bluuebunny/crossref_metadata_2025 huggingface dataset:
Src kinases in systemic sclerosis: Central roles in fibroblast activation and in skin fibrosis
Authors: Skhirtladze Catherine , Distler Oliver , Dees Clara , Akhmetshina Alfiya , Busch Nicole , Venalis Paulius , Zwerina Jochen , Spriewald Bernd , Pileckyte Margarita , Schett Georg , Distler Jörg H. W.
Published: April 2008

Objective

Src kinases are nonreceptor tyrosine kinases, which have been implicated in cytoskeletal organization and cell mobility. This study was undertaken to evaluate the potential of Src kinases as novel targets of antifibrotic therapies.

Methods

Fibroblast cultures were obtained from 10 patients with systemic sclerosis (SSc) and 5 healthy subjects. Src signaling was inhibited using small‐molecule inhibitors and overexpression of a dominant‐negative mutant of Src and of the endogenous inhibitor Csk. The expression of extracellular matrix proteins was analyzed by real‐time polymerase chain reaction and by SirCol collagen assay. Toxic effects were excluded by MTT assay and staining for annexin V and propidium iodide. The mouse model of bleomycin‐induced dermal fibrosis was used to assess the role of Src kinases in dermal fibrosis in vivo.

Results

Stimulation with transforming growth factor β and platelet‐derived growth factor activated Src signaling in dermal fibroblasts from patients with SSc and healthy donors. Incubation with the Src kinase inhibitors or overexpressed mutant Src or Csk reduced the synthesis of messenger RNA for COL1A1, COL1A2, and fibronectin 1. A dose‐dependent reduction in collagen release was also observed at the protein level. No inhibitory effects on proliferation and no increase in the number of apoptotic or necrotic fibroblasts were observed. Consistent with the in vitro data, inhibition of Src kinases prevented experimental dermal fibrosis. Dermal thickness, the amount of collagen protein, and the number of myofibroblasts were reduced in a dose‐dependent manner.

Conclusion

These findings indicate that Src kinases play important roles in the activation of fibroblasts and in the development of experimental fibrosis. Thus, Src kinases might be interesting targets for novel antifibrotic therapies in SSc.
The pattern of response to anti–interleukin‐1 treatment distinguishes two subsets of patients with systemic‐onset juvenile idiopathic arthritis
Authors: Gattorno Marco , Piccini Alessandra , Lasigliè Denise , Tassi Sara , Brisca Giacomo , Carta Sonia , Delfino Laura , Ferlito Francesca , Pelagatti Maria Antonietta , Caroli Francesco , Buoncompagni Antonella , Viola Stefania , Loy Anna , Sironi Marina , Vecchi Annunciata , Ravelli Angelo , Martini Alberto , Rubartelli Anna
Published: April 2008

Objective

To assess the clinical response to interleukin‐1 (IL‐1) blockade and in vitro IL‐1β and IL‐18 secretion in patients with systemic‐onset juvenile idiopathic arthritis (JIA).

Methods

Twenty‐two patients with systemic‐onset JIA were treated with the IL‐1 receptor antagonist (IL‐1Ra) anakinra. Monocytes from 18 patients and 20 healthy donors were activated by different Toll‐like receptor ligands. Intracellular and secreted IL‐1β and IL‐18 were analyzed by Western blotting and enzyme‐linked immunosorbent assay.

Results

Ten patients with systemic‐onset JIA exhibited a dramatic response to anakinra and were classified as complete responders. Eleven patients had an incomplete response or no response, and 1 patient could not be classified in terms of response. Compared with patients who had an incomplete response or no response, complete responders had a lower number of active joints (P = 0.02) and an increased absolute neutrophil count (P = 0.02). In vitro IL‐1β and IL‐18 secretion in response to various stimuli was not increased and was independent of treatment efficacy. Likewise, secretion of IL‐1Ra by monocytes from patients with systemic‐onset JIA was not impaired. An overall low level of IL‐1β secretion upon exposure to exogenous ATP was observed, unrelated to treatment responsiveness or disease activity.

