bluuebunny/crossref_metadata_2025 huggingface dataset:
ADAMTS‐1–Knockout mice do not exhibit abnormalities in aggrecan turnover in vitro or in vivo
Authors: Little Chris B. , Mittaz Laureane , Belluoccio Daniele , Rogerson Fraser M. , Campbell Ian K. , Meeker Clare T. , Bateman John F. , Pritchard Melanie A. , Fosang Amanda J.
Published: May 2005

Objective

To determine the role of the proteinase ADAMTS‐1 in normal and accelerated catabolism of aggrecan in articular and growth plate cartilage of mice.

Methods

Expression of ADAMTS‐1 was determined using reverse transcriptase–polymerase chain reaction (RT‐PCR) analysis of RNA isolated from microdissected chondrocytes from different zones of mouse growth plate and articular cartilage. Real‐time RT‐PCR for ADAMTS‐4, ADAMTS‐5, and ADAMTS‐9 was performed on femoral head cartilage of wild‐type (WT) and ADAMTS‐1–knockout (KO) mice. Histologic and immunohistologic evaluation of growth plate and articular cartilage was performed in WT and KO mice from birth to 12 weeks of age. The effect of ADAMTS‐1 ablation on cartilage proteoglycan loss was studied in antigen‐induced arthritis (AIA). Aggrecan catabolism in WT and KO mice was studied in an in vitro model of cartilage degradation, by quantitation of glycosaminoglycan loss and histologic, immunohistologic, and Western immunoblot analyses.

Results

ADAMTS‐1 messenger RNA (mRNA) was expressed in normal mouse articular and growth plate cartilage and was up‐regulated in terminal hypertrophic differentiation of growth plate chondrocytes. There was no difference in mRNA levels in the cartilage of WT compared with KO mice for the other potential aggrecanases ADAMTS‐4, ADAMTS‐5, or ADAMTS‐9. ADAMTS‐1–KO mice were significantly smaller than their WT littermates; however, no morphologic differences between the genotypes were evident in growth plate or articular cartilage from birth to skeletal maturity (12–16 weeks). Similarly, no difference in cartilage aggrecan content or presence of aggrecan degradation products was detected between WT and KO mice. There was no difference between WT and KO mice in the degree of synovial inflammation or depletion of cartilage aggrecan in AIA. There was no difference between WT and KO cartilage in either basal or stimulated aggrecan loss in vitro; however, subtle changes in the aggrecanase‐generated aggrecan catabolites were observed in interleukin‐1–treated cartilage.

Conclusion

Although ADAMTS‐1 is expressed in articular and growth plate cartilage and is able to cleave aggrecan at physiologically relevant sites, our results indicate that it does not play a significant nonredundant role in normal cartilage and bone development and growth. Similarly, ablation of ADAMTS‐1 offered no protection from accelerated aggrecanolysis in an inflammatory model of arthritis or in an in vitro model of early cartilage degradation. ADAMTS‐1 does not appear to be a viable target for treatment of cartilage destruction in arthritis.
Selection of knee radiographs for trials of structure‐modifying drugs in patients with knee osteoarthritis: A prospective, longitudinal study of lyon schuss knee radiographs with the definition of adequate alignment of the medial tibial plateau
Authors: Conrozier Thierry , Mathieu Pierre , Piperno Muriel , Favret Huguette , Colson Frédéric , Vignon Martine , Conrozier Sylvie , Vignon Eric
Published: May 2005

Objective

The quality of medial tibial plateau (MTP) alignment, which is assessed by measuring the distance between the anterior and posterior margins (intermargin distance [IMD]) of the tibial plateau, and the reproducibility of alignment in serial radiographs are suggested to be key elements in determining the accuracy and sensitivity to change in knee radiographs in patients with tibiofemoral osteoarthritis (OA). We evaluated the influence of both MTP alignment and radiograph superimposition on the sensitivity to change in radiographic joint space narrowing (JSN) in knee OA.

Methods

The study group comprised 106 patients with knee pain (73 with OA). Lyon schuss radiographic images of the knee were obtained twice (at baseline [month 0] and 12 months later), using a standardized radiographic procedure. Computerized measurement of the IMD for the assessment of MTP alignment was compared with the grading of MTP alignment by 2 observers using a 5‐point scale (excellent, good, fair, poor, bad). To obtain the rate of JSN, computerized measurement of the joint space width was performed at month 0 and month 12. The sensitivity of the joint space width to change over 1 year was evaluated by the standardized response mean (SRM).

Results

The mean (±SD) IMD was 1.2 ± 0.9 mm. The correlation between scoring and computer measurement of MTP alignment was highly significant. The cutoff value for satisfactory alignment (excellent or good) was an IMD of ≤1.2 mm. In OA knees, the mean (±SD) annual rate of JSN and the SRM were statistically higher in knees with an IMD of ≤1.2 mm at both month 0 and month 12 (0.34 ± 0.50 mm and 0.68, respectively) than in knees with an IMD of >1.2 mm at month 0 and/or month 12.

Conclusion

The quality of MTP alignment at both baseline and the end point highly influences the sensitivity to change in radiographic JSN in knee OA. To obtain relevant data, only radiographs showing an IMD of ≤1.2 mm at both baseline and the end point would have to be analyzed in studies of structure‐modifying OA drugs.
Activation of the interferon‐α pathway identifies a subgroup of systemic lupus erythematosus patients with distinct serologic features and active disease
Authors: Kirou Kyriakos A. , Lee Christina , George Sandhya , Louca Kyriakos , Peterson Margaret G. E. , Crow Mary K.
Published: May 2005

Objective

Gene‐expression studies have demonstrated increased expression of interferon (IFN)–inducible genes (IFIGs) in peripheral blood mononuclear cells (PBMCs) of many patients with systemic lupus erythematosus (SLE), with a predominant effect of type I IFN. This study examined the hypothesis that increased disease severity and activity, as well as distinct autoantibody specificities, characterize SLE patients with activation of the type I IFN pathway.

