bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin‐1β induces differentiation of human mesenchymal stem cells into osteoblasts via the Wnt‐5a/receptor tyrosine kinase–like orphan receptor 2 pathway
Authors: Sonomoto Koshiro , Yamaoka Kunihiro , Oshita Koichi , Fukuyo Shunsuke , Zhang Xiangmei , Nakano Kazuhisa , Okada Yosuke , Tanaka Yoshiya
Published: September 2012

Objective

Mesenchymal stem cells (MSCs) are considered to be a novel tool for the treatment of rheumatoid arthritis (RA) because of their multipotency to differentiate into osteoblasts and chondrocytes, their immunosuppressive effects, and availability. The aim of this study was to assess the mechanisms of human MSC differentiation into osteoblasts under inflammatory conditions.

Methods

Human MSCs were cultured in commercialized osteogenic induction medium with inflammatory cytokines for up to 10 days. Osteoblast differentiation was detected by alkaline phosphatase staining and messenger RNA (mRNA) expression of multiple osteoblast markers. Mineralization was assessed by alizarin red S staining.

Results

Among the various cytokines tested, interleukin‐1β (IL‐1β) induced differentiation of human MSCs into osteoblasts, which was confirmed by alkaline phosphatase activity, expression of RUNX2 mRNA, and strong alizarin red S staining. Among various molecules of the Wnt family, Wnt‐5a and receptor tyrosine kinase–like orphan receptor 2 (Ror2), a major receptor of Wnt‐5a, were significantly induced in human MSCs by IL‐1β. Silencing of either WNT5A or ROR2 by small interfering RNA with 2 different sequences reduced alkaline phosphatase activity, RUNX2 expression, and alizarin red S staining of human MSCs induced by IL‐1β.

Conclusion

IL‐1β effectively and rapidly induced human MSC differentiation into osteoblasts and mineralization, mainly through the noncanonical Wnt‐5a/Ror2 pathway. These results suggest potential benefits of IL‐1β–treated human MSCs in the treatment of damaged bone as well as in the induction of self‐renewal and self‐repair of damaged tissue, including osseous tissue.
Thrombin induces epidermal growth factor receptor transactivation and CCL2 expression in human osteoblasts
Authors: Huang Chun‐Yin , Chen Shi‐Yann , Tsai Hsiao‐Chi , Hsu Horng‐Chaung , Tang Chih‐Hsin
Published: September 2012

Objective

Thrombin is a key factor involved in the stimulation of fibrin deposition, angiogenesis, and proinflammatory processes. Abnormalities in these processes are primary features of rheumatoid arthritis (RA). The aim of this study was to investigate the intracellular signaling pathways involved in thrombin‐induced CCL2 expression in human osteoblasts.

Methods

Thrombin‐mediated CCL2 expression was assessed by quantitative polymerase chain reaction and enzyme‐linked immunosorbent assay. The mechanisms of action of thrombin in different signaling pathways were studied using Western blotting. Knockdown of protease‐activated receptor (PAR) protein was achieved by small interfering RNA (siRNA) transfection. Chromatin immunoprecipitation assays were used to study in vivo binding of c‐Jun to the CCL2 promoter. Transient transfection was used to examine activator protein 1 (AP‐1) activity.

Results

Stimulation of human primary osteoblasts and MG‐63 cells with thrombin induced CCL2 expression. PAR‐1–specific siRNA (but not other PAR siRNA) was involved in thrombin‐mediated up‐regulation of CCL2. Thrombin‐mediated CCL2 production was attenuated by the thrombin inhibitor PPACK, the protein kinase Cδ (PKCδ) inhibitor rottlerin, the c‐Src inhibitor PP2, epidermal growth factor receptor (EGFR) inhibitor AG‐1478, MEK inhibitors PD98059 and U0126, or AP‐1 inhibitors curcumin and tanshinone IIA. Stimulation of cells with thrombin increased PKCδ, c‐Src, EGFR, MEK, and ERK activation. Treatment of osteoblasts with thrombin also increased c‐Jun phosphorylation, AP‐1 luciferase activity, and c‐Jun binding to the AP‐1 element on the CCL2 promoter.

Conclusion

Our results suggest that the interaction between thrombin and PAR‐1 increases CCL2 expression in human osteoblasts via the PKCδ/c‐Src/ EGFR transactivation/MEK/ERK/c‐Jun/AP‐1 pathway.
Expression of HLA–B27 causes loss of migratory dendritic cells in a rat model of spondylarthritis
Authors: Utriainen Lotta , Firmin Dawn , Wright Pamela , Cerovic Vuk , Breban Maxime , McInnes Iain , Milling Simon
Published: September 2012

Objective

In rats transgenic for human HLA–B27 and β2‐microglobulin (B27‐transgenic rats), colitis and peripheral inflammation develop spontaneously. Therefore, B27‐transgenic rats provide a model of spondylarthritis. Because inflammation in these rats requires CD4+ T lymphocytes and involves intestinal pathology, we hypothesized that dendritic cells (DCs) that migrate from the intestine and control CD4+ T cell differentiation would be aberrant in B27‐transgenic rats.

Methods

Migrating intestinal lymph DCs were collected via thoracic duct cannulation from B27‐transgenic and control (HLA–B7–transgenic or nontransgenic) rats. The phenotypes of these DCs and of mesenteric lymph node DCs were assessed by flow cytometry. The ability of DCs to differentiate from bone marrow precursors in vitro was also assessed.

Results

Lymph DCs showed increased activation and, strikingly, lacked the specific DC population that is important for maintaining tolerance to self‐antigens. This population of DCs was also depleted from the mesenteric lymph nodes of B27‐transgenic rats. Furthermore, in vitro culture of DCs from bone marrow precursors revealed a defect in the ability of B27‐transgenic rats to produce DCs of the migratory phenotype, although the DCs that were generated induced enhanced interleukin‐17 (IL‐17) production from naive CD4+ T cells.