Conclusion

Two subsets of systemic‐onset JIA can be identified according to patient response to IL‐1 blockade. The 2 subsets appear to be characterized by some distinct clinical features. In vitro secretion of IL‐1β and IL‐18 by monocytes from patients with systemic‐onset JIA is not increased and is independent of both treatment outcome and disease activity.
Identification of the molecular response of articular cartilage to injury, by microarray screening: Wnt‐16 expression and signaling after injury and in osteoarthritis
Authors: Dell'Accio Francesco , De Bari Cosimo , Eltawil Noha M. , Vanhummelen Paul , Pitzalis Costantino
Published: April 2008

Objective

To characterize the molecular response of adult human articular cartilage to acute mechanical injury.

Methods

An established ex vivo model was used to compare gene expression of adult human articular cartilage explants 24 hours after mechanical injury with that of uninjured controls by microarray analysis of gene expression. Confirmation for selected genes was obtained by real‐time polymerase chain reaction and immunohistochemical analysis. Expression of selected genes was also investigated in preserved and osteoarthritic (OA) cartilage.

Results

Six hundred ninety genes were significantly regulated at least 2‐fold following mechanical injury. They included genes previously reported to be differentially expressed in OA versus normal cartilage or having allelic variants genetically linked to OA. Significant functional clusters included genes associated with wound healing, developmental processes, and skeletal development. The transforming growth factor β, fibroblast growth factor, and Wnt pathways were modulated. A systematic analysis of the Wnt signaling pathway revealed up‐regulation of Wnt‐16, down‐regulation of FRZB, up‐regulation of Wnt target genes, and nuclear localization of β‐catenin in injured cartilage. In addition, in OA, Wnt‐16 and β‐catenin were barely detectable in preserved cartilage areas, but were dramatically up‐regulated in areas of the same joint with moderate to severe OA damage.

Conclusion

Our findings indicate that mechanical injury to adult human articular cartilage results in the activation of a signaling response, with reactivation of morphogenetic pathways. Therapeutic targeting of such pathways may improve current protocols of joint surface defect repair and/or prevent the evolution of such lesions into posttraumatic OA.
Triple therapy in early active rheumatoid arthritis: A randomized, single‐blind, controlled trial comparing step‐up and parallel treatment strategies
Authors: Saunders S. A. , Capell H. A. , Stirling A. , Vallance R. , Kincaid W. , McMahon A. D. , Porter D. R.
Published: April 2008

Objective

The Tight Control of Rheumatoid Arthritis study previously demonstrated that an intensive step‐up disease‐modifying antirheumatic drug (DMARD) treatment strategy targeting persistent disease activity was superior to routine care in the management of early rheumatoid arthritis (RA). We undertook this study to test the hypothesis that early parallel triple therapy achieves better outcomes than step‐up therapy within an intensive disease management regimen.

Methods

Ninety‐six patients with early RA (mean disease duration 11.5 months) were randomized to receive step‐up therapy (sulfasalazine [SSZ] monotherapy, then after 3 months, methotrexate [MTX] was added, and when the maximum tolerated dosage of MTX was reached, hydroxychloroquine [HCQ] was added) or parallel triple therapy (SSZ/MTX/HCQ). All patients were assessed monthly for 12 months. If their disease activity score in 28 joints (DAS28) was ≥3.2, the dosage of DMARDs was increased according to protocol, and swollen joints were injected with triamcinolone acetonide (maximum dosage 80 mg per month). A metrologist who was blinded to the treatment allocation performed assessments every 3 months. The primary outcome measure was the mean decrease in the DAS28 score at 12 months.

Results

Both groups showed substantial improvements in disease activity and functional outcome. At 12 months, the mean decrease in the DAS28 score was −4.0 (step‐up therapy group) versus −3.3 (parallel therapy group) (P = 0.163). No significant differences in the percentages of patients with DAS28 remission (step‐up therapy group 45% versus parallel triple therapy group 33%), DAS28 good response (60% versus 41%, respectively), or American College of Rheumatology criteria for 20% improvement (ACR20) (77% versus 76%, respectively), ACR50 (60% versus 51%, respectively), or ACR70 (30% versus 20%, respectively) responses were seen. Radiologic progression was similar in both groups.

Conclusion

This study confirms that highly effective control of disease activity can be achieved using conventional DMARDs as part of an intensive disease management strategy. Within this setting, step‐up therapy is at least as effective as parallel triple therapy.
Assessment of unmet needs and the lack of generalizability in the design of randomized controlled trials for scleroderma treatment
Authors: Villela Renata , Yuen Sai Yan , Pope Janet E. , Baron Murray
Published: April 2008

Objective

To determine the generalizability of randomized controlled trials (RCTs) in the treatment of systemic sclerosis (SSc) using the Canadian Scleroderma Research Group (CSRG) database.