Methods

Freshly isolated PBMCs from 77 SLE patients, 22 disease controls, and 28 healthy donors were subjected to real‐time polymerase chain reaction for 3 IFIGs that are preferentially induced by IFNα, and the data were used to derive IFNα scores for all individuals. Expression of IFIGs was significantly higher in SLE patients compared with disease controls or healthy donors. SLE patients with high and low IFNα scores were compared for clinical manifestations of disease, disease severity, disease activity, serologic features, and potential confounders, by bivariate and multivariate analyses.

Results

SLE patients with a high IFNα score had a significantly higher prevalence of renal disease, a greater number of American College of Rheumatology criteria for SLE, and a higher Systemic Lupus International Collaborating Clinics damage index (SDI) score than did SLE patients with low IFNα scores. Patients with high scores showed increased disease activity, as measured by lower C3 levels, hemoglobin levels, absolute lymphocyte counts, and albumin levels, and a higher anti–double‐stranded DNA (dsDNA) titer, erythrocyte sedimentation rate, and SLE Disease Activity Index 2000 score. The presence of antibodies specific for Ro, U1 RNP, Sm, and dsDNA, but not phospholipids, was significantly associated with a high IFNα score. Logistic regression analysis confirmed that renal disease, higher SDI scores, low complement levels, and presence of anti–RNA binding protein (RBP) autoantibodies were associated with a high IFNα score.

Conclusion

Activation of the IFNα pathway defines a subgroup of SLE patients whose condition is characterized by increased disease severity, including renal disease, increased disease activity, reflected in complement activation, and autoreactivity to RBP.
T cell costimulation by fractalkine‐expressing synoviocytes in rheumatoid arthritis
Authors: Sawai Hirokazu , Park Yong Wook , Roberson James , Imai Toshio , Goronzy Jörg J. , Weyand Cornelia M.
Published: May 2005

Objective

Patients with rheumatoid arthritis (RA) accumulate prematurely aged T cells that have acquired a new profile of regulatory receptors. Many of the de novo–expressed receptors are typically found on natural killer cells, including CX3CR1, the receptor for the chemokine fractalkine (FKN). This study explored whether interactions between CX3CR1 and FKN are relevant for T cell functions in rheumatoid synovitis.

Methods

FKN expression was examined by real‐time polymerase chain reaction and immunohistochemistry. CX3CR1 expression on peripheral blood T cells was analyzed by flow cytometry. T cell activation was quantified by determining proliferative responses, interferon‐γ (IFNγ) secretion, and granule release. Fibroblast‐like synoviocyte (FLS)/T cell adhesion was measured by the retention of 5‐carboxyfluorescein diacetate succinimidyl ester–labeled T cells on FLS monolayers.

Results

FKN was expressed on cultured synovial fibroblasts and hyperplastic synoviocytes in the rheumatoid tissue. Among CD4+ T cells, only senescent CD28− T cells were positive for CX3CR1 (P < 0.001). Such CD4+,CD28−,CX3CR1+ T cells strongly adhered to FLS, with soluble FKN blocking the interaction. FKN expressed on FLS costimulated T cell–activating signals and amplified proliferation, IFNγ production, and expulsion of cytoplasmic granules.

Conclusion

Senescent CD4+ T cells that accumulate in rheumatoid arthritis aberrantly express CX3CR1. FKN, which is membrane‐anchored on synoviocytes, enhances CD4+ T cell adhesion, provides survival signals, and costimulates the production of proinflammatory cytokines as well as the release of granules. By virtue of their altered receptor profile, senescent CD4+ T cells receive strong stimulatory signals from nonprofessional antigen‐presenting cells in the synovial microenvironment.
Factor matrix text filtering and clustering
Authors: Kostoff Ronald N. , Block Joel A.
Published: May 2005

The presence of trivial words in text databases can affect record or concept (words/phrases) clustering adversely. Additionally, the determination of whether a word/phrase is trivial is context‐dependent. Our objective in the present article is to demonstrate a context‐dependent trivial word filter to improve clustering quality. Factor analysis was used as a context‐dependent trivial word filter for subsequent term clustering. Medline records for Raynaud's Phenomenon were used as the database, and words were extracted from the record abstracts. A factor matrix of these words was generated, and the words that had low factor loadings across all factors were identified, and eliminated. The remaining words, which had high factor loading values for at least one factor and therefore were influential in determining the theme of that factor, were input to the clustering algorithm. Both quantitative and qualitative analyses were used to show that factor matrix filtering leads to higher quality clusters and subsequent taxonomies.
An intervention study to increase knowledge and use of folic acid among relatives in neural tube defect–affected families in Washington, D.C.
Authors: Byrne Julianne , Carolan Susan , Arcement Rashad , Kozlowski Matthew , Taller Inna , Ried Stephanie , Keating Robert
Published: May 2005

BACKGROUND

Little is known about the level of knowledge and use of folic acid among near relatives in U.S. families of a child with spina bifida. We hypothesized that relatives would be more knowledgeable than the general population and more likely to take folic acid. Further, we hypothesized that relatives would be more motivated by an intervention to increase their use of folic acid.

METHODS

We conducted an intervention study among females in families attending a hospital spina bifida clinic in Washington, DC.

