bluuebunny/crossref_metadata_2025_split huggingface dataset:
Estimation of HbA1c response to sitagliptin by change in glycated albumin level for 2 weeks
Authors: Hamaguchi Tomoya , Koga Masafumi , Murai Jun , Saito Hiroshi , Tamada Daisuke , Kurebayashi Shogo , Katsuno Tomoyuki , Miyagawa Jun‐ichiro , Namba Mitsuyoshi 
Published: October 2011

AbstractAims/Introduction:  Since glycated albumin (GA) reflects shorter‐term (about 2 weeks) control of plasma glucose levels compared with HbA1c, GA is thought to be a useful glycemic control indicator for the early period following commencement of the treatment of diabetes. In this study, we attempted to estimate HbA1c using the change in GA level before and after the first 2 weeks (ΔGA2w) of administration of sitagliptin, a dipeptidyl peptidase‐4 (DPP‐4) inhibitor.Materials and Methods:  The study included 28 patients with type 2 diabetes who were administered sitagliptin at a dose of 50 mg/day for 12 weeks.Results:  At 2 weeks after administration of sitagliptin, GA markedly decreased, while HbA1c had only slightly decreased. A significant positive correlation was observed between the ΔGA2w and the change in HbA1c before and after the first 12 weeks of administration of sitagliptin (ΔHbA1c12w) (R = 0.793, P < 0.0001). The latter was about 0.6 times the former. The estimated HbA1c after 12 weeks of therapy was calculated by adding ΔGA2w × 0.6 to the baseline HbA1c. A significant positive correlation was observed between the estimated HbA1c and the measured HbA1c after 12 weeks (R = 0.735, P < 0.0001) and both were similar levels.Conclusions:  HbA1c in the first 12 weeks after administration of sitagliptin could be estimated from the formula using the ΔGA2w. (J Diabetes Invest, doi: 10.1111/j.2040‐1124.2011.00167.x, 2011)
Dynamic in-situ eutectic formation for topical drug delivery
Authors: Fiala Sarah , Brown Marc B , Jones Stuart A 
Published: October 2011

Abstract

Objectives
The relationship between the solution-state chemistry of eutectic systems and their transmembrane transport characteristics is difficult to define as these mixtures are sensitive to delivery vehicle-induced penetration enhancement. Through in-situ formation of a molten eutectic mixture using highly evaporative sprays this study aimed to gain an understanding of solution-state thermodynamic and chemical properties of eutectic combinations pertinent to transmembrane transport in the absence of a delivery vehicle.

Methods
In-situ molten lidocaine–prilocaine eutectics were formed using a hydroflouroalkane (HFA) propellant. Transport through silicone membranes and human skin in upright Franz diffusion cells was determined using in-house manufactured creams as controls.

Key findings
The application of the two drugs in an HFA spray produced a molten oil even when the melting point of the drug mixture was above the experimental temperature at the membrane surface. In the absence of vehicle effects, molecule presentation to the membrane interface was most effective using a lidocaine-rich mixture of 0.7% w/w lidocaine:prilocaine – 1985.06 ± 128.87 µg/h/cm2.

Conclusions
There appeared to be no link between melting point and transmembrane transport of lidocaine:prilocaine from a eutectic mixture. The rate of drug presentation to the membrane interface, which was highest in drug-rich, high-activity molten eutectic mixtures, was the driver for transmembrane transport in the absence of significant barrier interactions.
Hemin-coupled iron(III)-hydroxide nanoparticles show increased uptake in Caco-2 cells
Authors: Jahn Markus Richard , Shukoor Ibrahim , Tremel Wolfgang , Wolfrum Uwe , Kolb Ute , Nawroth Thomas , Langguth Peter 
Published: October 2011

Abstract

Objectives
The absorption of commonly used ferrous iron salts from intestinal segments at neutral to slightly alkaline pH is low, mainly because soluble ferrous iron is easily oxidized to poorly soluble ferric iron and ferrous iron but not ferric iron is carried by the divalent metal transporter DMT-1. Moreover, ferrous iron frequently causes gastrointestinal side effects. In iron(III)-hydroxide nanoparticles hundreds of ferric iron atoms are safely packed in nanoscaled cores surrounded by a solubilising carbohydrate shell, yet bioavailability from such particles is insufficient when compared with ferrous salts. To increase their intestinal uptake iron(III)-hydroxide nanoparticles were coupled in this study with the protoporphyrin hemin, which undergoes carrier-mediated uptake in the intestine.