Methods

We identified articles related to SSc published from 1958 to 2006. Key points on trial design were recorded. The inclusion/exclusion criteria were used in conjunction with the CSRG database to determine the proportion of patients with SSc who would theoretically be eligible for these trials. Articles were classified into subcategories according to the target system. The CSRG database contains 438 patients with SSc from 14 Canadian centers. Results were in median (%) and mean (%) with 95% confidence intervals (95% CIs).

Results

In total, 210 articles were evaluated and 73 were selected for inclusion in this study. The mean percentage of eligible patients with SSc associated with other conditions was 35% (95% CI 17–53) for Raynaud's phenomenon, 24% (95% CI 1–47) for digital ulcers, 48% (95% CI 27–68) for gastrointestinal (GI) involvement, 32% (95% CI 20–43) for overall disease modification, 6% (95% CI 4–8) for pulmonary arterial hypertension, 2% (95% CI 0–4) for interstitial lung disease, and 38% (95% CI 12–64) for other categories.

Conclusion

Except for GI trials, <38% of the identified patients with SSc would have been suitable to enter the RCTs. Although some patients would be ineligible because they lack certain organ involvement, RCTs designed to include appropriate patients with SSc are needed; there are few proven treatments and trials typically do not include the majority of those who could potentially benefit from the intervention.
Use of the Human Activity Profile for estimating fitness in persons with arthritis
Authors: Bilek Laura D. , Venema Dawn M. , Willett Gilbert M. , Lyden Elizabeth R.
Published: April 2008

Objective

To evaluate the usefulness of the Human Activity Profile (HAP) in predicting estimated maximal oxygen uptake (VO2max) in persons with arthritis and to evaluate the ability of 2 classification systems to distinguish individuals with arthritis who have poor fitness from those with average and above fitness.

Methods

Forty‐four subjects with arthritis completed the HAP and a submaximal treadmill test. The adjusted activity score (AAS) was derived from responses on the HAP. VO2max was estimated from the submaximal treadmill test. The ability of the AAS and age to predict estimated VO2max was determined with multiple regression analysis. Subjects were also assigned to a fitness category based on their AAS and estimated VO2max, and agreement of these categories was assessed using the kappa statistic. Two classification systems were used, including one proposed by the original authors and one we proposed based on more recent normative data.

Results

Sixty‐six percent of the variance in estimated VO2max could be accounted for by the AAS and age. The kappa statistic for our proposed classification system was 0.35, indicating fair agreement, whereas the kappa statistic for the original classification system was incalculable. The sensitivity of the proposed classification system to identify persons with average and above fitness was 84%, with a specificity of 50%.

Conclusion

We suggest that the HAP is useful in estimating fitness level when standard exercise testing is not feasible.
Aortic involvement in recent‐onset giant cell (temporal) arteritis: A case–control prospective study using helical aortic computed tomodensitometric scan
Authors: Agard Christian , Barrier Jacques‐H. , Dupas Benoît , Ponge Thierry , Mahr Alfred , Fradet Gérard , Chevalet Pascal , Masseau Agathe , Batard Eric , Pottier Pierre , Planchon Bernard , Brisseau Jean‐Marie , Hamidou Mohamed‐A.
Published: April 2008

Objective

The prevalence of the involvement of large vessels in giant cell arteritis (GCA) is 3–13%. Aortitis is the most serious complication of GCA. Computed tomodensitometric (CT) scan allows analysis of both the aortic wall and endoluminal part of the aorta. Therefore, we conducted a study using CT scan to analyze aortic abnormalities in patients with recent‐onset GCA.

Methods

This prospective controlled study compared patients with biopsy‐proven GCA with a matched control group based on sex, age, and cardiovascular risk factors. During the 4‐week period following diagnosis of GCA, patients underwent an aortic CT scan. The aortic imaging results were blindly compared between both groups.

Results

From January 5, 1998 to January 11, 1999, 22 patients and 22 controls were screened by CT scan for aortic involvement. Thickening of the aortic wall was more frequent among patients than controls (45.4% versus 13.6%; P = 0.02). Aortic thickening (mean 3.3 mm) was located on the ascending part of the thoracic aorta in 22.7% of the patients, with no evidence of thickening in the controls (P = 0.05). Thickening of the abdominal aortic wall was noted in 27.3% of the patients and none of the controls (P = 0.02).

