bluuebunny/crossref_metadata_2025 huggingface dataset:
Application of microchip CGE for the analysis of PEG‐modified recombinant human granulocyte‐colony stimulating factors
Authors: Park Eun Ji , Lee Kyung Soo , Lee Kang Choon , Na Dong Hee
Published: October 2010

The purpose of this study was to evaluate the microchip CGE (MCGE) for the analysis of PEG‐modified granulocyte‐colony stimulating factor (PEG‐G‐CSF) prepared with PEG‐aldehydes. The unmodified and PEG‐modified G‐CSFs were analyzed by Protein 80 and 230 Labchips on the Agilent 2100 Bioanalyzer. The MCGE allowed size‐based separation and quantitation of PEG‐G‐CSF. The Protein 80 Labchip was useful for PEG‐5K‐G‐CSF, while the Protein 230 Labchip was more suitable for PEG‐20K‐G‐CSF. The MCGE was also used to monitor a search for optimal PEG‐modification (PEGylation) conditions to produce mono‐PEG‐G‐CSF. This study demonstrates the usefulness of MCGE for monitoring and optimizing the PEGylation of G‐CSF with the advantages of speed, minimal sample consumption, and automatic quantitation.
Consistent MYC and FLT4 gene amplification in radiation‐induced angiosarcoma but not in other radiation‐associated atypical vascular lesions
Authors: Guo Tianhua , Zhang Lei , Chang Ning‐En , Singer Samuel , Maki Robert G. , Antonescu Cristina R
Published: October 2010

Angiosarcoma (AS) is a distinct group of sarcomas characterized by upregulation of vascular‐specific receptor tyrosine kinases, including TIE1, KDR, TEK, and FLT1. In keeping with the clinical heterogeneity, gene‐expression profiling distinguishes two AS genomic clusters, which correlate with anatomical location and prior exposure to radiation. Furthermore, a high percentage of secondary AS, but not primary AS, shows distinct 8q24 chromosomal gains, due to MYC amplification. In this study, we mined the transcriptional output of 10 secondary and 11 primary AS to better define the dichotomy in the pathogenesis of these two clinical subsets. The oncogenic role of MYC was investigated further in secondary AS as well as in radiation‐induced atypical vascular lesions (AVL) and other radiation‐associated sarcomas. High‐level MYC amplification was found in 100% of secondary AS, but in none of the AVL or other radiation‐associated sarcomas. Coamplification of FLT4 (encoding VEGFR3) was identified in 25% of secondary AS, but not in other types. Our findings reinforce the distinct pathogenesis of AS subtypes, with MYC amplification being an early, but necessary event in secondary AS. Secondary genetic hits, such as FLT4 gene coamplification or KDR mutations, may play a role in tumor progression as well as potential therapeutic targeting. © 2010 Wiley‐Liss, Inc.
Synthesis and antifungal activity of novel 5‐substituted 4‐(1,3,4‐thiadiazol‐2‐yl)benzene‐1,3‐diols
Authors: Matysiak Joanna , Skrzypek Alicja , Niewiadomy Andrzej
Published: October 2010

5‐Substituted (amine, alkyl, aryl, heterocyclic) 4‐(1,3,4‐thiadiazol‐2‐yl)benzene‐1,3‐ diols were synthesized, and their antifungal properties were examined. The compounds were obtained by the one‐pot reaction of sulfinylbis((2,4‐dihydroxyphenyl)methanethione) with hydrazides or thiosemicarbazides. Their structures were identified from elemental, IR, 1H NMR, and MS spectra analyses. The activities of the derivatives against five phytopathogenic fungi in vitro were measured. Moderate fungicidal effect of the compounds under consideration was found. © 2010 Wiley Periodicals, Inc. Heteroatom Chem 21:533–540, 2010; View this article online at wileyonlinelibrary.com. DOI 10.1002/hc.20645
Higher bone mineral density is associated with a decreased risk of colorectal adenomas
Authors: Nock Nora L. , Patrick‐Melin Aimee , Cook Marc , Thompson Cheryl , Kirwan John P. , Li Li
Published: October 2010

Bone mineral density (BMD) is a biomarker for cumulative exposure to multiple factors including estrogen, calcium, vitamin D and physical activity, which have all been independently associated with colorectal cancer. Furthermore, higher levels of BMD have been inversely associated with colorectal cancer risk, particularly in postmenopausal women. However, no prior studies have examined the potential association between BMD and colorectal adenomas, which are precursor lesions to most colorectal cancers. Therefore, we evaluated the association between BMD, which was measured using a whole body, dual‐energy X‐ray absorptiometry scan and colorectal adenomas in 167 patients who underwent colonoscopy screening. We found that patients in the highest tertile of total body BMD (>1.294 g/cm2) and in the middle tertile (≥1.167 to ≤1.294 g/cm2) compared to those with a total body BMD in the lowest tertile (<1.167 g/cm2) had a lower risk of colorectal adenomas (highest vs. lowest tertile: OR = 0.29 (0.10–0.84); middle vs. lowest tertile: OR = 0.26 (0.08–0.80); p‐trend = 0.02). Stratification by gender revealed that this association was more pronounced in women (highest (>1.280 g/cm2) vs. lowest (<1.130 g/cm2) tertile: OR = 0.08 (0.01–0.70); middle (≥1.130 to ≤1.280 g/cm2) vs. lowest tertile: OR = 0.15 (0.04–0.94); p‐trend = 0.02) even after excluding hormone replacement therapy users (highest (>1.295 g/cm2) and middle (≥1.132 to ≤1.295 g/cm2) vs. lowest (<1.132 g/cm2) tertile: OR = 0.17 (0.03–0.97); p‐trend = 0.04). Our results show, for the first time, that BMD is inversely associated with colorectal adenomas, particularly in women. Although additional larger, prospective studies are needed, our results suggest that BMD may be a biomarker for colorectal cancer precursor lesions.
Modelling dependence using skew t copulas: Bayesian inference and applications
Authors: Smith Michael S. , Gan Quan , Kohn Robert J.
Published: October 2010