Conclusion

We describe 2 different mechanisms by which HLA–B27 may contribute to inflammatory disease: increased apoptotic death of B27‐transgenic DCs that normally function to maintain immunologic tolerance and enhanced IL‐17 production from CD4+ T cells stimulated by the surviving B27‐transgenic DCs.
Chondrocyte‐intrinsic Smad3 represses Runx2‐inducible matrix metalloproteinase 13 expression to maintain articular cartilage and prevent osteoarthritis
Authors: Chen Carol G. , Thuillier Daniel , Chin Emily N. , Alliston Tamara
Published: September 2012

Objective

To identify mechanisms by which Smad3 maintains articular cartilage and prevents osteoarthritis.

Methods

A combination of in vivo and in vitro approaches was used to test the hypothesis that Smad3 represses Runx2‐inducible gene expression to prevent articular cartilage degeneration. Col2‐Cre;Smad3fl/fl mice allowed study of the chondrocyte‐intrinsic role of Smad3 independently of its role in the perichondrium or other tissues. Primary articular cartilage chondrocytes from Smad3fl/fl mice and ATDC5 chondroprogenitor cells were used to evaluate Smad3 and Runx2 regulation of matrix metalloproteinase 13 (MMP‐13) messenger RNA (mRNA) and protein expression.

Results

Chondrocyte‐specific reduction of Smad3 caused progressive articular cartilage degeneration due to imbalanced cartilage matrix synthesis and degradation. In addition to reduced type II collagen mRNA expression, articular cartilage from Col2‐Cre;Smad3fl/fl mice was severely deficient in type II collagen and aggrecan protein due to excessive MMP‐13–mediated proteolysis of these key cartilage matrix constituents. Normally, transforming growth factor β (TGFβ) signals through Smad3 to confer a rapid and dynamic repression of Runx2‐inducible MMP‐13 expression. However, we found that in the absence of Smad3, TGFβ signals through p38 and Runx2 to induce MMP‐13 expression.

Conclusion

Our findings elucidate a mechanism by which Smad3 mutations in humans and mice cause cartilage degeneration and osteoarthritis. Specifically, Smad3 maintains the balance between cartilage matrix synthesis and degradation by inducing type II collagen expression and repressing Runx2‐inducible MMP‐13 expression. Selective activation of TGFβ signaling through Smad3, rather than p38, may help to restore the balance between matrix synthesis and proteolysis that is lost in osteoarthritis.
Brief Report: To squeeze or not to squeeze, that is the question! Optimizing the Disease Activity Score in 28 joints by adding the squeeze test of metatarsophalangeal joints in early rheumatoid arthritis
Authors: de Jong P. H. , Weel A. E. , de Man Y. A. , Huisman A. M. , Gerards A. H. , van Krugten M. V. , Luime J. J. , Hazes J. M.
Published: September 2012

Objective

To optimize use of the Disease Activity Score in 28 joints (DAS28) in early rheumatoid arthritis (RA) by adding the “squeeze test” of forefeet.

Methods

The squeeze test is used to examine bilateral compression pain (BCP) across the metatarsophalangeal (MTP) joints. For this study, data for patients participating in the Treatment in the Rotterdam Early Arthritis Cohort study, an ongoing clinical trial that evaluates different induction therapies in patients with early RA, were randomly divided into 2 subsets. In subset 1 (149 patients and 819 disease activity assessments), the mathematical function of the DAS28‐squeeze was constructed using a linear regression model with the DAS as the dependent variable and the DAS28 and squeeze test as the independent variables. A DAS28‐BCP disease state was also constructed, in which DAS28 disease state categorizations were upgraded one state if the result of the squeeze test was positive. In subset 2 (153 patients and 754 assessments), concordance in disease states between the DAS28, DAS28‐squeeze, and DAS28‐BCP disease states was compared, using both the DAS and Boolean‐defined remission criteria as reference.

Results

Agreement between the DAS and the DAS28‐squeeze (82%) was significantly higher than agreement between the DAS and the DAS28 (76%). When we assessed the group of patients who had arthritis of the forefeet only (22 patients and 46 assessments), overall agreement between the DAS and the DAS28 was 40%, while agreement between the DAS and the DAS28‐squeeze was 59% and that between the DAS and the DAS28‐BCP disease state was 65%. Furthermore, the specificities of the DAS28‐squeeze and the DAS28‐BCP (80% and 81%, respectively) were higher than that of the DAS28 (76%), while the sensitivities of the DAS28, DAS28‐squeeze, and DAS28‐BCP to identify true remission according to the Boolean criteria were 88%, 87%, and 81%, respectively.

Conclusion

Adding the squeeze test of forefeet to the DAS28 has value for dependably classifying the disease state in patients with early RA.
It takes time: A remarkable example of delayed recognition
Authors: Van Calster Ben
Published: September 2012

The way in which scientific publications are picked up by the research community can vary. Some articles become instantly cited, whereas others go unnoticed for some time before they are discovered or rediscovered. Papers with delayed recognition have also been labeled “sleeping beauties.” I briefly discuss an extreme case of a sleeping beauty. Peirce's short note in Science in 1884 shows a remarkable increase in citations since around 2000. The note received less than 1 citation per year in the decades prior to 2000, 3.5 citations per year in the 2000s, and 10.4 in the 2010s. This increase was seen in several domains, most notably meteorology, medical prediction research, and economics. The paper outlines formulas to evaluate a binary prediction system for a binary outcome. This citation increase in various domains may be attributed to a widespread, growing research focus on mathematical prediction systems and the evaluation thereof. Several recently suggested evaluation measures essentially reinvented or extended Peirce's 120‐year‐old ideas.
Platinum‐Coated Porous Gold Nanorods in Methanol Electrooxidation: Dependence of Catalytic Activity on Ligament Size
Authors: Yoo Sang‐Hoon , Liu Lichun , Cho Sang Hyun , Park Sungho
Published: September 2012