Conclusion

This study suggests that inflammatory aortic thickening, detected by CT scan, occurs frequently at the time of diagnosis of GCA, and that this condition predominantly occurs on the ascending part of the aorta.
Generalized Log‐Rank Tests for Partly Interval‐Censored Failure Time Data
Authors: Zhao Xingqiu , Zhao Qiang , Sun Jainguo , Kim Jong S.
Published: April 2008

In this paper, we consider incomplete survival data: partly interval‐censored failure time data where observed data include both exact and interval‐censored observations on the survival time of interest. We present a class of generalized log‐rank tests for this type of survival data and establish their asymptotic properties. The method is evaluated using simulation studies and illustrated by a set of real data from a diabetes study. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
A Relative Survival Model for Clustered Responses
Authors: Kuss Oliver , Blankenburg Thomas , Haerting Johannes
Published: April 2008

Relative Survival is the ratio of the overall survival of a group of patients to the expected survival for a demographically similar group. It is commonly used in disease registries to estimate the effect of a particular disease when the true cause of death is not reliably known. Regression models for relative survival have been described and we extend these models to allow for clustered responses by embedding them into the class of Generalized linear mixed models (GLMM). The method is motivated and demonstrated by a data set from the HALLUCA study, an epidemiological study which investigated provision of medical care to lung cancer patients in the region of Halle in the eastern part of Germany. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Understanding the Reactivity and Basicity of Zeolites: A Periodic DFT Study of the Disproportionation of N2O4 on Alkali‐Cation‐Exchanged Zeolite Y
Authors: Mignon Pierre , Pidko Evgeny A. , Van Santen Rutger A. , Geerlings Paul , Schoonheydt Robert A.
Published: April 2008

The disproportionation of N2O4 into NO3− and NO+ on Y zeolites has been studied through periodic DFT calculations to unravel 1) the role of metal cations and the framework oxygen atoms and 2) the relationship between the NO+ stretching frequency and the basicity of zeolites. We have considered three situations: adsorption on site II cations with and without a cation at site III and adsorption on a site III cation. We observed that cations at sites II and III cooperate to stabilize N2O4 and that the presence of a cation at site III is necessary to allow the disproportionation reaction. The strength of the stabilization is due to the number of stabilizing interactions increasing with the size of the cation and to the Lewis acidity of the alkali cations, which increases as the size of the cations decreases. In the product, NO3− interacts mainly with the cations and NO+ with the basic oxygen atoms of the tetrahedral aluminium through its nitrogen atom. As the cation size increases, the NO3−⋅⋅⋅cation interaction increases. As a result, the negative charge of the framework is less well screened by the larger cations and the interaction between NO+ and the basic oxygen atoms becomes stronger. NO+ appears to be a good probe of zeolite basicity, in agreement with experimental observations.
On the Mechanism of the Thermal Retrocycloaddition of Pyrrolidinofullerenes (Retro‐Prato Reaction)
Authors: Filippone Salvatore , Barroso Marta Izquierdo , Martín‐Domenech Ángel , Osuna Sílvia , Solà Miquel , Martín Nazario
Published: April 2008

In contrast to N‐methyl or N‐unsubstituted pyrrolidinofullerenes, which efficiently undergo the retrocycloaddition reaction to quantitatively afford pristine fullerene, N‐benzoyl derivatives do not give this reaction under the same experimental conditions. To unravel the mechanism of the retrocycloaddition process, trapping experiments of the in‐situ thermally generated azomethine ylides, with an efficient dipolarophile were conducted. These experiments afforded the respective cycloadducts as an endo/exo isomeric mixture. Theoretical calculations carried out at the DFT level and by using the two‐layered ONIOM (our own n‐layered integrated molecular orbital and molecular mechanics) approach underpin the experimental findings and predict that the presence of the dienophile is not a basic requirement for the azomethine ylide to be able to leave the fullerene surface under thermal conditions. Once the 1,3‐dipole is generated in the reaction medium, it is efficiently trapped by the dipolarophile (maleic anhydride or N‐phenylmaleimide). However, for N‐unsubstituted pyrrolidinofullerenes, the participation of the dipolarophile in assisting the 1,3‐dipole to leave the fullerene surface throughout the whole reaction pathway is also a plausible mechanism that cannot be ruled out.
Evidence‐based pathology and the pathologic evaluation of thymomas
Authors: Marchevsky Alberto M. , Gupta Ruta , McKenna Robert J. , Wick Mark , Moran Cesar , Zakowski Maureen F. , Suster Saul
Published: April 2008

BACKGROUND.