We construct a copula from the skew t distribution of Sahu et al. (2003). This copula can capture asymmetric and extreme dependence between variables, and is one of the few copulas that can do so and still be used in high dimensions effectively. However, it is difficult to estimate the copula model by maximum likelihood when the multivariate dimension is high, or when some or all of the marginal distributions are discrete‐valued, or when the parameters in the marginal distributions and copula are estimated jointly. We therefore propose a Bayesian approach that overcomes all these problems. The computations are undertaken using a Markov chain Monte Carlo simulation method which exploits the conditionally Gaussian representation of the skew t distribution. We employ the approach in two contemporary econometric studies. The first is the modelling of regional spot prices in the Australian electricity market. Here, we observe complex non‐Gaussian margins and nonlinear inter‐regional dependence. Accurate characterization of this dependence is important for the study of market integration and risk management purposes. The second is the modelling of ordinal exposure measures for 15 major websites. Dependence between websites is important when measuring the impact of multi‐site advertising campaigns. In both cases the skew t copula substantially outperforms symmetric elliptical copula alternatives, demonstrating that the skew t copula is a powerful modelling tool when coupled with Bayesian inference. Copyright © 2010 John Wiley & Sons, Ltd.
Reprogramming the pluripotent cell cycle: Restoration of an abbreviated G1 phase in human induced pluripotent stem (iPS) cells
Authors: Ghule Prachi N. , Medina Ricardo , Lengner Christopher J. , Mandeville Matthew , Qiao Meng , Dominski Zbigniew , Lian Jane B. , Stein Janet L. , van Wijnen Andre J. , Stein Gary S.
Published: October 2010

Induced pluripotent stem (iPS) cells derived from terminally differentiated human fibroblasts are reprogrammed to possess stem cell like properties. However, the extent to which iPS cells exhibit unique properties of the human embryonic stem (hES) cell cycle remains to be established. hES cells are characterized by an abbreviated G1 phase (∼2.5 h) and accelerated organization of subnuclear domains that mediate the assembly of regulatory machinery for histone gene expression [i.e., histone locus bodies (HLBs)]. We therefore examined cell cycle parameters of iPS cells in comparison to hES cells. Analysis of DNA synthesis [5‐bromo‐2′‐deoxy‐uridine (BrdU) incorporation], cell cycle distribution (FACS analysis and Ki67 staining) and subnuclear organization of HLBs [immunofluorescence microscopy and fluorescence in situ hybridization (FISH)] revealed that human iPS cells have a short G1 phase (∼2.5 h) and an abbreviated cell cycle (16–18 h). Furthermore, HLBs are formed and reorganized rapidly after mitosis (within 1.5–2 h). Thus, reprogrammed iPS cells have cell cycle kinetics and dynamic subnuclear organization of regulatory machinery that are principal properties of pluripotent hES cells. Our findings support the concept that the abbreviated cell cycle of hES and iPS cells is functionally linked to pluripotency. J. Cell. Physiol. 226: 1149–1156, 2011. © 2010 Wiley‐Liss, Inc.
Oncoprotein Cot1 represses kinase suppressors of Ras1/2 and 1,25‐dihydroxyvitamin D3‐induced differentiation of human acute myeloid leukemia cells
Authors: Wang Xuening , Studzinski George P.
Published: October 2010

Metabolites and derivatives of vitamin D are well‐known inducers of monocytic differentiation, but the mechanistic basis for their action is not fully elucidated. Here we show that the product of protooncogene Cot1 represses the monocytic phenotype in human acute myeloid leukemia (AML) cells induced to differentiate by 1,25‐dihydroxyvitamin D3 (1,25D), even though the expression of cellular Cot1 increases early in the process of 1,25D‐induced differentiation. Interestingly, the expression of the two members of the Kinase Suppressor of Ras (KSR) family of molecular scaffolds, known to be positive regulators of Ras signaling and of 1,25D‐induced differentiation, increases in parallel with Cot1 in 1,25D‐treated cells. However, KSR1/2 are negatively regulated by Cot1, as determined by transfection of siCot1, and confirmed by a reverse effect of ectopic expression of Cot1. The effect of Cot1 in AML cells appears to be cell‐type specific, as previous reports in other cell types found KSR‐2 to be a negative regulator of Cot1, a reverse relationship. Also in contrast to findings in other cells, in AML cells Cot1 exerts negative control on the MAP kinase pathways, since siCot1 increases the levels of activated Raf1, p90RSK, JNK1, c‐jun, and p38, though not of MEK/ERK. These findings have implications for therapy of AML, since in AML cells active MAPKs hasten cell differentiation, and specific pharmacological inhibitors of Cot1 kinase activity have recently became available, thus making Cot1 a “druggable” target. J. Cell. Physiol. 226: 1232–1240, 2011. © 2010 Wiley‐Liss, Inc.
PET translates neurophysiology into images: A review to stimulate a network between neuroimaging and basic research
Authors: Giovacchini Giampiero , Squitieri Ferdinando , Esmaeilzadeh Mouna , Milano Amalia , Mansi Luigi , Ciarmiello Andrea
Published: October 2010