RESULTS

The 231 subjects consisted of the affected individuals, mothers, sisters, and aunts. The average age was 34 years. At baseline, most (87.4%) reported that they had heard of folic acid; 37.6% were currently taking multivitamins with folic acid and 6.9% were taking folic acid tablets. The intervention significantly increased both knowledge (to 99%) and intake of folic acid from 41.9 to 48.5%. Folic acid intake increased significantly among African‐American women and women with less education, women who were older, married, with children, and nonsmokers.

CONCLUSIONS

This intervention was successful in increasing folic acid intake among female relatives in spina bifida–affected families. By the end of the study, almost all women had heard of folic acid and folic acid use had increased by 16%. Among these women at higher than expected risk for having an affected child, this rate of intake, while more than the general population, still falls short of optimum. Fortification of food with folic acid may be the only way to ensure increased folic acid intake. Birth Defects Research (Part A), 2005. © 2005 Wiley‐Liss, Inc.
Methamphetamine and lipid peroxidation in the rat retina
Authors: Melo Pedro , Rodrigues Lorena G. , Pinazo‐Durán Maria Dolores , Tavares Maria Amélia
Published: May 2005

BACKGROUND

The use of psychoactive drugs during adolescence and early adult life has increased in the last few decades. It is known that developmental exposure to psychostimulants affects the sensory systems, and the retina has been shown to be a target tissue. This work was conducted to evaluate the pattern of lipid peroxidation in the rat retina following prenatal exposure to methamphetamine (MA).

METHODS

Pregnant female Wistar rats were given MA (5 mg/kg of body weight/day; SC, in 0.9% saline) from GD 8 to 22. Offspring were sacrificed at postnatal days (PNDs) 7, 14, and 21. The retinas were homogenized, and both the total antioxidant and superoxide dismutase (SOD) activities were measured by enzymatic‐colorimetric methods. The lipid peroxidation byproducts (malondialdehyde [MDA] and MDA‐like metabolites) were measured by the thiobarbituric acid test.

RESULTS

Total antioxidant levels were lower in the MA group at PND 21 in both males and females. The activity of SOD was higher in PND 7 females from the MA group. MDA levels were higher in the MA group at PND 21 in both genders.

CONCLUSIONS

These findings suggest that prenatal‐induced MA toxicity in the retina may be related to lipid peroxidation processes and oxidative stress. Birth Defects Research (Part A), 2005. © 2005 Wiley‐Liss, Inc.
Synthesis and conformational properties of protein fragments based on the Id family of DNA‐binding and cell‐differentiation inhibitors
Authors: Kiewitz Sebastian D. , Cabrele Chiara
Published: May 2005

Id proteins are dominant negative regulators of the helix–loop–helix (HLH) transcription factors and are important during development, especially by preventing cell differentiation while inducing cell proliferation. In contrast, they are poorly expressed in healthy adults but are found in several tumor types. The Id HLH motif is responsible for the inhibitory activity, whereas not much is known about the role of the N‐ and C‐termini. In the presented work, synthetic peptides reproducing the HLH, the N‐terminal region, and the C‐terminal region of the Id proteins were characterized by CD. The four HLH sequences built highly stable helical conformations, whereas the N‐ and C‐termini were unstructured, with the exception of an alanine‐rich fragment preceding the Id4 HLH motif. Deletion of the loop connecting the two helices led to helix destabilization for all four Id HLH peptides. In addition, modifications of the amino acid composition within the hydrophobic face of the helices of the Id1 HLH peptide induced conformational changes, mostly associated with loss of helix content. Moreover, a fragment containing the helix‐2 and the C‐terminus of the Id1 protein did not show any helical character. Therefore, both the helix propensity and stability of the HLH domain were shown to be strongly dependent on favorable interhelical contacts. In contrast, it is suggested that the regions beyond this domain could rather play a destabilizing role, for example, by increasing the flexibility of the folded protein. © 2005 Wiley Periodicals, Inc. Biopoly 80: 762–774, 2005

This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
Efficient Calculation of Many Stacking and Pairing Free Energies in DNA from a Few Molecular Dynamics Simulations
Authors: Oostenbrink Chris , van Gunsteren Wilfred F.
Published: May 2005

Through the use of the one‐step perturbation approach, 130 free energies of base stacking and 1024 free energies of base pairing in DNA have been calculated from only five simulations of a nonphysical reference state. From analysis of a diverse set of 23 natural and unnatural bases, it appears that stacking free energies and stacking conformations play an important role in pairing of DNA nucleotides. On the one hand, favourable pairing free energies were found for bases that do not have the possibility to form canonical hydrogen bonds, while on the other hand, good hydrogen‐bonding possibilities do not guarantee a favourable pairing free energy if the stacking of the bases dictates an unfavourable conformation. In this application, the one‐step perturbation approach yields a wealth of both energetic and structural information at minimal computational cost.
Synthesis of Multi‐1,10‐phenanthroline Ligands with 1,3‐Phenylene Linkers and Their Lithium Complexes
Authors: Dietrich‐Buchecker Christiane , Colasson Benoît , Jouvenot Damien , Sauvage Jean‐Pierre
Published: May 2005

The synthesis of two multisite ligands containing four and five 1,10‐phenanthroline (phen) chelates in line, respectively, is presented. The connectors are 1,3‐phenylene linkers. The two ligands were prepared following multistep procedures, the two key reactions being the Suzuki coupling reaction between aromatic nuclei and the nucleophilic addition of aryllithium derivatives onto a phen fragment. The coordination chemistry of both ligands with Li+ions was very clean and selective, whereas their reaction with copper(I) led to intractable mixtures of insoluble complexes. The tetraphen and the pentaphen compounds afforded almost quantitatively the four‐ and five‐lithium double‐stranded helical complexes, respectively. The helical systems are probably highly wound, as indicated by NMR measurements. The pronounced strain of the 5‐Li+complex is reflected by the easy loss of a lithium cation, as shown by electrospray mass spectrometry.
The HIV‐Tat protein induces chromosome number aberrations by affecting mitosis
Authors: Battaglia Piero A. , Ponti Donatella , Naim Valeria , Venanzi Serenella , Psaila Rossana , Gigliani Franca
Published: May 2005