The clinical validity and applicability of the World Health Organization (WHO) histopathologic classification of thymomas (‘classification’) has been questioned. Evidence‐based pathology promotes the use of systematic reviews and analysis of data with meta‐analysis rather than subjective reviews of the literature.

METHODS.

The authors performed a review of the English literature from 1999 to the present to identify ‘best evidence’ regarding the use of the ‘classification.’ The data were analyzed with meta‐analysis software.

RESULTS.

To the authors' knowledge, only Level‐3 or ‐4 evidence published in retrospective cases series is currently available regarding the use of the ‘classification.’ Meta‐analysis demonstrated that only 3 WHO categories of thymomas are associated with significant survival differences: A/AB/B1, B2, and B3. It also indicated significant heterogeneity with regard to the results published in different studies. To the authors' knowledge there is no current evidence to determine whether thymoma types are significant prognostic features for patients previously stratified by stage.

CONCLUSIONS.

There is a lack of randomized clinical trials evaluating the prognosis of patients with thymomas and the effects of various treatment modalities. The WHO classification of thymomas needs revision and could most likely be simplified into fewer classes with significant prognostic value. Future studies are needed to evaluate the prognostic and/or predictive value for thymoma patients previously stratified by stage. The latter information is important to help select those patients who may benefit from neoadjuvant chemotherapy or postoperative radiotherapy and other modalities. Cancer 2008. © 2008 American Cancer Society.
Cervical cancer in patients infected with the human immunodeficiency virus
Authors: Leitao Mario M. , White Paul , Cracchiolo Bernadette
Published: April 2008

BACKGROUND.

The objective of this study was to compare the human immunodeficiency virus (HIV) viral load (VL) and CD4 counts in patients infected with HIV with and without cervical cancer. The authors hypothesized that HIV‐positive women with cervical cancer would have a greater risk of immune suppression.

METHODS.

A case–control study was conducted that included all HIV‐positive patients who were seen at the authors' institution from January 1, 1995 to April 20, 2006 with invasive cervical cancer (cases) and without invasive cervical cancer (controls). Patients were included only if they had a CD4 count recorded <6 months before or <3 months after their diagnosis of invasive cervical cancer (cases) or at their last gynecologic examination (controls). Controls were matched to cases on a 4:1 ratio according to current smoking history. Patients were considered immunocompetent if they had both a CD4 count >200 cells/μL and a VL <10,000 copies/mL.

RESULTS.

In total, 15 cases and 60 controls were identified. The median CD4 count for cases was 208 cells/μL (range, 18‐1102 cells/μL) compared with 445 cells/μL (range, 20‐1201 cells/μL) for controls (P = .03). The median VL was 16,918 copies/mL (range, 50‐214,915 copies/mL) for cases compared with 1430 copies/mL (range, 50‐571,000 copies/mL) for controls (P = .15). Only 1 of 14 cases (7%) was immunocompetent compared with 35 of 55 controls (64%; odds ratio, 0.04; 95% confidence interval, 0‐0.37; P < .001). This significance was maintained after adjusting for other factors (P = .002).

CONCLUSIONS.

Women with HIV who were diagnosed with invasive cervical cancer appeared to have a much greater degree of immunosuppression than women with HIV without invasive cervical cancer. Cancer 2008. © 2008 American Cancer Society.
Deamination of the Radical Cation of the Base Moiety of 2′‐Deoxycytidine: A Theoretical Study
Authors: Labet Vanessa , Grand André , Cadet Jean , Eriksson Leif A.
Published: April 2008