In the last decades there has been a progressive advance in the development of techniques able to explore in humans neurophysiologic and neurochemical processes. Positron emission tomography (PET) is a very powerful technique allowing to study a quite variable range of physiological and biochemical processes in the healthy subjects and in diseases. Apart from its capacity to provide pathophysiological information, PET is also important for the objective assessment of therapeutic efficacy. Initial studies were performed measuring cerebral metabolic rate for glucose (CMRglc) and cerebral blood flow (CBF), representing an indirect index of synaptic activity. The advent of receptor tracers allowed measuring other important physiological parameters, such as receptor occupancy, and endogenous release. In neuropsychiatric disorders, as Alzheimer disease, schizophrenia, epilepsy and Huntington disease, PET has been useful to elaborate hypothesis of the pathogenesis, to relate symptoms to biological variables and to study individuals at increased risk. The new concepts of neurovascular unit and default network, preferentially active at rest, can significantly change the approach of PET, with images reflecting a complex scenario, not merely limited to neural activity, but involving the activity of the entire neurovascular unit and the multifunctional role of astrocytes. To detect dysfunction of the dialog between glutamatergic neurons and astrocytes could lead to a better understanding of altered functional brain images. In this direction a professional network between PET researchers and basic scientists, could give a determinant improvement in the capability to understand the complex physiological and pathophysiological cerebral world. J. Cell. Physiol. 226: 948–961, 2011. © 2010 Wiley‐Liss, Inc.
Toll‐like receptors expressed by dermal fibroblasts contribute to hypertrophic scarring
Authors: Wang JianFei , Hori Keijiro , Ding Jie , Huang Yue , Kwan Peter , Ladak Adil , Tredget Edward E.
Published: October 2010

Hypertrophic scar (HTS), a fibroproliferative disorder (FPD), complicates burn wound healing. Although the pathogenesis is not understood, prolonged inflammation is a known contributing factor. Emerging evidence suggests that fibroblasts regulate immune/inflammatory responses through toll‐like receptor 4 (TLR4) activated by lipopolysaccharide (LPS) through adaptor molecules, leading to nuclear factor kappa‐light‐chain‐enhancer of activated B cells and mitogen‐activated protein kinases activation, cytokine gene transcription and co‐stimulatory molecule expression resulting in inflammation. This study explored the possible role of TLR4 in HTS formation. Paired normal and HTS tissue from burn patients was collected and dermal fibroblasts isolated and cultured. Immunohistochemical analysis of tissues demonstrated increased TLR4 staining in HTS tissue. Quantitative RT‐PCR of three pairs of fibroblasts demonstrated mRNA levels for TLR4 and its legend myeloid differentiation factor 88 (MyD88) in HTS fibroblasts were increased significantly compared with normal fibroblasts. Flow cytometry showed increased TLR4 expression in HTS fibroblasts compared with normal. ELISA demonstrated protein levels for prostaglandin E2, interleukin (IL)‐6, IL‐8 and monocyte chemotactic protein‐1 (MCP‐1) were significantly increased in HTS fibroblasts compared to normal. When paired normal and HTS fibroblasts were stimulated with LPS, significant increases in mRNA and protein levels for MyD88, IL‐6, IL‐8, and MCP‐1 were detected. However, when transfected with MyD88 small interfering RNA (siRNA), then stimulated with LPS, a significant decrease in mRNA and protein levels for these molecules compared to only LPS‐stimulated fibroblasts was detected. In comparison, a scramble siRNA transfection did not affect mRNA or protein levels for these molecules. Results demonstrate LPS stimulates proinflammatory cytokine expression in dermal fibroblasts and MyD88 siRNA eliminates the expression. Therefore, controlling inflammation and manipulating TLR signaling in skin cells may result in novel treatment strategies for HTS and other FPD. J. Cell. Physiol. 226: 1265–1273, 2011. © 2010 Wiley‐Liss, Inc.
Protein kinase Cι expression and oncogenic signaling mechanisms in cancer
Authors: Murray Nicole R. , Kalari Krishna R. , Fields Alan P.
Published: October 2010

Accumulating evidence demonstrates that PKCι is an oncogene and prognostic marker that is frequently targeted for genetic alteration in many major forms of human cancer. Functional data demonstrate that PKCι is required for the transformed phenotype of lung, pancreatic, ovarian, prostate, colon, and brain cancer cells. Future studies will be required to determine whether PKCι is also an oncogene in the many other cancer types that also overexpress PKCι. Studies of PKCι using genetically defined models of tumorigenesis have revealed a critical role for PKCι in multiple stages of tumorigenesis, including tumor initiation, progression, and metastasis. Recent studies in a genetic model of lung adenocarcinoma suggest a role for PKCι in transformation of lung cancer stem cells. These studies have important implications for the therapeutic use of aurothiomalate (ATM), a highly selective PKCι signaling inhibitor currently undergoing clinical evaluation. Significant progress has been made in determining the molecular mechanisms by which PKCι drives the transformed phenotype, particularly the central role played by the oncogenic PKCι‐Par6 complex in transformed growth and invasion, and of several PKCι‐dependent survival pathways in chemo‐resistance. Future studies will be required to determine the composition and dynamics of the PKCι‐Par6 complex, and the mechanisms by which oncogenic signaling through this complex is regulated. Likewise, a better understanding of the critical downstream effectors of PKCι in various human tumor types holds promise for identifying novel prognostic and surrogate markers of oncogenic PKCι activity that may be clinically useful in ongoing clinical trials of ATM. J. Cell. Physiol. 226: 879–887, 2011. © 2010 Wiley‐Liss, Inc.
Live cell imaging of the cancer‐related transcription factor RUNX2 during mitotic progression
Authors: Pockwinse Shirwin M. , Kota Krishna P. , Quaresma Alexandre J.C. , Imbalzano Anthony N. , Lian Jane B. , van Wijnen Andre J. , Stein Janet L. , Stein Gary S. , Nickerson Jeffrey A.
Published: October 2010