To analyze the effects of the HIV‐Tat‐tubulin interaction, we microinjected HIV‐Tat purified protein into Drosophila syncytial embryos. Following the Tat injection, altered timing of the cortical nuclear cycles was observed; specifically, the period between the nuclear envelope breakdown and anaphase initiation was lengthened as was the period between anaphase initiation and the formation of the next nuclear envelope. These two periods correspond to kinetochore alignment at metaphase and to mitosis exit, respectively. We also demonstrated that these two delays are the consequence of damage specifically induced by Tat on kinetochore alignment and on the timing of sister chromatid segregation at anaphase. Furthermore, we show that the expression of Tat in Drosophila larvae brain cells produces a significant percentage of polyploid and aneuploid cells. The results reported here indicate that Tat impairs the mitotic process and that Tat‐tubulin interaction appears to be responsible for the observed defects. The presence of polyploid and aneuploid cells is consistent with a delay or arrest in the M phase of a substantial fraction of the cells expressing Tat, suggesting that mitotic spindle checkpoints are overridden following Tat expression. Cell Motil. Cytoskeleton 61:129–136, 2005. © 2005 Wiley‐Liss, Inc.
Synthesis of Poly(ε‐lysine)‐Grafted Dextrans and Their pH‐ and Thermosensitive Hydrogelation with Cyclodextrins
Authors: Choi Hak Soo , Yamamoto Kaori , Ooya Tooru , Yui Nobuhiko
Published: May 2005

To give pH sensitivity to a thermoreversible supramolecular‐structured hydrogel system, poly(ε‐lysine) (PL), as a cationic polymer, was grafted to dextran and used for inclusion complexation with α‐cyclodextrins (α‐CDs). The synthesized graft copolymer was characterized by 1H NMR spectroscopy, and the hydrogel formation was confirmed by X‐ray diffraction and solid‐state 13C NMR analysis. The hydrogelation was induced from a phase‐separated structure of hydrated dextrans and hydrophobically aggregated inclusion complexes in buffer solution at pH 10.0. The prepared hydrogels showed thermoreversible gel–sol transitions as well as pH‐sensitive phase transitions, which were recorded by the changes in UV/Vis transmittance. A rapid phase transition from gel to sol was observed upon decreasing the pH value to 4.0, which resulted from the dissociation process between the protonated guest polymer and α‐CDs. The stimuli‐responsive physical properties of the hydrogels were improved by modulating the degree of substitution of the grafted PL and the combination with α‐CDs.
The insulin sensitivity, glucose sensitivity, and acute insulin response to glucose load in adolescent type 2 diabetes in Taiwanese
Authors: Pei Dee , Hsiao Chin‐Fu , Hung Yi‐Jen , Hsieh Chang‐Hsun , Fang Sandra‐Chi , Lian Wei‐Chen , Hsu Wen‐Lin , Fu Chen‐Chung , Chen Hsin‐Dean , Kuo Shi‐Wen
Published: May 2005

Background

Insulin sensitivity (SI), glucose sensitivity (SG), acute insulin response to glucose load (AIR), and obesity in adolescent type 2 diabetes patients (young diabetes, YDM) in Taiwan were studied.

Methods

Forty patients diagnosed at <22 years of age were enrolled and divided into non‐obese (NOYDM, BMI < 27 kg/m2) and obese groups (OBYDM BMI > 27 kg/m2). Adult‐onset type 2 diabetes patients (ADM) >40 years old (n = 41) and nondiabetic young adults (N) (n = 23) served as controls. Fasting plasma lipids, insulin, and glucose were measured. Homeostasis model assessment was calculated to estimate insulin sensitivity and β‐cell function. A frequent‐sampled intravenous glucose tolerance test was performed to evaluate SI, SG, and AIR.

Results

SI and AIR were significantly lower in YDM and ADM than in N (0.92 ± 0.13, 0.8 ± 0.15 and 3.24 ± 0.47 × 10−4/U/mL for SI; 40.3 ± 20.3, 107.3 ± 50.2, 1208 ± 306.3 uU/min for AIR). SG of YDM and ADM were lower compared with N (0.014 ± 0.00138, 0.0292 ± 0.0058 vs 0.034 ± 0.0086 min−1 respectively). No difference was noted between YDM and ADM. SI and SG were not different in NOYDM and OBYDM. AIR was higher in OBYDM (83.6 ± 34.3 vs −7.6 ± 13.66 uU/min).

Conclusions

YDM had defects in SI, SG, and AIR compared to N, which was similar to the pathophysiology of ADM. The results imply that YDM may be either a different subtype of diabetes or the same type of diabetes as ADM, with severe defects associated with earlier age of onset. OBYDM had higher AIR than NOYDM. Copyright © 2005 John Wiley & Sons, Ltd.
Cloning and expression analysis of pos‐1 in the nematodes Caenorhabditis briggsae and Caenorhabditis remanei
Authors: Konwerski Jamie , Senchuk Megan , Petty Emily , Lahaie David , Schisa Jennifer A.
Published: May 2005