Here we demonstrate that, in the dealloying process of Au–Ag nanorods, temperature is the key parameter for producing porous Au nanorods with tunable ligament sizes. The vertically aligned Au–Ag alloy nanorods were first synthesized by the electrochemical co‐deposition of Au and Ag onto anodic aluminum oxide (AAO) membrane templates. Porous Au nanorods were then obtained by selectively etching Ag away from the precursor Au–Ag alloy nanorods. Control of the ligament size was achieved by controlling the dealloying temperature. Pt deposited on the porous Au nanorods with smaller ligaments exhibited a higher catalytic activity during methanol electrooxidation than those deposited on nanorods with larger ligaments produced by dealloying at higher temperatures. The strong dependence of the catalytic activity on the ligament size of porous Au is principally due to different amounts of carbon monoxide (CO) generated during methanol electrooxidation. Less CO was generated as the ligament size decreased. This finding is of importance for developing highly efficient cathode materials for carrying out methanol electrooxidation in practical applications in which porous Au with a large surface area is used as a supporting substrate.
A consideration of the evidence that genetic defects in planar cell polarity contribute to the etiology of human neural tube defects
Authors: Juriloff Diana M. , Harris Muriel J.
Published: September 2012

A variety of human birth defects originate in failure of closure of the embryonic neural tube. The genetic cause of the most common nonsyndromic defects, spina bifida (SB) or anencephaly, is considered to be combinations of variants at multiple genes. The genes contributing to the etiology of neural tube closure defects (NTDs) are unknown. Mutations in planar cell polarity (PCP) genes in mice cause a variety of defects including the NTD, craniorachischisis, and sometimes SB or exencephaly (EX); they also demonstrate the role of digenic combinations of PCP mutants in NTDs. Recent studies have sought rare predicted‐to‐be‐deleterious alterations (putative mutations) in coding sequence of PCP genes in human cases with various anomalies of the neural tube. This review summarizes the cumulative results of these studies according to a framework based on the embryopathogenesis of NTDs, and considers some of the insights from the approaches used and the limitations. Rare putative mutations in the PCP genesVANGL2,SCRIB,DACT1, andCELSR1cumulatively contributed to over 20% of cases with craniorachischisis, a rare defect; no contributing variants were found forPRICKLE1orPTK7. PCP rare putative mutations had a weaker role in myelomeningocele (SB), being found in approximately 6% of cases and cumulated acrossCELSR1,FUZ,FZD6,PRICKLE1,VANGL1, andVANGL2. These results demonstrate that PCP gene alterations contribute to the etiology of human NTDs. We recommend that future research should explore other types of PCP gene variant such as regulatory mutations and low frequency (1 to 5%) deleterious polymorphisms. Birth Defects Research (Part A) 2012. © 2012 Wiley Periodicals, Inc.
Transcriptional analyses of two mouse models of spina bifida
Authors: Cabrera Robert M. , Finnell Richard H. , Zhu Huiping , Shaw Gary M. , Wlodarczyk Bogdan J.
Published: September 2012

BACKGROUND

Spina bifida is one of the most common of all human structural birth defects. Despite considerable effort over several decades, the causes and mechanisms underlying this malformation remain poorly characterized.

METHODS

To better understand the pathogenesis of this abnormality, we conducted a microarray study using Mouse Whole Genome Bioarrays which have ∼36,000 gene targets, to compare gene expression profiles between two mouse models; CXL‐Splotch and FKBP8Gt(neo) which express a similar spina bifida phenotype. We anticipated that there would be a collection of overlapping genes or shared genetic pathways at the molecular level indicative of a common mechanism underlying the pathogenesis of spina bifida during embryonic development.

RESULTS

A total of 54 genes were determined to be differentially expressed (25 downregulated, 29 upregulated) in the FKBP8Gt(neo) mouse embryos; whereas 73 genes were differentially expressed (56 downregulated, 17 upregulated) in the CXL‐Splotch mouse relative to their wild‐type controls. Remarkably, the only two genes that showed decreased expression in both mutants were v‐ski sarcoma viral oncogene homolog (Ski), and Zic1, a transcription factor member of the zinc finger family. Confirmation analysis using quantitative real‐time (qRT)‐PCR indicated that only Zic1 was significantly decreased in both mutants. Gene ontology analysis revealed striking enrichment of genes associated with mesoderm and central nervous system development in the CXL‐Splotch mutant embryos, whereas in the FKBP8Gt(neo) mutants, the genes involved in dorsal/ventral pattern formation, cell fate specification, and positive regulation of cell differentiation were most likely to be enriched. These results indicate that there are multiple pathways and gene networks perturbed in mouse embryos with shared phenotypes. Birth Defects Research (Part A) 2012. © 2012 Wiley Periodicals, Inc.
Systematic review of outcomes of liver resection for early hepatocellular carcinoma within the Milan criteria
Authors: Lim K-C , Chow P K-H , Allen J C , Siddiqui F J , Chan E S-Y , Tan S-B
Published: September 2012

Background

Long-term overall survival after liver resection in patients with hepatocellular carcinoma (HCC) within the Milan criteria has been reported to improve in recent years. This study systematically reviewed the outcomes of surgical resection for HCC in patients with good liver function and meeting the Milan criteria for early HCC, published in the past 10 years.