The nuclear matrix bound transcription factor RUNX2 is a lineage‐specific developmental regulator that is linked to cancer. We have previously shown that RUNX2 controls transcription of both RNA polymerase II genes and RNA polymerase I‐dependent ribosomal RNA genes. RUNX2 is epigenetically retained through mitosis on both classes of target genes in condensed chromosomes. We have used fluorescence recovery after photobleaching to measure the relative binding kinetics of enhanced green fluorescent protein (EGFP)‐RUNX2 at transcription sites in the nucleus and nucleoli during interphase, as well as on mitotic chromosomes. RUNX2 becomes more strongly bound as cells go from interphase through prophase, with a doubling of the most tightly bound “immobile fraction.” RUNX2 exchange then becomes much more facile during metaphase to telophase. During interphase the less tightly bound pool of RUNX2 exchanges more slowly at nucleoli than at subnuclear foci, and the non‐exchanging immobile fraction is greater in nucleoli. These results are consistent with a model in which the molecular mechanism of RUNX2 binding is different at protein‐coding and ribosomal RNA genes. The binding interactions of RUNX2 change as cells go through mitosis, with binding affinity increasing as chromosomes condense and then decreasing through subsequent mitotic phases. The increased binding affinity of RUNX2 at mitotic chromosomes may reflect its epigenetic function in “bookmarking” of target genes in cancer cells. J. Cell. Physiol. 226: 1383–1389, 2011. © 2010 Wiley‐Liss, Inc.
5‐hydroxy‐2‐(4‐hydroxy‐3‐methoxyphenyl)‐3,7‐dimethoxy‐4H‐chromen‐4‐one (MSF‐2) suppresses fMLP‐mediated respiratory burst in human neutrophils by inhibiting phosphatidylinositol 3‐kinase activity
Authors: Liao Chang‐Hui , Chen Jih‐Jung , Lin Jieru Egeria , Liu Chia‐Hsin , Tseng Ching‐Ping , Day Yuan‐Ji
Published: October 2010

Respiratory burst mediates crucial bactericidal mechanism in neutrophils. However, undesirable respiratory burst leads to pathological inflammation and tissue damage. This study investigates the effect and the underlying mechanism of 5‐hydroxy‐2‐(4‐hydroxy‐3‐methoxyphenyl)‐3,7‐dimethoxy‐4H‐chromen‐4‐one (MSF‐2), a lignan extracted from the fruit of Melicope Semecarprifolia, on fMLP‐induced respiratory burst in human neutrophils and suggests a possible therapeutic approach to ameliorate disease associated with neutrophil hyperactivation. MSF‐2 inhibited fMLP‐induced neutrophil superoxide anion production, cathepsin G release and migration in human neutrophils isolated from healthy volunteers, reflecting inhibition of phosphatidylinositol 3‐kinase (PI3K) activation. Specifically, PI3K/AKT activation results in migration, degranulation and superoxide anion production in neutrophils. MSF‐2 suppresses PI3K activation and phosphatidylinositol (3,4,5)‐trisphosphate (PIP3) production, and consequently inhibits downstream activation of PDK1 and AKT. Further, PI3K also stimulates respiratory burst via PLC‐dependent elevation of intracellular calcium. MSF‐2 reduces fMLP‐mediated PLCγ2 activation and intracellular calcium accumulation notably through extracellular calcium influx in a PI3K and PLC‐dependent manner. However, MSF‐2 is not a competitive or allosteric antagonist of fMLP. Additionally, in an in vivo study, MSF‐2 prevents fMLP‐induced neutrophil infiltration and inflammation in mice. In conclusion, MSF‐2 opposes fMLP‐mediated neutrophil activation and inflammation by inhibiting PI3K activation and subsequent activation of AKT and PLCγ2. J. Cell. Physiol. 226: 1519–1530, 2011. © 2010 Wiley‐Liss, Inc.
FTS (fused toes homolog) a novel oncoprotein involved in uterine cervical carcinogenesis and a potential diagnostic marker for cervical cancer
Authors: Cinghu Senthilkumar , Anandharaj Arunkumar , Lee Ho‐chang , Yu Jae‐Ran , Park Woo‐Yoon
Published: October 2010

The high incidence and fatality rate of uterine cervical cancer warrant effective diagnostic and therapeutic target identification for this disease. Here, we have found a novel oncoprotein FTS (Fused Toes Homolog), which is involved in cervical cancer pathogenesis. Immnohistochemical analysis of human cervical biopsy samples revealed that the expression of FTS is absent in normal cervical epithelium but progressively overexpressed in human cervical intraneoplastic lesions (CIN‐I to CIN‐III), this characteristic phenomenon put this protein, a potential diagnostic marker for the screening of early neoplastic changes of cervix. Using FTS‐specific small hairpin RNA (shRNA) in cervical cancer cells, we determined a specific role for FTS protein in, cervical neoplasia. Targeted stable knock down of FTS in HeLa cells led to the growth inhibition, cell‐cycle arrest, and apoptosis with concurrent increase in p21 protein. FTS effectively represses the p21 mRNA expression in dual luciferase assay which indicates that p21 is transcriptionally regulated by this oncoprotein which in turn affect the regular cell‐cycle process and its components. Consistent with this we found a reciprocal association between these proteins in early cervical neoplastic tissues. These data unraveled the involvement of new oncoprotein FTS in cervical cancer which plays a central role in carcinogenesis. Targeted inhibition of FTS lead to the shutdown of key elemental characteristics of cervical cancer and could lead to an effective therapeutic strategy for cervical cancer. J. Cell. Physiol. 226: 1564–1572, 2011. © 2010 Wiley‐Liss, Inc.
The role of electrical signals in murine corneal wound re‐epithelialization
Authors: Kucerova Romana , Walczysko Petr , Reid Brian , Ou Jingxing , Leiper Lucy J. , Rajnicek Ann M. , McCaig Colin D. , Zhao Min , Collinson J Martin
Published: October 2010

