretion in Ang II–treated rats; however, relaxin did not attenuate these changes in the rats treated with
l
-NAME. None of the treatments affected protein abundance of neuronal or endothelial NO synthase in the kidney cortex. These data suggest that the vasodilatory effects of relaxin are dependent on a functional NO synthase system and increased NO bioavailability possibly because of a reduction in oxidative stress.
Ethnic Differences in Aortic Pulse Wave Velocity Occur in the Descending Aorta and May Be Related to Vitamin D
Authors: Rezai Mohammad-Reza , Wallace A. Michael , Sattar Naveed , Finn Joseph D. , Wu Frederick C.W. , Cruickshank J. Kennedy
Published: June 2011

We studied aortic pulse wave velocity (aPWV), a predictor of cardiovascular events independent of blood pressure, in a multiethnic sample of British men, to investigate the roles for blood levels of vitamin D and aldosterone in total and regional aortic stiffness. Total aPWV was estimated noninvasively by the Arteriograph device (aPWV
AG
) in 198 men, with its length measure calibrated by magnetic resonance. PWVs over the aortic arch and descending aorta were measured by magnetic resonance in a subsample (n=47). Mean (SE) aPWV
AG
in South Asians (n=68; age 55±10 years), at known higher coronary disease risk than other groups, was 0.5 m/s (0.2 m/s) higher than in African Caribbeans (n=67; 55±10 years), at lowest coronary disease risk here, and Europeans (n=63; 57±8 years), adjusted for age, systolic blood pressure, and diabetes mellitus (
P
=0.01). By magnetic resonance, PWV over the descending aorta in South Asians was 0.7 m/s (0.3 m/s) and 0.8 m/s (0.3 m/s) higher than in African Caribbeans and Europeans, respectively; PWV over the aortic arch was not different. South Asians and African Caribbeans had 21 nmol/L (3 nmol/L) and 14 nmol/L (3 nmol/L) lower mean (SE) 25(OH)D than Europeans (
P
<0.001). Unlike aldosterone, 25(OH)D was negatively correlated with aPWV
AG
adjusted for age and systolic blood pressure, as well as weakened or removed ethnic differences in aPWV
AG
in regression models. These data suggest that aortic stiffness as aPWV parallels coronary disease risk in ethnic groups, descending aortic but not arch PWV has this feature, and serum 25(OH)D is an independent negative correlate of aPWV and may partly account for ethnicity-related differences in aPWV and coronary disease risk.
Rapid and Highly Resolving: Affective Evaluation of Olfactorily Conditioned Faces
Authors: Steinberg Christian , Dobel Christian , Schupp Harald T. , Kissler Johanna , Elling Ludger , Pantev Christo , Junghöfer Markus
Published: June 2011

Evidence from hemodynamic and electrophysiological measures suggests that the processing of emotionally relevant information occurs in a spatially and temporally distributed affective network. ERP studies of emotional stimulus processing frequently report differential responses to emotional stimuli starting around 120 msec. However, the involvement of structures that seem to become activated at earlier latencies (i.e., amygdala and OFC) would allow for more rapid modulations, even in distant cortical areas. Consistent with this notion, recent ERP studies investigating associative learning have provided evidence for rapid modulations in sensory areas earlier than 120 msec, but these studies either used simple and/or very few stimuli. The present study used high-density whole-head magneto-encephalography to measure brain responses to a multitude of neutral facial stimuli, which were associated with an aversive or neutral odor. Significant emotional modulations were observed at intervals of 50–80 and 130–190 msec in frontal and occipito-temporal regions, respectively. In the absence of contingency awareness and with only two learning instances, a remarkable capacity for emotional learning is observed.