The Caenorhabditis elegans pos‐1 gene encodes a zinc‐finger protein that is required for germline specification during embryogenesis. The maternally provided mRNA is translationally regulated both spatially and temporally during early development. We have cloned orthologs of pos‐1 from C. briggsae and C. remanei, two Caenorhabditis species that have diverged from C. elegans by approximately 20–40 million years. Two regions in the 3′ untranslated region are highly conserved among all three species. We find that the pos‐1 RNA is expressed in the hermaphrodite and female gonads of C. briggsae and C. remanei but POS‐1 protein is not detected at high levels in C. briggsae until the 2‐cell stage of embyrogenesis. The protein expression is restricted to the germline precursors of the embryo. We conclude that pos‐1 appears to be translationally regulated in C. briggsae as it is in C. elegans and speculate the conserved 3′ UTR sequences may be involved. Developmental Dynamics 233:1006–1012, 2005. © 2005 Wiley‐Liss, Inc.
A large Norwegian family with inherited malignant melanoma, multiple atypical nevi, and CDK4 mutation
Authors: Molven Anders , Grimstvedt Magne B. , Steine Solrun J. , Harland Mark , Avril Marie‐Françoise , Hayward Nicholas K. , Akslen Lars A.
Published: May 2005

Mutations in two loci encoding cell‐cycle‐regulatory proteins have been shown to cause familial malignant melanoma. About 20% of melanoma‐prone families bear a mutation in the CDKN2A locus, which encodes two unrelated proteins, p16INK4A and p14ARF. Mutations in the other locus, CDK4, are much rarer and have been linked to the disease in only three families worldwide. In the 1960s, a large Norwegian pedigree with multiple atypical nevi and malignant melanomas was identified. Subsequently, six generations and more than 100 family members were traced and 20 cases of melanoma verified. In this article, we report that CDK4 codon 24 is mutated from CGT to CAT (Arg24His) in this unusually large melanoma kindred. Intriguingly, one of the family members had ocular melanoma, but the CDK4 mutation could not be detected in archival tissue samples from this subject. Thus, the case of ocular melanoma in this family was sporadic, suggesting an etiology different from that of the skin tumors. The CDK4 mutation in the Norwegian family was identical to that in melanoma families in France, Australia, and England. Haplotype analysis using microsatellite markers flanking the CDK4 gene and single‐nucleotide polymorphisms within the gene did not support the possibility that there was a common founder, but rather indicated at least two independent mutational events. All CDK4 melanoma families known to date have a substitution of amino acid 24. In addition to resulting from selection pressure, this observation may be explained by the CG dinucleotide of codon 24 representing a mutational hot spot in the CDK4 gene. © 2005 Wiley‐Liss, Inc.
Ubiquitous expression of mRFP1 in transgenic mice
Authors: Zhu Huaxing , Wang Guangxue , Li Gang , Han Min , Xu Tian , Zhuang Yuan , Wu Xiaohui
Published: May 2005

Fluorescent proteins provide a powerful means to track gene expression and cellular behaviors in the study of model organisms such as mice. Among the new generation of fluorescent protein markers, the monomeric red fluorescent protein mRFP1 is particularly attractive because of its rapid maturation and minimal interference with GFP and GFP‐derived markers. Here we evaluate the utility of mRFP1 as a marker in transgenic mice. We show that high level and ubiquitous expression of mRFP1 does not affect mouse development, general physiology, or reproduction. mRFP1 expression can be readily detected with unaided eyes under daylight in transgenic mice on the albino background. The intensity of mRFP1 signals can be used to distinguish homozygous and heterozygous transgenic mice. Together, these features make mRFP1 an attractive marker for broad applications in transgenic research. genesis 42:86–90, 2005. © 2005 Wiley‐Liss, Inc.
Mechanism(s) of increased vascular cell adhesion on nanostructured poly(lactic‐co‐glycolic acid) films
Authors: Miller Derick C. , Haberstroh Karen M. , Webster Thomas J.
Published: May 2005

Studies have shown that poly(lactic‐co‐glycolic acid) (PLGA) films with nanometer surface features promote vascular endothelial and smooth muscle cell adhesion. The objective of this in vitro research was to begin to understand the mechanisms behind this observed increase in vascular cell adhesion. Results provided evidence that nanostructured PLGA adsorbed significantly more vitronectin and fibronectin from serum compared to conventional (or those not possessing nanometer surface features) PLGA. When separately preadsorbing both vitronectin and fibronectin, increased vascular smooth muscle and endothelial cell density was observed on nanostructured (compared to conventional) PLGA. Additionally, blocking of cell‐binding epitopes of fibronectin and vitronectin significantly decreased vascular cell adhesion on nanostructured (compared to conventional) PLGA. For this reason, results of the present in vitro study demonstrated that cell adhesive proteins adsorbed in different quantities and altered bioactivity on nanostructured compared to conventional PLGA topographies, which (at least in part) may account for the documented increased vascular cell adhesion on nanostructured PLGA. In this manner, this study continues to provide evidence for the promise of nanostructured PLGA in vascular tissue engineering applications. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
Apheresis activity in Venezuela
Authors: Saltiel Christiane
Published: May 2005

Interest for apheresis activity has been growing in Venezuela. In 1976 there were only a few devices; in 2003, 80 apheresis machines performed 27,675 donor apheresis procedures and 547 therapeutic procedures countrywide. We report the activity at the Metropolitan Blood Bank (the largest one of the country) in the period 1999–2003: 597 therapeutic procedures were performed in 171 patients, during 212 crisis episodes. The average age was 38 ± 16 years, 65% male and 35% female. Most of the therapeutic procedures were therapeutic plasma exchange for hematology diseases (mainly thrombotic thrombocitopenic purpura and hemophilia inhibitors), including 184 therapeutic procedures with the Autopheresis‐C (Baxter Healthcare Corp., Deerfield, IL). Most common adverse effects (3.9%) were hypotension and allergic reactions to the plasma. J. Clin. Apheresis. © 2005 Wiley‐Liss, Inc.
Apheresis in the Philippines
Authors: Narciso Carmen T.
Published: May 2005