Methods
Uptake of iron(III)-hydroxide nanoparticles with hemin covalently coupled by DCC reaction was measured in Caco-2 cells with a colorimetric assay and visualized by transmission electron microscopy.

Key findings
Nanoparticles were taken up by carrier-mediated transport, since uptake was temperature-dependent and increased with an increasing hemin substitution grade. Furthermore, uptake decreased with an increasing concentration of free hemin, due to competition for carrier-mediated uptake.

Conclusions
Hemin-coupled iron(III)-hydroxide nanoparticles were carried by a heme specific transport system, probably via receptor mediated endocytosis. It can be expected that this system shows improved absorption of iron compared with uncoupled iron(III)-hydroxide nanoparticles, which exist on the market today.
Complete Genome Sequence of Strain S85, a Novel Member of the Family Flavobacteriaceae
Authors: Oh Chulhong , Kwon Young-Kyung , Heo Soo-Jin , De Zoysa Mahanama , Affan Abu , Lee Youngdeuk , Lee Jehee , Choi Young-Ung , Park Heung-Sik , Jung Kyung-Hwan , Lee Hyeon-Yong , Kang Do-Hyung 
Published: October 2011

ABSTRACT

An agar-degrading marine bacterium identified as a novel member of the family
Flavobacteriaceae
(strain S85) was isolated from seawater in Micronesia. The sequenced strain S85 genome is composed of 3,384,629 bp in a circular chromosome, which includes 2,883 complete open reading frames.
VPg-Primed RNA Synthesis of Norovirus RNA-Dependent RNA Polymerases by Using a Novel Cell-Based Assay
Authors: Subba-Reddy Chennareddy V. , Goodfellow Ian , Kao C. Cheng 
Published: October 2011

ABSTRACT

Molecular studies of human noroviruses (NoV) have been hampered by the lack of a permissive cell culture system. We have developed a sensitive and reliable mammalian cell-based assay for the human NoV GII.4 strain RNA-dependent RNA polymerase (RdRp). The assay is based on the finding that RNAs synthesized by transiently expressed RdRp can stimulate retinoic acid-inducible gene I (RIG-I)-dependent reporter luciferase production via the beta interferon promoter. Comparable activities were observed for the murine norovirus (MNV) RdRp. RdRps with mutations at divalent metal ion binding residues did not activate RIG-I signaling. Furthermore, both NoV and MNV RdRp activities were stimulated by the coexpression of their respective VPg proteins, while mutations in the putative site of nucleotide linkage on VPg abolished most of their stimulatory effects. Sequencing of the RNAs linked to VPg revealed that the cellular
trans
-Golgi network protein 2 (TGOLN2) mRNA was the template for VPg-primed RNA synthesis. Small interfering RNA knockdown of RNase L abolished the enhancement of signaling that occurred in the presence of VPg. Finally, the coexpression of each of the other NoV proteins revealed that p48 (also known as NS1-2) and VP1 enhanced and that VP2 reduced the RdRp activity. The assay should be useful for the dissection of the requirements for NoV RNA synthesis as well as the identification of inhibitors of the NoV RdRp.
The Structure of an Unconventional HD-GYP Protein from
Bdellovibrio
Reveals the Roles of Conserved Residues in this Class of Cyclic-di-GMP Phosphodiesterases
Authors: Lovering Andrew L. , Capeness Michael J. , Lambert Carey , Hobley Laura , Sockett R. Elizabeth 
Published: October 2011