Methods

A literature search was conducted in PubMed for papers on outcomes of surgical resection for HCC published between January 2000 and December 2010. Cochrane systematic review methodology was used for this review. The primary outcome was overall survival. Secondary outcomes included operative mortality and disease-free survival. Studies that focused on geriatric populations, paediatric populations, a subset of the Milan criteria (such solitary tumours) or included patients with incidental tumours were excluded, as were case reports, conference abstracts, and studies with a large proportion of Child–Pugh grade C liver cirrhosis or unknown Child–Pugh status.

Results

Of 152 studies reviewed, two randomized clinical trials and 27 retrospective case series were eligible for inclusion. The 5-year overall survival rate after resection of HCC ranged from 27 to 81 (median 67) per cent, and the median disease-free survival rate from 21 to 57 (median 37) per cent. There was a trend towards improved overall survival in recent years. The operative mortality rate ranged from 0 to 5 (median 0·7) per cent.

Conclusion

Surgical resection offers good overall survival for patients with HCC within the Milan criteria and with good liver function, although recurrence rates remain high. Outcomes have tended to improve in more recent years.
Predictive factors for lymph node metastasis in patients with poorly differentiated early gastric cancer
Authors: Lee J H , Choi M-G , Min B-H , Noh J H , Sohn T S , Bae J M , Kim S
Published: September 2012

Background

Endoscopic submucosal dissection is gaining popularity in the treatment of early gastric cancer. This study aimed to identify clinicopathological factors predictive of lymph node metastasis in patients with the poorly differentiated early gastric cancer to assess the feasibility of using endoscopic submucosal dissection for these cancers.

Methods

The records of patients with poorly differentiated early gastric cancer who had undergone gastric cancer surgery between January 2002 and December 2009 were reviewed. Associations between clinicopathological factors and the presence of lymph node metastasis were analysed by univariable and multivariable logistic regression analysis.

Results

Some 1005 patients were included in the analysis. Univariable analysis indicated that lymph node metastasis was associated with sex, ulceration, tumour size, depth of invasion, macroscopic type, lymphatic invasion and venous invasion. Logistic regression revealed that lymph node metastasis was significantly associated with sex, tumour size, depth of tumour invasion and lymphatic involvement. In the group with none of these risk factors (men with mucosal tumour no larger than 2 cm in size, with no lymphatic involvement), lymph node metastasis was present in four (3·2 per cent) of 124 patients.

Conclusion

In the present study 3·2 per cent of patients who were negative for all identified risk factors had lymph node metastasis. The use of endoscopic submucosal dissection should be considered carefully in the treatment of poorly differentiated early gastric cancer.
Facilitating surgical research across Europe
Authors: Scholefield J H , Lobo D N
Published: September 2012

Frameworks are in place to facilitate surgical trials
Quantitative determination of paclitaxel and its metabolites, 6α‐hydroxypaclitaxel and p‐3′‐hydroxypaclitaxel, in human plasma using column‐switching liquid chromatography/tandem mass spectrometry
Authors: Yamaguchi Hiroaki , Fujikawa Asuka , Ito Hajime , Tanaka Nobuaki , Furugen Ayako , Miyamori Kazuaki , Takahashi Natsuko , Ogura Jiro , Kobayashi Masaki , Yamada Takehiro , Mano Nariyasu , Iseki Ken
Published: September 2012

A column‐switching liquid chromatography/electrospray ionization tandem mass spectrometry to determine paclitaxel and its metabolites, 6α‐hydroxypaclitaxel and p‐3′‐hydroxypaclitaxel, in human plasma was developed. The analytical system had a Shim‐Pack MAYI‐ODS (10 × 4.6 mm i.d.) trapping column with deproteinization ability that concentrates analytes and removes water‐soluble components. This method covered a linearity range of 5–5000 ng/mL of concentrations in plasma for paclitaxel, a range of 0.87–870 ng/mL for 6α‐hydroxypaclitaxel and a range of 0.87–435 ng/mL for p‐3′‐hydroxypaclitaxel. The intra‐day precision and inter‐day precision of analysis were less than 11.1%, and the accuracy was within ±14.4% at concentrations of 5, 50, 500 and 5000 ng/mL for paclitaxel, 0.87, 8.7, 87 and 870 ng/mL for 6α‐hydroxypaclitaxel, and 0.87, 8.7, 87 and 435 ng/mL for p‐3′‐hydroxypaclitaxel. The total run time was 30 min. Our method was successfully applied to clinical pharmacokinetic investigation. Copyright © 2012 John Wiley & Sons, Ltd.
Determination of cnidilin and its two metabolites in rat bile and stool after oral administration by HPLC/electrospray ionization tandem mass spectrometry
Authors: Zhu Hong , Ren Yanping , Sun Yingguang , Chang Lu , Cao Liang , Xu Huijin , Zhang Lantong
Published: September 2012

A simple, fast and sensitive method for the simultaneous determination of cnidilin and its two metabolites (M1 and M2) in rat bile and stool using HPLC coupled with electrospray ionization tandem mass spectrometry (HPLC‐ESI‐MS/MS) has been developed. The sample pretreatment was simple, because methanol was the only additive used for dilution of bile and ultrasound of stool. Pimpinellin was used as internal standard (IS). The separation was performed on a reverse phase C18 column with gradient elution consisting of 0.5‰ aqueous formic acid and methanol (containing 0.5‰ formic acid). The detection was in the multiple‐reaction monitoring mode within 7 min. All the analytes were in accordance with the requirement of the validation of the method in vivo (linearity, precision, accuracy, limit of detection and limit of quantification). After oral administrating 24 mg/kg of the prototype drug cnidilin, M1 and M2 were determined in bile within 36 h, and in stool within 60 h. Cnidilin in bile was completely excreted in 24 h, and the main excretive amount of cnidilin was 80% in the first 6 h, but the drug recovery in bile within 24 h was <1.95%. In stool, the main excretive amount of cnidilin was 95.8% in the first 24 h, and the drug recovery within 48 h was lower than 1.48%. Copyright © 2012 John Wiley & Sons, Ltd.
Evolutionary Imprint of Catalytic Domains in Fungal PKS–NRPS Hybrids
Authors: Boettger Daniela , Bergmann Holger , Kuehn Barbara , Shelest Ekaterina , Hertweck Christian
Published: September 2012