Ion flow from intact tissue into epithelial wound sites results in lateral electric currents that may represent a major driver of wound healing cell migration. Use of applied electric fields (EF) to promote wound healing is the basis of Medicare‐approved electric stimulation therapy. This study investigated the roles for EFs in wound re‐epithelialization, using the Pax6+/− mouse model of the human ocular surface abnormality aniridic keratopathy (in which wound healing and corneal epithelial cell migration are disrupted). Both wild‐type (WT) and Pax6+/− corneal epithelial cells showed increased migration speeds in response to applied EFs in vitro. However, only Pax6+/+ cells demonstrated consistent directional galvanotaxis towards the cathode, with activation of pSrc signaling, polarized to the leading edges of cells. In vivo, the epithelial wound site normally represents a cathode, but 43% of Pax6+/− corneas exhibited reversed endogenous wound‐induced currents (the wound was an anode). These corneas healed at the same rate as WT. Surprisingly, epithelial migration did not correlate with direction or magnitude of endogenous currents for WT or mutant corneas. Furthermore, during healing in vivo, no polarization of pSrc was observed. We found little evidence that Src‐dependent mechanisms of cell migration, observed in response to applied EFs in vitro, normally exist in vivo. It is concluded that endogenous EFs do not drive long‐term directionality of sustained healing migration in this mouse corneal epithelial model. Ion flow from wounds may nevertheless represent an important component of wound signaling initiation. J. Cell. Physiol. 226: 1544–1553, 2011. © 2010 Wiley‐Liss, Inc.
Potential role of follicle‐stimulating hormone (FSH) and transforming growth factor (TGFβ1) in the regulation of ovarian angiogenesis
Authors: Kuo Shih‐Wei , Ke Ferng‐Chun , Chang Geen‐Dong , Lee Ming‐Ting , Hwang Jiuan‐Jiuan
Published: October 2010

Angiogenesis occurs during ovarian follicle development and luteinization. Pituitary secreted FSH was reported to stimulate the expression of endothelial mitogen VEGF in granulosa cells. And, intraovarian cytokine transforming growth factor (TGF)β1 is known to facilitate FSH‐induced differentiation of ovarian granulosa cells. This intrigues us to investigate the potential role of FSH and TGFβ1 regulation of granulosa cell function in relation to ovarian angiogenesis. Granulosa cells were isolated from gonadotropin‐primed immature rats and treated once with FSH and/or TGFβ1 for 48 h, and the angiogenic potential of conditioned media (granulosa cell culture conditioned media; GCCM) was determined using an in vitro assay with aortic ring embedded in collagen gel and immunoblotting. FSH and TGFβ1 increased the secreted angiogenic activity in granulosa cells (FSH + TGFβ1 > FSH ≈ TGFβ1 > control) that was partly attributed to the increased secretion of pro‐angiogenic factors VEGF and PDGF‐B. This is further supported by the evidence that pre‐treatment with inhibitor of VEGF receptor‐2 (Ki8751) or PDGF receptor (AG1296) throughout or only during the first 2‐day aortic ring culture period suppressed microvessel growth in GCCM‐treated groups, and also inhibited the FSH + TGFβ1‐GCCM‐stimulated release of matrix remodeling‐associated gelatinase activities. Interestingly, pre‐treatment of AG1296 at late stage suppressed GCCM‐induced microvessel growth and stability with demise of endothelial and mural cells. Together, we provide original findings that both FSH and TGFβ1 increased the secretion of VEGF and PDGF‐B, and that in turn up‐regulated the angiogenic activity in rat ovarian granulosa cells. This implicates that FSH and TGFβ1 play important roles in regulation of ovarian angiogenesis during follicle development. J. Cell. Physiol. 226: 1608–1619, 2011. © 2010 Wiley‐Liss, Inc.
Involvement of membrane type 1‐matrix metalloproteinase (MT1‐MMP) in RAGE activation signaling pathways
Authors: Kamioka Masashi , Ishibashi Toshiyuki , Ohkawara Hiroshi , Nagai Ryoji , Sugimoto Koichi , Uekita Hironori , Matsui Takanori , Yamagishi Sho‐Ichi , Ando Katsuya , Sakamoto Takayuki , Sakamoto Nobuo , Takuwa Yoh , Wada Ikuo , Shiomi Masashi , Maruyama Yukio , Takeishi Yasuchika
Published: October 2010

An advanced glycation end products (AGE)/a receptor for AGE (RAGE) axis plays a key role in diabetic vascular complications. Membrane type 1‐matrix metalloproteinase (MT1‐MMP) has been shown to function not only as a proteolytic enzyme but also as a signaling molecule. In this study, we investigated the role of MT1‐MMP in the AGE/RAGE‐triggered signaling pathways in cultured rabbit smooth muscle cells (SMCs) and the molecular interaction between RAGE and MT1‐MMP in vitro and in vivo. In SMCs, AGE‐activated Rac1 and p47phox within 1 min, NADPH oxidase activity and reactive oxygen species (ROS) generation within 5 min, and NF‐κB phosphorylation within 15 min, thereby inducing redox‐sensitive molecular expression. Silencing of RAGE by small‐interfering RNA (siRNA) blocked the AGE‐induced signaling pathways. AGE‐induced geranylgeranyl transferase I (GGTase I) activity, Rac1·p47phox activation, NADPH oxidase activity, ROS generation, and molecular expression were also markedly attenuated by silencing of MT1‐MMP. An inhibitor of GGTase I mimicked the effects of MT1‐MMP‐specific siRNA. Fluorescent immunohistochemistry revealed that MT1‐MMP was partially co‐localized with RAGE in SMCs, and RAGE was found to form a complex with MT1‐MMP in both cultured SMCs and the aortae of diabetic rats by immunoprecipitation. Furthermore, MT1‐MMP and RAGE formed a complex in the aortic atherosclerotic lesions of hyperlipidemic rabbits. We show that MT1‐MMP plays a crucial role in RAGE‐activated NADPH oxidase‐dependent signaling pathways and forms a complex with RAGE in the vasculature, thus suggesting that MT1‐MMP may be a novel therapeutic target for diabetic vascular complications. J. Cell. Physiol. 226: 1554–1563, 2011. © 2010 Wiley‐Liss, Inc.
Accuracy of bedside physical examination in distinguishing categories of shock
Authors: Vazquez Rodrigo , Gheorghe Cristina , Kaufman David , Manthous Constantine A.
Published: October 2010

HYPOTHESIS:

Selected elements of a simple physical examination algorithm accurately predict categories of shock.