ABSTRACT

Cyclic-di-GMP is a near-ubiquitous bacterial second messenger that is important in localized signal transmission during the control of various processes, including virulence and switching between planktonic and biofilm-based lifestyles. Cyclic-di-GMP is synthesized by GGDEF diguanylate cyclases and hydrolyzed by EAL or HD-GYP phosphodiesterases, with each functional domain often appended to distinct sensory modules. HD-GYP domain proteins have resisted structural analysis, but here we present the first structural representative of this family (1.28 Å), obtained using the unusual Bd1817 HD-GYP protein from the predatory bacterium
Bdellovibrio bacteriovorus
. Bd1817 lacks the active-site tyrosine present in most HD-GYP family members yet remains an excellent model of their features, sharing 48% sequence similarity with the archetype RpfG. The protein structure is highly modular and thus provides a basis for delineating domain boundaries in other stimulus-dependent homologues. Conserved residues in the HD-GYP family cluster around a binuclear metal center, which is observed complexed to a molecule of phosphate, providing information on the mode of hydroxide ion attack on substrate. The fold and active site of the HD-GYP domain are different from those of EAL proteins, and restricted access to the active-site cleft is indicative of a different mode of activity regulation. The region encompassing the GYP motif has a novel conformation and is surface exposed and available for complexation with binding partners, including GGDEF proteins.

IMPORTANCE
It is becoming apparent that many bacteria use the signaling molecule cyclic-di-GMP to regulate a variety of processes, most notably, transitions between motility and sessility. Importantly, this regulation is central to several traits implicated in chronic disease (adhesion, biofilm formation, and virulence gene expression). The mechanisms of cyclic-di-GMP synthesis via GGDEF enzymes and hydrolysis via EAL enzymes have been suggested by the analysis of several crystal structures, but no information has been available to date for the unrelated HD-GYP class of hydrolases. Here we present the multidomain structure of an unusual member of the HD-GYP family from the predatory bacterium
Bdellovibrio bacteriovorus
and detail the features that distinguish it from the wider structural family of general HD fold hydrolases. The structure reveals how a binuclear iron center is formed from several conserved residues and provides a basis for understanding HD-GYP family sequence requirements for c-di-GMP hydrolysis.
Recombinase-Mediated Cassette Exchange as a Novel Method To Study Somatic Hypermutation in Ramos Cells
Authors: Baughn Linda B. , Kalis Susan L. , MacCarthy Thomas , Wei Lirong , Fan Manxia , Bergman Aviv , Scharff Matthew D. 
Published: October 2011

ABSTRACTActivation-induced cytidine deaminase (AID) mediates the somatic hypermutation (SHM) of immunoglobulin (Ig) variable (V) regions that is required for the generation of antibody diversity and for the affinity maturation of the antibody response against infectious agents and toxic substances. AID preferentially targets WRC (W = A/T, R = A/G) hot spot motifs, particularly WGCW motifs that create overlapping hot spots on both strands. In order to gain a better understanding of the generation of antibody diversity and to create a platform for thein vitrogeneration of affinity-matured antibodies, we have established a system involving recombinase-mediated cassette exchange (RMCE) to replace the V region and its flanking sequences. This makes it possible to easily manipulate the sequence of the Ig gene within the endogenous heavy chain of the Ramos human Burkitt’s lymphoma cell line. Here we show that the newly integrated wild-type (WT) VH regions introduced by RMCE undergo SHM similarly to non-RMCE-modified Ramos cells. Most importantly, we have shown that introducing a cluster of WGCW motifs into the complementary determining region 2 (CDR2) of the human heavy chain V region significantly raised the mutation frequency and number of mutations per sequence compared to WT controls. Thus, we have demonstrated a novel platform in Ramos cells whereby we can easily and quickly manipulate the endogenous human VH region to further explore the regulation and targeting of SHM. This platform will be useful for generating human antibodies with changes in affinity and specificityin vitro.IMPORTANCEAn effective immune response requires a highly diverse repertoire of affinity-matured antibodies. Activation-induced cytidine deaminase (AID) is required for somatic hypermutation (SHM) of immunoglobulin (Ig) genes. Although a great deal has been learned about the regulation of AID, it remains unclear how it is preferentially targeted to particular motifs, to certain locations within the Ig gene and not to other highly expressed genes in the germinal center B cell. This is an important question because AID is highly mutagenic and is sometimes mistargeted to other highly expressed genes, including proto-oncogenes, leading to B cell lymphomas. Here we describe how we utilize recombinase-mediated cassette exchange (RMCE) to modify the sequence of the endogenous heavy chain locus in the Ramos Burkitt’s lymphoma cell line. This platform can be used to explore the regulation and targeting of SHM and to generate human antibodies with changes in affinity and specificityin vitro.
Mucin Glycosylating Enzyme GALNT2 Regulates the Malignant Character of Hepatocellular Carcinoma by Modifying the EGF Receptor
Authors: Wu Yao-Ming , Liu Chiung-Hui , Hu Rey-Heng , Huang Miao-Juei , Lee Jian-Jr , Chen Chi-Hau , Huang John , Lai Hong-Shiee , Lee Po-Huang , Hsu Wen-Ming , Huang Hsiu-Chin , Huang Min-Chuan 
Published: October 2011