Fungal hybrid enzymes consisting of a polyketide synthase (PKS) and a nonribosomal peptide synthetase (NRPS) module are involved in the biosynthesis of a vast array of ecologically and medicinally relevant natural products. Whereas a dozen gene clusters could be assigned to the requisite PKS–NRPS pathways, the programming of the multifunctional enzymes is still enigmatic. Through engineering and heterologously expressing a chimera of PKS (lovastatin synthase, LovB) and NRPS (cytochalasin synthase, CheA) in Aspergillus terreus, we noted the potential incompatibility of a fungal highly reducing PKS (hrPKS) with the NRPS component of fungal PKS–NRPS hybrids. To rationalize the unexpected outcome of the gene fusion experiments, we conducted extensive bioinformatic analyses of fungal PKS–NRPS hybrids and LovB‐type PKS. From motif studies and the function of the engineered chimeras, a noncanonical function of C‐terminal condensation (C) domains in truncated PKS–NRPS homologues was inferred. More importantly, sequence alignments and phylogenetic trees revealed an evolutionary imprint of the PKS–NRPS domains, which reflect the evolutionary history of the entire megasynthase. Furthermore, a detailed investigation of C and adenylation (A) domains provides support for a scenario in which not only the A domain but also the C domain participates in amino acid selection. These findings shed new light on the complex code of this emerging class of multifunctional enzymes and will greatly facilitate future combinatorial biosynthesis and pathway engineering approaches towards natural product analogues.
Unusual CC Bond Isomerization of an α,β‐Unsaturated γ‐Butyrolactone Catalysed by Flavoproteins from the Old Yellow Enzyme Family
Authors: Durchschein Katharina , Wallner Silvia , Macheroux Peter , Zangger Klaus , Fabian Walter M. F. , Faber Kurt
Published: September 2012

An unexpected, redox‐neutral CC bond isomerization of a γ‐butyrolactone bearing an exo‐methylene unit to the thermodynamically more favoured endo isomer (kcat=0.076 s−1) catalysed by flavoproteins from the Old Yellow Enzyme family was discovered. Theoretical calculations and kinetic data support a mechanism through which the isomerization proceeds through FMN‐mediated hydride addition onto exo‐Cβ, followed by hydride abstraction from endo‐Cβ′, which is in line with the well‐established CC bond bioreduction of OYEs. This new isomerase activity enriches the catalytic versatility of ene‐reductases.
Immediate results of bifurcational stenting assessed with optical coherence tomography
Authors: Viceconte Nicola , Tyczynski Pawel , Ferrante Giuseppe , Foin Nicolas , Chan Pak Hei , Barrero Eduardo Alegrìa , Di Mario Carlo
Published: September 2012

Background: A tubular stent may adapt with difficulty to coronary bifurcation lesions (CBLs). Methods: Time domain or frequency domain (FD) optical coherence tomography (OCT) was performed to assess strut apposition immediately after stent implantation across four segments inside the bifurcation, in a consecutive series of patients. OCT pullbacks were performed in the main vessel (MV). Results: A total of 13,142 struts in 45 CBL in 41 patients were assessed. Strut malapposition was significantly more frequent in the half bifurcation facing the side‐branch (SB) ostium (42.9%) than in the proximal segment of the bifurcation 11.8%, half bifurcation opposite the SB 6.7%, or the distal segment 5.7% (all P < 0.0001). Lesions (n = 15) treated with stenting of both MV and SB had a total higher rate of malapposition than those (n = 30) treated with stenting of the MV only (17.6% vs. 9.5%; P = 0.0014). In latter group, lesions treated with FD‐OCT‐guided stent implantation (n = 13) presented a lower rate of malapposition than those treated with conventional angiographic‐guided stent implantation (n = 17) (7.1% vs. 17.5%; P = 0.005). Conclusions: In CBL, strut malapposition is particularly high at the SB ostium. However, a strategy of stenting MV only with adjunctive FD‐OCT guidance is associated with lower rates of malapposition. © 2012 Wiley Periodicals, Inc.
Transcatheter “thrombin‐blood patch” injection: A novel and effective approach to treat catheterization‐related arterial perforation
Authors: Maluenda Gabriel , Mitulescu Lavinia , Ben‐Dor Itsik , Sardi Gabriel , Romaguera Rafael , Satler Lowell F. , Pichard Augusto D. , Waksman Ron , Bernardo Nelson L.
Published: September 2012

Objective

This study aimed to describe the safety and feasibility of transcatheter “thrombin‐blood patch” (TBP) injection to treat catheterization‐related arterial vascular access perforation.

Background

Vascular access complications are infrequent but potentially life threatening conditions related to percutaneous procedures. Surgical vascular repair are associated with high rates of morbidity and mortality due to advanced cardiovascular disease.

Methods

From October 2007 to July 2010 we studied 23 patients who presented active access arterial bleeding after percutaneous procedures and underwent transcatheter angiographic guided TBP injection across the entry site of the arterial perforation as a primary approach.