SETTING:

A 350‐bed community teaching hospital.

METHODS:

Resident trainees who manage all critically ill and medically unstable patients were instructed to document capillary refill, (palpated) pulse volume, skin temperature, jugular venous pressure (JVP) and lung examination in all patients with prolonged (>30 minutes hypotension <90 mmHg). Treatment was determined by house officers guided by attending physicians of record. All cases were retrospectively reviewed by a senior clinician who applied consensus criteria/definitions to categorize shock as septic, cardiogenic or hypovolemic. Operating characteristics of examination findings for predicting categories of shock were computed.

RESULTS:

A total of 68 patients, averaging 71 ± 16 years, were studied. A total of 37 patients were diagnosed with septic shock, 18 with cardiogenic shock of and 13 with hypovolemic shock. Capillary refill and skin temperature predicted septic shock with sensitivity of 89%, specificity of 68%, positive predictive value (PPV) of 77%, negative predictive value (NPV) of 84%, and overall accuracy of 79%. Presence of JVP >7 cmH2O was more accurate than bilateral pulmonary crackles (>1/3 from bases) in predicting cardiogenic shock for low‐output patients with sensitivity of 82%, specificity of 79%, PPV of 75%, NPV of 85%, and overall accuracy of 80%. Using just skin temperature and JVP, the bedside approach correctly diagnosed 52/68 cases (overall accuracy = 76%).

CONCLUSIONS:

Simple bedside clinical examination findings correctly predict categories of shock in a majority of cases. Journal of Hospital Medicine 2010. © 2010 Society of Hospital Medicine.
Unplanned transfers to the intensive care unit: The role of the shock index
Authors: Keller A. Scott , Kirkland Lisa L. , Rajasekaran Smita Y. , Cha Stephen , Rady Mohamed Y. , Huddleston Jeanne M.
Published: October 2010

BACKGROUND:

Unplanned (unexpected) transfers to the intensive care unit (ICU) are typically preceded by physiologic instability. However, trends toward instability may be subtle and not accurately reflected by changes in vital signs. The shock index (SI) (heart rate/systolic blood pressure as an indicator of left ventricular function, reference value of 0.54) may be a simple alternative means to predict clinical deterioration.

OBJECTIVE:

To assess the association of the SI with unplanned ICU transfers.

DESIGN:

Retrospective case‐control study.

SETTING:

Academic medical center.

PATIENTS:

Fifty consecutive general medical patients with unplanned ICU transfers between 2003 and 2004 and 50 matched controls admitted to the same general medical unit between 2002 and 2004.

MEASUREMENTS:

Demographic data and vital signs abstracted from chart review.

RESULTS:

The SI was associated with unplanned ICU transfer at values of 0.85 or greater (P < 0.02; odds ratio, 3.0) and there was a significant difference between the median of worst shock indices of cases and controls (0.87 vs. 0.72; P< 0.005). There was no significant difference in age, race, admission ward, or Charlson Comorbidity Index, but hospital stay for cases was significantly longer (mean [standard deviation, SD], 14.8 [9.7] days vs. 5.7 [6.3] days; P < 0.001).

CONCLUSIONS:

SI is associated with unplanned transfers to the ICU from general medical units at values of 0.85 or greater. Future studies will determine whether SI is more accurate than simple vital signs as an indicator of clinical decline. If so, it may be a useful trigger to activate medical emergency or rapid response teams (RRTs). Journal of Hospital Medicine 2010. © 2010 Society of Hospital Medicine.
Redefining readmission risk factors for general medicine patients
Authors: Allaudeen Nazima , Vidyarthi Arpana , Maselli Judith , Auerbach Andrew
Published: October 2010

BACKGROUND:

Readmissions are costly both financially for our healthcare system and emotionally for our patients. Identifying factors that increase risk for readmissions may be helpful to focus resources to optimize the discharge process and reduce avoidable readmissions.

OBJECTIVE:

To identify factors associated with readmission within 30 days for general medicine patients.

METHODS:

We performed a retrospective observational study of an administrative database at an urban 550‐bed tertiary care academic medical center. Cohort patients were discharged from the general medicine service over a 2‐year period from June 1, 2006, to May 31, 2008. Clinical, operational, and sociodemographic factors were evaluated for association with readmission.

RESULTS:

Our cohort included 10,359 consecutive admissions (6805 patients) discharged from the general medicine service. The 30‐day readmission rate was 17.0%. In multivariate analysis, factors associated with readmission included black race (odds ratio [OR], 1.43; 95% confidence interval [CI], 1.24–1.65), inpatient use of narcotics (1.33; 1.16–1.53) and corticosteroids (1.24; 1.09–1.42), and the disease states of cancer (with metastasis 1.61; 1.33–1.95; without metastasis 1.95; 1.54–2.47), renal failure (1.19; 1.05–1.36), congestive heart failure (1.30; 1.09–1.56), and weight loss (1.26; 1.09–1.47). Medicaid payer status (1.15; 0.97–1.36) had a trend toward readmission.