This is a retrospective study of 131 patients who underwent 479 therapeutic apheresis (TA) procedures at St. Luke's Medical Center, in the Philippines, from 1994 to 2003. Diseases were classified according to indication categories using the American Society for Apheresis (ASFA) guidelines. Seventy‐eight percent of the patients belonged to ASFA Category I, 14% to Category II, 8% to Category III, and none in Category IV. The most common indication was for neurological disorders (62%), mainly acute inflammatory demyelinating polyneuropathy (49%) and myasthenia gravis (40%). The remaining disease groups were hematologic (25%), renal and metabolic (8%), and autoimmune and rheumatic diseases (5%). Oral corticosteroids are the preference of many for the treatment of CIDP, being much less expensive than IVIG infusion or TA. Favorable response is noted in the use of the latter to prevent kidney transplant rejection through antibody removal among pre‐sensitized patients before transplantation. Complete recovery was seen in 2 of 4 patients with acute hepatic failure. The negative outcome of controlled studies in the literature has limited our utilization of TA for the treatment of systemic lupus erythematosus. Immunoadsorption for idiopathic thrombocytopenic purpura has not been performed locally due to the unavailability and prohibitive cost of affinity columns. Factors affecting the practice of TA in our country are cited. Organization of a multicenter study group and an apheresis national registry may lead to a better future of TA in the Philippines. J. Clin. Apheresis © 2005 Wiley‐Liss, Inc.
Post‐transcriptional gene regulation by gamma herpesviruses
Authors: Swaminathan Sankar
Published: May 2005

The Epstein–Barr virus (EBV) SM protein is a member of a highly conserved family of proteins present in most mammalian herpes viruses. There is a significant amount of functional and sequence divergence among the homologs encoded by the human herpes viruses, including differences in mechanism of action and varying effects on splicing and transcription. Nevertheless, in those cases where it has been studied, these proteins are essential for lytic replication of the virus. The mechanism by which SM regulates gene expression operates at the level of mRNA stability, processing, and export. SM enhances expression of EBV lytic genes and has both positive and negative effects on cellular gene expression. In addition to enhancing accumulation of EBV gene mRNAs, SM has important effects on cellular mRNAs, altering the host cell gene expression profile to facilitate viral replication. This article describes the current state of knowledge regarding the role of EBV SM in cellular and viral gene regulation and summarizes some of the similarities and differences with the ORF57 homolog from Kaposi's sarcoma‐associated herpes virus (KSHV/HHV8). © 2005 Wiley‐Liss, Inc.
First‐principle studies of intermolecular and intramolecular catalysis of protonated cocaine
Authors: Zhan Chang‐Guo , Deng Shi‐Xian , Skiba Jaime G. , Hayes Beth A. , Tschampel Sarah M. , Shields George C. , Landry Donald W.
Published: May 2005

We have performed a series of first‐principles electronic structure calculations to examine the reaction pathways and the corresponding free energy barriers for the ester hydrolysis of protonated cocaine in its chair and boat conformations. The calculated free energy barriers for the benzoyl ester hydrolysis of protonated chair cocaine are close to the corresponding barriers calculated for the benzoyl ester hydrolysis of neutral cocaine. However, the free energy barrier calculated for the methyl ester hydrolysis of protonated cocaine in its chair conformation is significantly lower than for the methyl ester hydrolysis of neutral cocaine and for the dominant pathway of the benzoyl ester hydrolysis of protonated cocaine. The significant decrease of the free energy barrier, ∼4 kcal/mol, is attributed to the intramolecular acid catalysis of the methyl ester hydrolysis of protonated cocaine, because the transition state structure is stabilized by the strong hydrogen bond between the carbonyl oxygen of the methyl ester moiety and the protonated tropane N. The relative magnitudes of the free energy barriers calculated for different pathways of the ester hydrolysis of protonated chair cocaine are consistent with the experimental kinetic data for cocaine hydrolysis under physiologic conditions. Similar intramolecular acid catalysis also occurs for the benzoyl ester hydrolysis of (protonated) boat cocaine in the physiologic condition, although the contribution of the intramolecular hydrogen bonding to transition state stabilization is negligible. Nonetheless, the predictability of the intramolecular hydrogen bonding could be useful in generating antibody‐based catalysts that recruit cocaine to the boat conformation and an analog that elicited antibodies to approximate the protonated tropane N and the benzoyl O more closely than the natural boat conformer might increase the contribution from hydrogen bonding. Such a stable analog of the transition state for intramolecular catalysis of cocaine benzoyl‐ester hydrolysis was synthesized and used to successfully elicit a number of anticocaine catalytic antibodies. © 2005 Wiley Periodicals, Inc. J Comput Chem 26: 980–986, 2005
Thr 446 phosphorylation of PKR by HCV core protein deregulates G2/M phase in HCC cells
Authors: Alisi A. , Mele R. , Spaziani A. , Tavolaro S. , Palescandolo E. , Balsano C.
Published: May 2005