Results

The mean age of the population was 67 years, predominantly female (78.3%) with high rate of comorbidities including diabetes (30.4%), prior coronary revascularization (50.0%), chronic renal failure (43.5%), and heart failure (56.5%). Thirteen patients (56.5%) developed severe hypotension after the index procedure. The repair procedure had a mean duration of 82 ± 57 minutes. TBP was injected in all patients. One case additionally required covered‐stent to obtain hemostasis. Angiographic success was achieved in the 23 patients; however, one case required a second intervention due to recurrent bleeding, which was effectively treated using covered‐stent. All patients were discharged alive and no major cardiovascular events, including myocardial infarction/stroke, were observed.

Conclusions

Transcatheter “thrombin‐blood patch” injection is a safe, novel technique that allows prompt percutaneous approach to treat catheterization‐related arterial perforation. This strategy appears particularly attractive to treat patients who cannot tolerate “open” vascular reconstruction and repair. © 2012 Wiley Periodicals, Inc.
Smoking status and survival in the national comprehensive cancer network non–small cell lung cancer cohort
Authors: Ferketich Amy K. , Niland Joyce C. , Mamet Rizvan , Zornosa Carrie , D'Amico Thomas A. , Ettinger David S. , Kalemkerian Gregory P. , Pisters Katherine M. , Reid Mary E. , Otterson Gregory A.
Published: September 2012

BACKGROUND:

The objectives of this study were to evaluate survival among current smokers, former smokers, and never smokers who are diagnosed with non–small cell lung cancer (NSCLC).

METHODS:

The study included patients who participated in the National Comprehensive Cancer Network's NSCLC Database Project. Current, former, and never smokers were compared with respect to overall survival by fitting Cox regression models.

RESULTS:

Data from 4200 patients were examined, including 618 never smokers, 1483 current smokers, 380 former smokers who quit 1 to 12 months before diagnosis, and 1719 former smokers who quit >12 months before diagnosis. Among patients with stage I, II, and III disease, only never smokers had better survival than current smokers (hazard ratio, 0.47 [95% confidence interval, 0.26‐0.85] vs 0.51 [95% confidence interval, 0.38‐0.68], respectively). Among patients with stage IV disease, the impact of smoking depended on age: Among younger patients (aged ≤55 years), being a never smoker and a former smoker for ≥12 months increased survival. After age 85 years, smoking status did not have a significant impact on overall survival.

CONCLUSIONS:

Patients who were smoking at the time of diagnosis had worse survival compared with never smokers. Among younger patients with stage IV disease, current smokers also had worse survival compared with former smokers who quit >12 months before diagnosis. It is likely that tumor biology plays a major role in the differences observed; however, to improve survival, it is prudent to encourage all smokers to quit smoking if they are diagnosed with NSCLC. Cancer 2013. © 2012 American Cancer Society.
Diffusion‐Barrier‐Free Porous Carbon Monoliths as a New Form of Activated Carbon
Authors: Kubo Takashi , Sakamoto Hirotoshi , Fujimori Toshihiko , Itoh Tsutomu , Ohba Tomonori , Kanoh Hirofumi , Martínez‐Escandell Manuel , Ramos‐Fernández José M. , Casco Mirian , Rodríguez‐Reinoso Francisco , Urita Koki , Moriguchi Isamu , Endo Morinobu , Kaneko Katsumi
Published: September 2012

For the practical use of activated carbon (AC) as an adsorbent of CH4, tightly packed monoliths with high microporosity are supposed to be one of the best morphologies in terms of storage capacity per apparent volume of the adsorbent material. However, monolith‐type ACs may cause diffusion obstacles in adsorption processes owing to their necked pore structures among the densely packed particles, which result in a lower adsorption performance than that of the corresponding powder ACs. To clarify the relationship between the pore structure and CH4 adsorptivity, microscopic observations, structural studies on the nanoscale, and conductivity measurements (thermal and electrical) were performed on recently developed binder‐free, self‐sinterable ACs in both powder and monolithic forms. The monolith samples exhibited higher surface areas and electrical conductivities than the corresponding powder samples. Supercritical CH4 adsorption isotherms were measured for each powder and monolith sample at up to 7 MPa at 263, 273, and 303 K to elucidate their isosteric heats of adsorption and adsorption rate constants, which revealed that the morphologies of the monolith samples did not cause serious drawbacks for the adsorption and desorption processes. This will further facilitate the availability of diffusion‐barrier‐free microporous carbon monoliths as practical CH4 storage adsorbents.
A heat‐inducible CRE/LOXP gene induction system in medaka
Authors: Kobayashi Kayo , Kamei Yasuhiro , Kinoshita Masato , Czerny Thomas , Tanaka Minoru
Published: September 2012

We established three lines of transgenic medaka, a heat‐shock element (HSE) monitor line (hse‐GFP line), heat‐inducible driver lines (hse‐cre lines), and effector lines (gapdh‐loxP[DsRed]‐GFP lines). We employed these to comprehensively analyze gene induction at different time points in various tissues. These analyses demonstrate a good response of synthetic HSEs by heat treatment during embryogenesis and the mosaic gene induction by cre/loxP‐mediated recombination, thus providing practical information regarding the feasibility of a heat‐inducible cre/loxP‐mediated system in medaka. We also activated recombination by local heat‐treatment using a metal probe and an infrared laser. Our results collectively indicate that these lines allow us to perform lineage tracing and mosaic analysis and provide the platform to investigate gene functions at later developmental stage and adult. genesis 51:59–67, 2013. © 2012 Wiley Periodicals, Inc.
Effect of latitudinal gradient and impact of logging on genetic diversity of Cedrela lilloi along the Argentine Yungas Rainforest
Authors: Inza Maria V. , Zelener Noga , Fornes Luis , Gallo Leonardo A.
Published: September 2012