Abstract
Extracellular glycosylation is a critical determinant of malignant character. Here, we report that N-acetylgalactosaminyltransferase 2 (GALNT2), the enzyme that mediates the initial step of mucin type-O glycosylation, is a critical mediator of malignant character in hepatocellular carcinoma (HCC) that acts by modifying the activity of the epidermal growth factor receptor (EGFR). GALNT2 mRNA and protein were downregulated frequently in HCC tumors where these events were associated with vascular invasion and recurrence. Restoring GALNT2 expression in HCC cells suppressed EGF-induced cell growth, migration, and invasion in vitro and in vivo. Mechanistic investigations revealed that the status of the O-glycans attached to the EGFR was altered by GALNT2, changing EGFR responses after EGF binding. Inhibiting EGFR activity with erlotinib decreased the malignant characters caused by siRNA-mediated knockdown of GALNT2 in HCC cells, establishing the critical role of EGFR in mediating the effects of GALNT2 expression. Taken together, our results suggest that GALNT2 dysregulation contributes to the malignant behavior of HCC cells, and they provide novel insights into the significance of O-glycosylation in EGFR activity and HCC pathogenesis. Cancer Res; 71(23); 7270–9. ©2011 AACR.
Dishevelled 2 Signaling Promotes Self-Renewal and Tumorigenicity in Human Gliomas
Authors: Pulvirenti Teodoro , Van Der Heijden Maartje , Droms Leif A. , Huse Jason T. , Tabar Viviane , Hall Alan 
Published: October 2011

Abstract
Glioblastoma multiforme is the most common glioma variant in adults and is highly malignant. Tumors are thought to harbor a subpopulation of stem-like cancer cells, with the bulk resembling neural progenitor-like cells that are unable to fully differentiate. Although multiple pathways are known to be involved in glioma tumorigenesis, the role of Wnt signaling has been poorly described. Here, we show that Dishevelled 2 (Dvl2), a key component of the Wnt signaling pathway, is overexpressed in human gliomas. RNA interference–mediated depletion of Dvl2 blocked proliferation and promoted the differentiation of cultured human glioma cell lines and primary, patient-derived glioma cells. In addition, Dvl2 depletion inhibited tumor formation after intracranial injection of glioblastoma cells in immunodeficient mice. Inhibition of canonical Wnt/β-catenin signaling also blocked proliferation, but unlike Dvl2 depletion, did not induce differentiation. Finally, Wnt5a, a noncanonical Wnt ligand, was also required for glioma cell proliferation. The data therefore suggest that both canonical and noncanonical Wnt signaling pathways downstream of Dvl2 cooperate to maintain the proliferative capacity of human glioblastomas. Cancer Res; 71(23); 7280–90. ©2011 AACR.
Disentangling the Myeloma Web
Authors: Tonon Giovanni , Anderson Kenneth C. 
Published: October 2011