Hepatitis C virus (HCV) is the major causative viral agent of cirrhosis and hepatocarcinoma (HCC). HCV core protein affects cell homeostasis, playing an important role in viral pathogenesis of HCC. We investigate the effects of HCV core protein expression on cell growth in HCC cell lines. Cell cycle distribution analysis of HepG2 polyclonal core positive cells reveals a peculiar accumulation of cells in G2/M phase. Different pathways mediate G2/M arrest: such as p53 and double strand RNA protein kinase (PKR). Flow cytometry in p53‐null cells demonstrates that p53 plays only a marginal role in inducing HCV core‐dependent G2/M phase accumulation that seems to be significantly affected by the functional inactivation of PKR. HCC core positive cells are characterized by a significant PKR phosphorylation in Thr 446 residue, which leads deregulation of mitosis. Moreover, we observe that the overexpression of the viral protein induces an upregulation of PKR activity, which does not correlate with an increased eIF‐2 phosphorylation. This uncommon behavior of PKR suggests that its activation by HCV core protein could involve alternative PKR‐dependent pathways, implicated in core‐dependent G2/M accumulation. The described biological effects of HCV core protein on cell cycle could be an additional viral mechanism for both HCV resistance to interferon (IFN) and HCC HCV‐related pathogenesis. © 2005 Wiley‐Liss, Inc.
Inhibition of colon cancer cell proliferation by the dietary compound conjugated linoleic acid is mediated by the CDK inhibitor p21CIP1/WAF1
Authors: Lim Do Y. , Tyner Angela L. , Park Jae‐Bong , Lee Jae‐Yong , Choi Yung H. , Park Jung H.Y.
Published: May 2005

Our previous studies indicated that dietary conjugated linoleic acid (CLA) inhibits colon tumor cell proliferation in vitro and in vivo. To identify mechanisms by which CLA regulates growth arrest, the HT‐29 human colon carcinoma cell line was treated with various physiological concentrations of CLA and analyzed by flow cytometry. We detected a dose‐dependent increase in the percentage of cells arrested in G1 after CLA treatment that was accompanied by induction of the cyclin dependent kinase (CDK) inhibitor p21CIP1/WAF. CLA addition also led to increased p21 expression in HCT116 and SW480 cells, indicating that p21 induction is a general consequence of CLA treatment in colon cancer cells. Since both HT‐29 and SW480 cells have mutant p53, our data indicate that p53 is not essential for induction of p21. In addition to an increase in p21 levels, HT‐29 cell growth arrest was also accompanied by moderate decreases in Cyclin A, D1, E, and proliferating cell nuclear antigen (PCNA) levels. Following CLA treatment, p21 associated with and inhibited CDK4 and CDK2, and this correlated with reduced phosphorylation of retinoblastoma proteins. Increased association of p21 with PCNA was also detected. Dietary CLA inhibits cell cycle progression by inducing p21, which negatively regulates the growth promoting activities of CDK/cyclins and PCNA. These studies indicate that physiological concentrations of CLA inhibit growth of colon cancer cells with either wild‐type or mutant p53, and may have therapeutic benefits in vivo. © 2005 Wiley‐Liss, Inc.
Intrinsic excitability, synaptic potentials, and short‐term plasticity in human epileptic neocortex
Authors: Koch Uwe‐Robert , Mußhoff Ulrich , Pannek Heinz‐Wolfgang , Ebner Alois , Wolf Peter , Speckmann Erwin‐Josef , Köhling Rüdiger
Published: May 2005

Although studies of epileptic human hippocampus suggest changes of synaptic and intrinsic excitability, few changes, save the appearance of spontaneous field/synaptic potentials, are known in epileptic neocortical tissue. However, invasive EEG and histological studies suggest that neocortical tissue, even in mesial temporal lobe epilepsy, can play an important role as an irritative zone or extrahippocampal focus. We hypothesized that intrinsic neuronal and synaptic excitability, as well as short‐term plasticity, are altered in neocortical areas, particularly with elevated K+ levels as occur during seizures. We analyzed neuronal firing properties, synaptic responses, and paired‐pulse plasticity in human neocortical slices from tissue resected during epilepsy surgery, both under normal and under pathological conditions, i.e., after elevating K+ (4/8 mM), with rat neocortical slices as controls. Neuronal firing properties were not different. We did find, however, alterations of synaptic responsiveness in epileptic tissue, i.e., an elevated network excitability with K+ elevations, and reduction of paired‐pulse depression. © 2005 Wiley‐Liss, Inc.
Astrocytes protect neurons from ethanol‐induced oxidative stress and apoptotic death
Authors: Watts Lora Talley , Rathinam Mary Latha , Schenker Steven , Henderson George I.
Published: May 2005

Ethanol induces oxidative stress in cultured fetal rat cortical neurons and this is followed by apoptotic death, which can be prevented by normalization of cell content of reduced glutathione (GSH). Because astrocytes can play a central role in maintenance of neuron GSH homeostasis, the following experiments utilized cocultures of neonatal rat cortical astrocytes and fetal cortical neurons to determine if astrocytes could protect neurons from ethanol‐mediated apoptotic death via this mechanism. In cortical neurons cultured in the absence of astrocytes, ethanol (2.5 and 4 mg/ml; 6‐, 12‐, and 24‐hr exposures) decreased trypan blue exclusion and the MTT viability measures by up to 45% (P < 0.05), increased levels of reactive oxygen species (ROS) by up to 81% (P < 0.05), and decreased GSH within 1 hr of treatment by 49 and 51% for 2.5 and 4 mg/ml, respectively (P < 0.05). This was followed by onset of apoptotic cell death as determined by increased Annexin V binding and DNA fragmentation by 12 hr of ethanol exposure. Coculturing neurons with astrocytes prevented GSH depletion by 2.5 mg/ml ethanol, whereas GSH content was increased over controls in neurons exposed to 4 mg/ml ethanol (by up to 341%; P < 0.05). Ethanol generated increases in neuron ROS and apoptosis; decreases in viability were also prevented by coculture. Astrocytes were largely insensitive to ethanol, using the same measures. Only exposure to 4.0 mg/ml ethanol decreased GSH content in astrocytes, concomitant with a 204% increase in GSH efflux (P < 0.05). These studies illustrate that astrocytes can protect neurons from ethanol‐mediated apoptotic death and that this may be related to maintenance of neuron GSH. © 2005 Wiley‐Liss, Inc.
Effect of soybean addition on the rheological properties and breadmaking quality of wheat flour
Authors: Ribotta Pablo D , Arnulphi Sebastián A , León Alberto E , Añón María C
Published: May 2005