Cedrela lilloi C. DC. (cedro coya, Meliaceae), an important south American timber species, has been historically overexploited through selective logging in Argentine Yungas Rainforest. Management and conservation programs of the species require knowledge of its genetic variation patterns; however, no information is available. Molecular genetic variability of the species was characterized to identify high‐priority populations for conservation and domestication purposes. Fourteen native populations (160 individuals) along a latitudinal gradient and with different logging's intensities were assessed by 293 polymorphic AFLP (amplified fragment length polymorphism) markers. Genetic diversity was low (Ht = 0.135), according to marginal location of the species in Argentina. Most of the diversity was distributed within populations (87%). Northern populations showed significant higher genetic diversity (R2= 0.69) that agreed with latitudinal pattern of distribution of taxonomic diversity in the Yungas. Three clusters were identified by Bayesian analysis in correspondence with northern, central, and southern Yungas. An analysis of molecular variance (AMOVA) revealed significant genetic differences among latitudinal clusters even when logging (ΦRT = 0.07) and unlogging populations (ΦPT = 0.10) were separately analyzed. Loss of genetic diversity with increasing logging intensity was observed between neighboring populations with different disturbance (ΦPT = 0.03–0.10). Bottlenecks in disturbed populations are suggested as the main cause. Our results emphasize both: the necessity of maintaining the genetic diversity in protected areas that appear as possible long‐term refuges of the species; and to rescue for the national system of protected areas some high genetic diversity populations that are on private fields.
Synthesis and Photoisomerization of Rhodopsin‐Based Molecular Switches
Authors: Blanco‐Lomas Marina , Campos Pedro J. , Sampedro Diego
Published: September 2012

A new family of switches based on a rhodopsin chromophore is presented. These new photoswitches fulfil most of the required features for an efficient switch to be used in practical applications. The synthetic route is simple and versatile and enables substituents capable of acting as anchoring points to be incorporated. Also, substitution can be used to fine tune the switches' properties. The photostationary state can be reached easily by using low‐energy light compatible with complex environments. Thermal reset allows these switches to operate through on/off cycles.
Metal‐organic framework ZIF‐8 nanocrystals as pseudostationary phase for capillary electrokinetic chromatography
Authors: Li Li‐Man , Wang He‐Fang , Yan Xiu‐Ping
Published: September 2012

The outstanding properties such as large surface area, diverse structure, and accessible tunnels and cages make metal organic frameworks (MOFs) attractive as novel separation media in separation sciences. However, the utilization of MOFs in EKC has not been reported before. Here we show the exploration of zeolitic imidazolate framework‐8 (ZIF‐8), one of famous MOFs, as the pseudostationary phase (PSP) in EKC. ZIF‐8 nanocrystals were used as the PSP through dispersing in the running buffer (20 mM phosphate solution containing a 1% v/v methanol (pH 9.2)) to enhance the separation of the phenolic isomers (p‐benzenediol, m‐benzenediol, o‐benzenediol, m‐nitrophenol, p‐nitrophenol, and o‐nitrophenol). ZIF‐8 nanocrystals in the running buffer were negatively charged, and interacted with the phenolic hydroxyl groups of the analytes, and thus greatly improved the separation of the phenolic isomers. Inclusion of 200 mg L−1 ZIF‐8 in the running buffer as the background electrolyte gave a baseline separation of the phenolic isomers within 4 min. The relative standard deviations for five replicate separations of the phenolic isomers were 0.2–1.1% for migration time and 4.5–9.7% for peak area. The limits of detection varied from 0.44 to 2.0 mg L−1. The results show that nanosized MOFs are promising for application in EKC.
Progress toward the application of molecular force spectroscopy to DNA sequencing
Authors: Cheng Peng , Oliver Piercen M. , Barrett Michael J. , Vezenov Dmitri
Published: September 2012

Many recent advances in DNA sequencing have taken advantage of single‐molecule techniques using fluorescently labeled oligonucleotides as the principal mode of detection. However, in spite of the successes of fluorescent‐based sequencers, avoidance of labeled nucleotides could substantially reduce the costs of sequencing. This article discusses the development of an alternative sequencing method in which unlabeled DNA can be manipulated directly on a massively parallel scale using single‐molecule force spectroscopy. We combine a wide‐field optical detection technique (evanescent field excitation) with one of two methods of applying force in parallel, magnetic or dielectrophoretic tweezers, to attain near single‐base sensitivity in the double‐stranded character of DNA. This article will discuss the developments of such a single‐molecule force spectroscopy technique as a potential technology for genome sequencing.
Triblock copolymer‐based microchip device for rapid analysis of stuffer‐free multiplex ligation‐dependent probe amplification products
Authors: Shin Gi Won , Kim Yong Tae , Heo Hyun Young , Chung Boram , Seo Tae Seok , Jung Gyoo Yeol
Published: September 2012

Recent improvements in the multiplex ligation‐dependent probe amplification (MLPA) method promise successful multiplex analysis of various genetic markers. In particular, it has been demonstrated that elimination of the stuffer sequence included in MLPA probes for length‐dependent analysis substantially simplifies the probe design process and improves the accuracy of the analysis. As is the case for other CE‐based methods, MLPA could be further developed on a microchip platform. However, high‐resolution analysis of short MLPA probes requires careful microchip operation. In this study, we developed a microchip device for the multiplex analysis of five food‐borne pathogens using a stuffer‐free probe set. Microchip channel design and electrophoresis operating conditions were first optimized for reproducible analysis, after which two sieving matrices were tested. Finally, the method was validated using DNA samples isolated from intentionally infected milk.
A poly (4‐vinylpridine‐co‐ethylene glycol dimethacrylate) monolithic concentrator for in‐line concentration‐capillary electrophoresis analysis of phenols in water samples
Authors: Wu Yue , Zhang Wenpeng , Chen Zilin
Published: September 2012