Abstract
Gene expression profiling in patients with multiple myeloma has identified gene signatures linked to prognosis. However, because of their heterogeneity, there is currently no consensus about which signatures represent the best predictive and prognostic markers. Novel computational tools are now helping investigators integrate heterogeneous data sets to identify universal and robust classifiers. Clin Cancer Res; 17(23); 7210–2. ©2011 AACR.
Quiescin Sulfhydryl Oxidase 1 Promotes Invasion of Pancreatic Tumor Cells Mediated by Matrix Metalloproteinases
Authors: Katchman Benjamin A. , Antwi Kwasi , Hostetter Galen , Demeure Michael J. , Watanabe Aprill , Decker G. Anton , Miller Laurence J. , Von Hoff Daniel D. , Lake Douglas F. 
Published: October 2011

Abstract
Quiescin sulfhydryl oxidase 1 (QSOX1) oxidizes sulfhydryl groups to form disulfide bonds in proteins. We previously mapped a peptide in plasma from pancreatic ductal adenocarcinoma (PDA) patients back to an overexpressed QSOX1 parent protein. In addition to overexpression in pancreatic cancer cell lines, 29 of 37 patients diagnosed with PDA expressed QSOX1 protein in tumor cells, but QSOX1 was not detected in normal adjacent tissues or in a transformed, but nontumorigenic cell line. To begin to evaluate the advantage QSOX1 might provide to tumors, we suppressed QSOX1 protein expression using short hairpin (sh) RNA in two pancreatic cancer cell lines. Growth, cell cycle, apoptosis, invasion, and matrix metalloproteinase (MMP) activity were evaluated. QSOX1 shRNA suppressed both short and long isoforms of the protein, showing a significant effect on cell growth, cell cycle, and apoptosis. However, QSOX1 shRNA dramatically inhibited the abilities of BxPC-3 and Panc-1 pancreatic tumor cells to invade through Matrigel in a modified Boyden chamber assay. Mechanistically, gelatin zymography indicated that QSOX1 plays an important role in activation of MMP-2 and MMP-9. Taken together, our results suggest that the mechanism of QSOX1-mediated tumor cell invasion is by activation of MMP-2 and MMP-9. Mol Cancer Res; 9(12); 1621–31. ©2011 AACR.
Use of Early Clopidogrel by Reperfusion Strategy Among Patients Presenting With ST-Segment Elevation Myocardial Infarction
Authors: Melloni Chiara , Roe Matthew T. , Chen Anita Y. , Wang Tracy Y. , Wiviott Stephen D. , Ho P. Michael , Peterson Eric D. , Alexander Karen P. 
Published: October 2011

Background—
The 2007 update of the ACC/AHA guidelines for STEMI patients recommended addition of clopidogrel to aspirin regardless of reperfusion strategy, with a bolus dose in patients <75 years of age.

Methods and Results—
We evaluated use and dose of early clopidogrel among 52,140 STEMI patients enrolled in 368 hospitals participating in NCDR's ACTION Registry®–Get with the Guidelines (GWTG™) from January 2007–September 2009. Patients were stratified by reperfusion strategy: primary percutaneous coronary intervention (PCI, n=37,108), fibrinolysis (n=5805), or no-reperfusion (n=9227), and by age (<75 or ≥75 years). Adjusted odds for in-hospital outcomes are reported by clopidogrel use across reperfusion strategies. Clopidogrel was administered early to 97% of primary PCI, 18% of fibrinolytic, and 6% of non-reperfused patients. Among patients receiving clopidogrel, a loading dose (≥300 mg) was often used in primary PCI (91%) but less frequently among fibrinolysis-treated (83%) and non-reperfused patients (74%). A positive time trend from Q1 2007–Q3 2009 in overall clopidogrel use was observed only in fibrinolytic patients (15–20%) Use of clopidogrel was associated with a significant increase in major bleeding only among older patients in the no-reperfusion group (21.9% vs. 13.2%; OR 2.19; 95% CI 1.47–3.27). A significantly lower risk of in-hospital death was associated with clopidogrel use across all reperfusion strategies (OR [95% CI], primary PCI: 0.15 [0.13–0.19]; fibrinolysis: 0.26 [0.12–0.57]; no reperfusion: 0.42 [0.27–0.65]).