Five pathways leading to the deamination of cytosine (to uracil) after formation of its deprotonated radical cation are investigated in the gas phase, at the UB3LYP/6‐311G(d,p) level of theory, and in bulk aqueous solvent. The most favorable pathway involves hydrogen‐atom transfer from a water molecule to the N3 nitrogen of the deprotonated radical cation, followed by addition of the resulting hydroxyl radical to the C4 carbon of the cytosine derivative. Following protonation of the amino group (N4), the C4N4 bond is broken with elimination of the NH3⋅+ radical and formation of a protonated uracil. The rate‐determining step of this mechanism is hydrogen‐atom transfer from a water molecule to the cytosine derivative. The associated free energy barrier is 70.2 kJ mol−1.
Notch and suppressor of Hairless regulate levels but not patterns of Delta expression in Drosophila
Authors: Parks Annette L. , Shalaby Nevine A. , Muskavitch Marc A.T.
Published: April 2008

The Notch signal transduction pathway is highly conserved and governs many developmental decisions in metazoans. The ligand Delta, and its receptor Notch, are often expressed in complementary patterns during Drosophila postembryonic development. Notch signaling is thought to play a role in generation of these complementary patterns through feedback mechanisms that regulate Delta and Notch expression. We have examined Delta expression during postembryonic development, following global alteration of Notch‐dependent or Su(H)‐dependent transcriptional regulation. We find that Notch and Su(H) regulate Delta expression in a manner that varies by context. Surprisingly, we find that wild type Delta expression levels are influenced by Su(H)‐dependent mechanisms only in regions of high Delta/low Notch expression. In contrast, Delta expression levels in regions of low Delta/high Notch expression appear to be unaffected by Su(H)‐mediated regulation. We conclude that Notch pathway feedback regulation is unlikely to contribute to the generation of complementary patterns in the contexts examined. genesis 46:265–275, 2008. © 2008 Wiley‐Liss, Inc.
Use of fat replacers in the production of reduced‐calorie vanilla ice cream
Authors: Aykan Vildan , Sezgin Emel , Guzel‐Seydim Zeynep B.
Published: April 2008

The purpose of this research was to determine the effects of different fat replacers on the sensory and texture attributes of vanillin‐flavored ice cream. Inulin is a fairly new fat replacer which has been proven to be a health promoter. Two vanilla ice cream formulations were produced using different ratios of inulin and Simplesse® as fat replacers to provide better flavor and textural properties. Aspartame and acesulfam‐K were used as sweeteners in the low‐calorie ice creams. Sensory and instrumental analyses were used to evaluate the texture quality of light (4.55%) (control group), low‐fat (2%) and fat‐free vanilla ice creams. The experimental mixes were significantly more viscous than the control mix due to the water binding capacity of carbohydrate‐ and protein‐based substances. The texture scores of the ice creams (sensory evaluation) paralleled the viscosity values. Ice creams manufactured with fat replacers were free of cooked‐milk flavor. The energy value of ice cream was reduced compared to full‐fat ice cream (207 kcal/100 g): by 75% for non‐fat ice cream, 71% for low‐fat ice cream and 41% for light ice cream. An increase in total solids in the mix resulted in better texture, a lower melting rate and better shape retention.
Generation and Rearrangement of Some Spirocycloaliphatic Thiosulfines and Dithiiranes
Authors: Romański Jarosław , Reisenauer Hans Peter , Petzold Holm , Weigand Wolfgang , Schreiner Peter R. , Mlostoń Grzegorz
Published: April 2008

The flash‐vacuum pyrolysis of selected dispirodicycloaliphatic 1,2,4‐trithiolanes was studied by using matrix isolation techniques. The formation of thiosulfines and dithiiranes along with the corresponding thioketones was detected spectroscopically and confirmed by comparison with computed spectra. In the case of tetramethylcyclobutanone 1,2,4‐trithiolane, the thermal formation of the dithiolactone was observed. Pyrolysis of the (dichloro)tetramethylcyclobutane derivative resulted only in the formation of a mixture of the thiosulfine and dithiirane. In this case, conversion into the corresponding dithiolactone could be achieved photochemically. The dispirodiadamantane derivative gave a mixture of the respective thiosulfine and dithiirane but no dithiolactone could be detected in the mixture. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Protein extraction for proteome analysis from cacao leaves and meristems, organs infected by Moniliophthora perniciosa, the causal agent of the witches' broom disease
Authors: Pirovani Carlos Priminho , Carvalho Heliana Argôlo Santos , Machado Regina Cele Reboucas , Gomes Dayane Santos , Alvim Fátima Cerqueira , Pomella Alan William Vilela , Gramacho Karina Peres , Cascardo Júlio Cézar de Mattos , Pereira Gonçalo Amarante Guimarães , Micheli Fabienne
Published: April 2008