The effects of enzyme‐active full‐fat (EAFFSF), heat‐treated full‐fat (HTFFSF) and enzyme‐active defatted (EADSF) soy flours and commercial soy protein isolates (SPIs) on mixing properties and extensibility of dough, gluten formation, gas production and retention properties of dough and bread quality were studied. The soy products utilised in this study presented values of urease activity, nitrogen solubility index (NSI) and enthalpies of protein denaturation according to their previous heat processing. Soy products interfered in gluten formation, weakened dough strength and decreased dough gas retention capacity. Bread quality was negatively affected by soy product addition. The negative effects were exacerbated by an increase in soy/wheat ratio. Soybean protein state was identified as an important factor in determining dough and bread properties. Copyright © 2005 Society of Chemical Industry
Effect of saponins on the foam/flow properties of paste and physical characteristics of akara made from decorticated black‐eyed cowpeas
Authors: Park Jin‐Young , Plahar Minerva A , Hung Yen‐Con , McWatters Kay H , Eun Jong‐Bang
Published: May 2005

Akara, a deep‐fat‐fried cowpea paste product, is a popular snack food in West Africa. Foaming capacity of the cowpea paste is important for achieving a spongy light‐textured fried product. The effect of two saponins, Yucca and Quillaja, on the foaming capacity of cowpea paste was investigated. Results showed that an increase in the amount of saponin added (1, 2, 4%) resulted in a decrease in specific gravity and apparent viscosity of the cowpea paste. Addition of saponin also significantly reduced the firmness and increased the cohesiveness of the akara compared with the control. Akara made using the Quillaja saponin was less firm and more cohesive than akara made using the Yucca saponin. An increase in the amount of saponin added to the cowpea paste resulted in a darker (lower L*) and browner colour of akara. However, akara containing 1% Yucca saponin had similar colour attributes to the control but was less firm and more cohesive. Copyright © 2005 Society of Chemical Industry
Amplification and overexpression of Aurora kinase A (AURKA) in immortalized human ovarian epithelial (HOSE) cells
Authors: Chung C.M. , Man C. , Jin Y. , Jin C. , Guan X.Y. , Wang Q. , Wan T.S.K. , Cheung A.L.M. , Tsao S.W.
Published: May 2005

Immortalization is an early and essential step of human carcinogenesis. Amplification of chromosome 20q has been shown to be a common event in immortalized cells and cancers. We have previously reported that gain and amplification of chromosome 20q is a non‐random and common event in immortalized human ovarian surface epithelial (HOSE) cells. The chromosome 20q harbors genes including TGIF2 (20q11.2‐q12), AIB1 (20q12), PTPN1 (20q13.1), ZNF217 (20q13.2), and AURKA (20q13.2‐q13.3), which were previously reported to be amplified and overexpressed in ovarian cancers. Some of these genes may be involved in immortalization of HOSE cells and represent crucial premalignant changes in ovarian surface epithelium. Investigation of the involvement of these genes was examined in four pairs of pre‐crisis (preimmortalized) and post‐crisis (immortalized) HOSE cells. Overexpression of AURKA (Aurora kinase A), also known as BTAK and STK15, by both real time‐quantitative polymerase chain reaction (RT‐QPCR) and Western blotting was detected in all the four immortalized HOSE cells examined while overexpression of AIB1 and ZNF217 was observed in two of four immortalized HOSE cells examined. Overexpression of TGIF2 and PTPN1 was not significant in our immortalized HOSE cell systems. The degree of overexpression of AURKA was shown to be closely associated with the amplification of chromosome 20q in immortalized HOSE cells. Fluorescence in situ hybridization (FISH) with labeled P1 artificial clone (PAC) confirmed the amplification of the chromosomal region (20q13.2‐13.3) where AURKA resides. DNA amplification of AURKA was also confirmed using semi‐quantitative PCR. Our study showed that amplification and overexpression of AURKA is a common and significant event during immortalization of HOSE cells and may represent an important premalignant change in ovarian carcinogenesis. © 2005 Wiley‐Liss, Inc.
Facioscapulohumeral dystrophy presenting as infantile facial diplegia and late‐onset limb‐girdle myopathy in members of the same family
Authors: Felice Kevin J. , Jones Jennifer M. , Conway Stephen R.
Published: May 2005

We report a family with markedly variable myopathic weakness due to facioscapulohumeral muscular dystrophy (FSHD). The proband developed mild late‐onset proximal limb weakness. Her two daughters had severe infantile facial diplegia, initially diagnosed as Möbius syndrome, and mild childhood‐onset limb weakness and scapular winging. Results of facial muscle electromyography and muscle histopathology supported a myopathic disorder. This case study further highlights the broad clinical spectrum and intrafamily variability in FSHD, and the occasional absence of a positive correlation between fragment size and disease onset. Moreover, this study underscores the importance of considering FSHD in cases of infantile facial diplegia, especially in patients not demonstrating the full clinical features of Möbius syndrome. In difficult cases, facial muscle electromyography may help to differentiate myopathic from neuropathic weakness, and help guide further diagnostic studies. Muscle Nerve, 2005
Model reduction by constraints, discretization of flow problems and an induced pressure stabilization
Authors: Layton W.
Published: May 2005

For flow problems, turbulence models and numerical methods can be thought of as model reductions. Herein we consider reduction imposed as a constraint using Lagrange multipliers. Development of this last method recovers the subgrid eddy viscosity method and a new stabilization of the incompressibilty constraint. Analysis of the new incompressibility stabilization reveals that stabilization is only really needed for the finest pressure scales. Copyright © 2005 John Wiley & Sons, Ltd.