CONCLUSION:

Readmission of general medicine patients within 30 days is common and associated with several easily identifiable clinical and nonclinical factors. Identification of these risk factors can allow providers to target interventions to reduce potentially avoidable readmissions. Journal of Hospital Medicine 2010. © 2010 Society of Hospital Medicine
Modification of Polydivinylbenzene Microspheres by a Hydrobromination/Click‐Chemistry Protocol and their Protein‐Adsorption Properties
Authors: Durmaz Yasemin Yuksel , Karagoz Bunyamin , Bicak Niyazi , Demirkol Dilek Odaci , Yalcinkaya Esra Evrim , Timur Suna , Yagci Yusuf
Published: October 2010

Hydrophobic‐ and/or hydrophilic‐polymer‐grafted PDVB microspheres are synthesized by the combination of hydrobromination and click‐chemistry processes. The modified‐PDVB microspheres and the intermediates at various stages of synthesis are characterized using GPC, 1H NMR and FTIR spectroscopy and TGA analysis. Use of the microspheres as a support matrix for reversible protein immobilization via adsorption is investigated. The system parameters such as the adsorption conditions (i.e., enzyme concentration, medium pH) and desorption are studied and evaluated with regards to the biocatalytic activity and adsorption capacity.

magnified image
Fabrication of a Thermoresponsive Biohybrid Double Hydrophilic Block Copolymer by a Cofactor Reconstitution Approach
Authors: Wan Xuejuan , Liu Shiyong
Published: October 2010

The fabrication of a thermoresponsive biohybrid double hydrophilic block copolymer (DHBC) by a cofactor reconstitution approach is reported. Poly(N‐isopropylacrylamide) (PNIPAM) bearing a porphyrin moiety at the chain terminal, PPIXZn‐PNIPAM, is synthesized by the combination of ATRP and a click reaction. The subsequent cofactor reconstitution process between apomyoglobin and PPIXZn‐PNIPAM affords well‐defined myoglobin‐b‐PNIPAM protein–polymer bioconjugates. Behaving as typical responsive DHBCs, the obtained myoglobin‐b‐PNIPAM biohybrid diblock copolymer exhibits thermo‐induced aggregation behavior in aqueous solution as a result of the presence of the thermoresponsive PNIPAM block, as revealed by temperature‐dependent transmittance, dynamic laser light scattering measurements, transmission electron microscopy, and scanning electron microscopy. This work represents the first report of the preparation of responsive biohybrid DHBCs by the cofactor reconstitution process.

magnified image
Heteronuclear chemical shift correlation and J‐resolved MAS NMR spectroscopy of lipid membranes
Authors: Zorin Vadim , Ciesielski Filip , Griffin David C. , Rittig Michael , Bonev Boyan B.
Published: October 2010

Direct observation of J‐couplings remains a challenge in high‐resolution solid‐state NMR. In some cases, it is possible to use Lee–Goldburg (LG) homonuclear decoupling during rare spin observation in MAS NMR correlation spectroscopy of lipid membranes to obtain J‐resolved spectra in the direct dimension. In one simple implementation, a wide line separation‐type 13C‐1H HETCOR can provide high‐resolution 1H/13C spectra, which are J‐resolved in both dimensions. Coupling constants, 1JHC, obtained from 1H doublets, can be compared with scaled 1JθCH‐values obtained from the 13C multiplets to assess the LG efficiency and scaling factor. The use of homonuclear decoupling during proton evolution, LG‐HETCOR‐LG, can provide J‐values, at least in the rare spin dimension, and allows measurements in less mobile membrane environments. The LG‐decoupled spectroscopic approach is demonstrated on pure dioleoylphosphatidylcholine (DOPC) membranes and used to investigate lipid mixtures of DOPC/cholesterol and DOPC/cholesterol/sphingomyelin. Copyright © 2010 John Wiley & Sons, Ltd.
Iopamidol as a responsive MRI‐chemical exchange saturation transfer contrast agent for pH mapping of kidneys: In vivo studies in mice at 7 T
Authors: Longo Dario Livio , Dastrù Walter , Digilio Giuseppe , Keupp Jochen , Langereis Sander , Lanzardo Stefania , Prestigio Simone , Steinbach Oliver , Terreno Enzo , Uggeri Fulvio , Aime Silvio
Published: October 2010

Iopamidol (Isovue®—Bracco Diagnostic Inc.) is a clinically approved X‐Ray contrast agent used in the last 30 years for a wide variety of diagnostic applications with a very good clinical acceptance. Iopamidol contains two types of amide functionalities that can be exploited for the generation of chemical exchange saturation transfer effect. The exchange rate of the two amide proton pools is markedly pH‐dependent. Thus, a ratiometric method for pH assessment has been set‐up based on the comparison of the saturation transfer effects induced by selective irradiation of the two resonances. This ratiometric approach allows to rule out the concentration effect of the contrast agent and provides accurate pH measurements in the 5.5–7.4 range. Upon injection of Iopamidol into healthy mice, it has been possible to acquire pH maps of kidney regions. Furthermore, it has been also shown that the proposed method is able to report about pH‐changes induced in control mice fed with acidified or basified water for a period of a week before image acquisition. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.
Hormone Changes Affecting Energy Homeostasis after Metabolic Surgery
Authors: Harvey E. John , Arroyo Kervin , Korner Judith , Inabnet William B.
Published: October 2010