Conclusion—
Early clopidogrel use has not yet extended to the routine care of STEMI patients treated with fibrinolysis or those not receiving reperfusion as recommended in the guideline update.
Similarities and Differences of Diagnostic Manual Tests for Anterior Cruciate Ligament Insufficiency
Authors: Kuroda Ryosuke , Hoshino Yuichi , Kubo Seiji , Araki Daisuke , Oka Shinya , Nagamune Kouki , Kurosaka Masahiro 
Published: October 2011

Background: The Lachman and pivot-shift tests are 2 standard manual tests to diagnose anterior cruciate ligament (ACL) insufficiency. However, the global variation of these testing procedures is not known. Purpose: To survey currently used testing techniques and to measure the knee movement during manual tests among various expert surgeons from across the globe. Study Design: Controlled laboratory study. Methods: Part 1: descriptive survey. A questionnaire asking about testing procedures of Lachman and pivot-shift tests was conducted among 33 ACL surgeons. Part 2: knee kinematics comparison. Lachman and pivot-shift tests were performed on a unilateral ACL-injured patient by 5 surgeons, while knee kinematics was recorded by an electromagnetic system. Tibial translation was measured during the Lachman test, while tibial translation, rotation, and pivot-shift acceleration were calculated during the pivot-shift test. Results: Part 1: Tibial anterior drawer by a medially placed hand was widely advocated for the Lachman test. Flexion type of the pivot-shift test maneuver was supported by two thirds, while extension type was supported by one third. However, the “feeling” of subluxation or reduction during the pivot shift was the primary evaluation method used by the vast majority of surgeons. Part 2: Increased tibial translation during the Lachman test was observed in the ACL-injured knee with significant variation between examiners ( P < .01). Tibial translation and pivot-shift acceleration during the pivot-shift test increased in the ACL-injured side ( P < .01), but tibial rotation was too diverse to find any trend ( P = .31). Tibial translation and acceleration of the pivot shift in the ACL-injured knee showed no significant difference between examiners ( P > .05). Conclusion: The Lachman test can display a wide variation of actual movement despite maneuver similarity, while the pivot-shift test could possibly be measurable by tibial translation and/or acceleration beyond their procedural variation. Clinical Relevance: We should recognize the limitations of these manual tests and the possibilities of their objective measurement.
Somatic mutations in the transcriptional corepressor gene BCORL1 in adult acute myelogenous leukemia
Authors: Li Meng , Collins Roxane , Jiao Yuchen , Ouillette Peter , Bixby Dale , Erba Harry , Vogelstein Bert , Kinzler Kenneth W. , Papadopoulos Nickolas , Malek Sami N. 
Published: October 2011

Abstract
To further our understanding of the genetic basis of acute myelogenous leukemia (AML), we determined the coding exon sequences of ∼ 18 000 protein-encoding genes in 8 patients with secondary AML. Here we report the discovery of novel somatic mutations in the transcriptional corepressor gene BCORL1 that is located on the X-chromosome. Analysis of BCORL1 in an unselected cohort of 173 AML patients identified a total of 10 mutated cases (6%) with BCORL1 mutations, whereas analysis of 19 AML cell lines uncovered 4 (21%) BCORL1 mutated cell lines. The majority (87%) of the mutations in BCORL1 were predicted to inactivate the gene product as a result of nonsense mutations, splice site mutation, or out-of-frame insertions or deletions. These results indicate that BCORL1 by genetic criteria is a novel candidate tumor suppressor gene, joining the growing list of genes recurrently mutated in AML.