A poly(4‐vinylpridine‐co‐ethylene glycol dimethacrylate) monolith was synthesized in a capillary and constructed as a concentrator for the in‐line polymeric monolith microextraction coupling with capillary electrophoresis. The integrated system was then used for the simultaneous determination of five trace phenols (2‐nitrophenol, 3‐nitrophenol, 4‐nitrophenol, 2‐chlorophenol, and 2,4‐dichlorophenol) in water samples. The experimental parameters for in‐line solid‐phase extraction, such as composition and volume of the elution plug, pH of sample solution, and the time for sample loading were optimized. The sensitivity for the mixture of phenols (2‐nitrophenol, 3‐nitrophenol, 4‐nitrophenol, 2‐chlorophenol, and 2,4‐dichlorophenol) enhanced to 615–2222 folds at the optimum condition was compared to the sensitivity for a normal hydrodynamic injection in capillary electrophoresis. Linearity ranged from concentration of 10–500 ng mL−1(R2 > 0.999) for all five phenols with the detection limits of 1.3–3.3 ng mL−1. In tap, snow and Yangtze River water spiked with 20 ng mL−1 and 200 ng mL−1, respectively, the recoveries of 84–105% were obtained. It has been demonstrated that this work has great potential for the analysis of phenols in genuine water samples.
Different characteristics identified by single nucleotide polymorphism array analysis in leukemia suggest the need for different application strategies depending on disease category
Authors: Huh Jungwon , Jung Chul Won , Kim Hyeoung‐Joon , Kim Yeo‐Kyeoung , Moon Joon Ho , Sohn Sang Kyun , Kim Hee‐Je , Min Woo Sung , Kim Dong Hwan (Dennis)
Published: September 2012

The purpose of this study was to evaluate the detection rate of chromosomal rearrangements in leukemia using single nucleotide polymorphism array (SNP‐A) in combination with metaphase cytogenetics (MC), with the aim of proposing a practical approach for clinical karyotyping applications of SNP‐A. The Genome‐Wide Human SNP Array 6.0 (Affymetrix, Santa Clara, CA) was applied in 469 patients with a variety of hematologic malignancies. Combined use of SNP‐A with MC improved the detection rate in comparison with MC alone: acute myeloid leukemia (AML) with normal karyotype (NK), 32% versus 0%; core binding factor (CBF)‐AML 40% versus 29%; myelodysplastic syndrome (MDS), 54% versus 39%; chronic myeloid leukemia (CML), 24% versus 3%; and acute lymphoblastic leukemia (ALL), 88% versus 63%. Different patterns of abnormalities (especially the type, size, and location) were noted in the leukemia subtypes. Copy neutral loss of heterozygosity lesions was detected in 23% of AML‐NK, 3% of CBF‐AML, 25% of MDS, 2% of CML, and 20% of ALL. SNP‐A also provided information on cryptic deletions and a variety of aneuploidies in ALL, while the benefit was minimal in CML. In conclusion, different patterns of abnormal lesions were presented according to the disease category, thus requiring a different approach of adopting SNP‐A‐based karyotyping among different leukemia subtypes. © 2012 Wiley Periodicals, Inc.
The role of elective neck dissection in patients undergoing salvage laryngectomy
Authors: Amit Moran , Hilly Ohad , Leider–Trejo Leonor , Popovtzer Aharon , Gutfeld Orit , Shvero Jacob , Fliss Dan M. , Cohen Jacob T. , Bachar Gideon , Gil Ziv
Published: September 2012

Background

We investigated the risk of neck metastases in patients undergoing salvage total laryngectomy in association with previous radiotherapy.

Methods

The medical records of 42 patients (51 neck specimens) with clinical N0 classification who underwent salvage total laryngectomy in 2 cancer centers were reviewed. Fourteen patients had previous radiotherapy to the central neck and 28 to the central and lateral neck.

Results

Staging before salvage total laryngectomy was similar in both groups. The risk of neck metastases in the central and central/lateral radiation groups was 12% and 18%, respectively (p = .69). Subgroup analysis revealed that 4 of 8 patients initially presenting with clinically N+ had neck metastases before surgery, versus 2 of 26 for those with clinically N0 (p = .015; relative risk [RR] = 4.67). The risk or metastases in the contralateral neck was 0 of 9.

Conclusion

The risk of neck metastases in patients who undergo either central or central/lateral neck radiotherapy is similar. Elective neck dissection seems appropriate in patients undergoing SLR. © 2012 Wiley Periodicals, Inc. Head Neck 35: 1392–1396, 2013
Case of high‐risk epithelioid hemangioendothelioma of the neck
Authors: Matsuzawa Shingo , Kanazawa Takeharu , Yamaguchi Takehiko , Nishino Hiroshi , Kawada Kazumi , Ichimura Keiichi
Published: September 2012

Background

Epithelioid hemangioendotheliomas (EHEs) of the head and neck region are uncommon malignant neoplasms that exhibit various biologic behaviors characteristic of both low‐ and high‐grade malignancy. A subgroup of EHEs identified as “high‐risk” EHEs because of their size and mitotic activity is associated with an unfavorable clinical course and poor prognosis.

Materials and Results

We describe the treatment of the first and, in terms of size, largest case of high‐risk EHE arising from the neck. Despite wide excision, recurrence occurred 9 months after surgery, as had been expected. However, the tumor was found to express both vascular endothelial growth factor (VEGF) and VEGF receptor 2, indicating the potential of anti‐VEGF therapy in the treatment of such cases.

Conclusion

The finding that a high‐risk EHE arising from the head and neck region is characterized by expression of VEGF and its receptor provides further support for the development of targeted molecular therapies. © 2012 Wiley Periodicals, Inc. Head Neck 35: E317–E320, 2013