Worldwide, there is an epidemic of obesity and its associated diseases. The past decade of research has brought about a paradigm shift in our understanding of both the mechanisms underlying energy homeostasis and the multiple factors contributing to the pathophysiology of obesity. Metabolic surgery is currently far more effective than diet and exercise or pharmacotherapy in achieving durable weight loss. Moreover, the remarkable results of surgery in achieving a rapid remission of type 2 diabetes mellitus has sparked tremendous excitement and research into the mechanisms through which metabolic surgery has its dramatic effect. As opposed to the traditional understanding of “restriction” and “malabsorption,” current evidence suggests that metabolic surgery alters the expression of multiple hormones that affect both short‐term and long‐term signals of energy balance. We review the hormonal changes following the most common types of metabolic operations currently being performed. The profile of hormonal changes provides a guide to tailor the choice of operation for each individual patient toward achieving the desired metabolic result. In the future, individualized metabolic surgery alone or modulated by targeted pharmacological therapy may achieve the most reliable and effective results with the highest safety and lowest side effect profile. Mt Sinai J Med 77:446–465, © 2010 Mount Sinai School of Medicine
Plasma proteome analysis in diet‐induced obesity‐prone and obesity‐resistant rats
Authors: Choi Jung‐Won , Wang Xia , Joo Jeong In , Kim Dong Hyun , Oh Tae Seok , Choi Duk Kwon , Yun Jong Won
Published: October 2010

One of the major issues in the field of obesity is why some humans become obese and others resist development of obesity when exposed to high‐calorie diets. Despite the same genetic background, namely obesity‐prone (OP) and ‐resistant (OR) rats, differing responses have been demonstrated in a high fat diet‐induced rodent model. The aim of the present study was to discover novel obesity‐related biomarkers for susceptibility and/or resistance to obesity by proteomic analysis of OP and OR rat plasma. After feeding of high fat diet, OP rats gained approximately 25% more body weight than OR rats and were used for proteomic analysis using 2‐DE combined with MALDI‐TOF‐MS. We categorized identified proteins into three groups by analysis of both average spot density in each group and individual spot density of six rats as a function of body weight. Consequently, category (1) included inter‐α‐inhibitor H4 heavy chain and fetuin B precursor, which can be used as novel plasma biomarkers for risk of obesity. Nine proteins of category (2) and (3) can also be plausible plasma markers in the study of obesity. This proteomic study is an important advancement over the previous steps needed for identification of OP and OR rats.
Effect of the Rho kinase inhibitor Y‐27632 on the proteome of hearts with ischemia–reperfusion injury
Authors: Cadete Virgilio J. J. , Sawicka Jolanta , Polewicz Dorota , Doroszko Adrian , Wozniak Mieczyslaw , Sawicki Grzegorz
Published: October 2010

Growing attention has been given to the role of the Rho kinase pathway in the development of heart disease and ischemia/reperfusion (I/R) injury. Y‐27632 is a Rho kinase inhibitor demonstrated to protect against I/R injury, but the exact mechanism by which it does so remains to be elucidated. The goal of this project was to determine new targets by which Y‐27632 can protect the heart against I/R injury. Isolated rat hearts were perfused under aerobic conditions or subjected to I/R in the presence or absence of Y‐27632. Administration of Y‐27632 (1 μM) before ischemia and during the first 10 min of reperfusion resulted in complete recovery of cardiac function. 2‐D electrophoresis followed by MS identified four proteins whose levels were affected by Y‐27632 treatment. Lactate dehydrogenase and glyceraldehyde‐3‐phosphate dehydrogenase were significantly increased in the Y‐27632 treated group, while creatine kinase was normalized to control levels. In addition, we found increased level of two different molecular fragments of ATP synthase, which were normalized by Y‐27632. This increase suggests that during ischemia ATP synthase is subjected to degradation. The changes in metabolic enzymes' levels and their regulation by Y‐27632 suggest that the cardioprotective effect of Y‐27632 involves increased energy production.
Mass spectrometry characterization of 3‐OH butyrated β‐cyclodextrin
Authors: Peptu Cristian , Nicolescu Alina , Peptu Catalina A. , Harabagiu Valeria , Simionescu Bogdan C. , Kowalczuk Marek
Published: October 2010

Oligo(3‐OH butyrate)‐β‐cyclodextrin esters (PHB‐CD) were obtained through ring opening of β‐butyrolactone (β‐BL) in the presence of β‐cyclodextrin (CD) and (‐)‐sparteine (SP) as nucleophilic activator. The resulted reaction mixture was first separated in two fractions and then investigated through a deep mass spectrometry (MS) study performed on a liquid chromatography‐electrospray ionization‐quadrupole time of flight (LC‐ESI‐QTOF) instrument. LC MS and tandem MS structural assignment of the reaction products was completed by NMR. The performed analysis revealed that poly(3‐OH butyrate) homopolymers (PHB) are formed together with the PHB‐CD products. Secondary reactions resulting in the formation of crotonates were also proved to occur. A comparison between MS and NMR results demonstrated that more than one PHB oligomer is attached to the CD in the PHB‐CD product. The tandem MS fragmentation studies validated the proposed structure of CD derivatives. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2010
Rapid exploration of the folding topology of helical membrane proteins using paramagnetic perturbation
Authors: Yeo Kwon Joo , Kim Hye‐Yeon , Kim Young Pil , Hwang Eunha , Kim Myung Hee , Cheong Chaejoon , Choe Senyon , Jeon Young Ho
Published: October 2010

An understanding of the folding states of α‐helical membrane proteins in detergent systems is important for functional and structural studies of these proteins. Here, we present a rapid and simple method for identification of the folding topology and assembly of transmembrane helices using paramagnetic perturbation in nuclear magnetic resonance spectroscopy. By monitoring the perturbation of signals from glycine residues located at specific sites, the folding topology and the assembly of transmembrane helices of membrane proteins were easily identified without time‐consuming backbone assignment. This method is validated with Mistic (membrane‐integrating sequence for translation of integral membrane protein constructs) of known structure as a reference protein. The folding topologies of two bacterial histidine kinase membrane proteins (SCO3062 and YbdK) were investigated by this method in dodecyl phosphocholine (DPC) micelles. Combing with analytical ultracentrifugation, we identified that the transmembrane domain of YbdK is present as a parallel dimer in DPC micelle. In contrast, the interaction of transmembrane domain of SCO3062 is not maintained in DPC micelle due to disruption of native structure of the periplasmic domain by DPC micelle.