Preparation of high‐quality proteins from cacao vegetative organs is difficult due to very high endogenous levels of polysaccharides and polyphenols. In order to establish a routine procedure for the application of proteomic and biochemical analysis to cacao tissues, three new protocols were developed; one for apoplastic washing fluid (AWF) extraction, and two for protein extraction – under denaturing and nondenaturing conditions. The first described method allows a quick and easy collection of AWF – using infiltration–centrifugation procedure – that is representative of its composition in intact leaves according to the smaller symplastic contamination detected by the use of the hexose phosphate isomerase marker. Protein extraction under denaturing conditions for 2‐DE was remarkably improved by the combination of chemically and physically modified processes including phenol, SDS dense buffer and sonication steps. With this protocol, high‐quality proteins from cacao leaves and meristems were isolated, and for the first time well‐resolved 1‐DE and 2‐DE protein patterns of cacao vegetative organs are shown. It also appears that sonication associated with polysaccharide precipitation using tert‐butanol was a crucial step for the nondenaturing protein extraction and subsequent enzymatic activity detection. It is expected that the protocols described here could help to develop high‐level proteomic and biochemical studies in cacao also being applicable to other recalcitrant plant tissues.
Comparison of oil‐in‐water and water‐in‐oil microemulsion electrokinetic chromatography as methods for the analysis of eight phenolic acids and five diterpenoids
Authors: Cao Jun , Chen Jun , Yi Ling , Li Ping , Qi Lian‐Wen
Published: April 2008

Oil‐in‐water (O/W) and water‐in‐oil (W/O) MEEKC were compared for their abilities to separate and detect eight phenolic acids and five diterpenoids in Radix et Rhizoma Salviae Miltiorrhizae (RRSM). The effects of oil type and concentration, organic modifier, SDS, and buffer concentration on separation were examined in order to optimize the two methods. Oil contents and organic modifier were found to markedly influence the separation selectivity for both O/W and W/O systems. SDS concentration rarely affected separation resolution for O/W MEEKC, and separation of eight phenolic acids and five diterpenoids could be improved by changing the buffer concentration for W/O MEEKC. A highly efficient O/W MEEKC separation method, where the 13 compounds were separated with baseline resolution, was achieved by using a microemulsion solution of pH 8.0 containing 0.6% cyclohexane, 3.0% SDS, 6.0% 1‐butanol, and 3.0% ACN. The W/O MEEKC was unable to resolve all the components. In addition, the analytic time in O/W MEEKC was shorter than that in W/O MEEKC. Finally, the developed O/W MEEKC method was successfully applied to determine analytic compounds in RRSM samples.
Evidence of different stoichiometries for the limiting carbonate complexes across the lanthanide(III) series: A capillary electrophoresis‐mass spectrometry study
Authors: Philippini Violaine , Vercouter Thomas , Aupiais Jean , Topin Sylvain , Ambard Chrystel , Chaussé Annie , Vitorge Pierre
Published: April 2008

The electrophoretic mobilities (µep,Ln) of twelve lanthanides (not Ce, Pr and Yb) were measured by CE‐ICP‐MS in 0.15 and 0.5 mol L−1 Alk2CO3 aqueous solutions for Alk+ = Li+, Na+, K+ and Cs+. In 0.5 mol L−1 solutions, two different µep,Ln values were found for the light (La to Nd) and the heavy (Dy to Tm) lanthanides, which suggests two different stoichiometries for the carbonate limiting complexes. These results are consistent with a solubility study that attests the $\displaystyle {\tf=Ln}({\tf=CO}_3 )_3^{3 - }$ and $\displaystyle {\tf=Ln}({\tf=CO}_3 )_4^{5 - }$ stoichiometries for the heavy (small) and the light (big) lanthanides, respectively. The Alk+ counterions influence the $\displaystyle \mu _{{\tf=ep,Ln}}^{{\tf=Alk}_2 {\tf=CO}_3 }$ values, but not the overall shape of the $\displaystyle \mu _{{\tf=ep,Ln}}^{{\tf=Alk}_2 {\tf=CO}_3 }$ plots as a function of the lanthanide atomic numbers: the counterions do not modify the stoichiometries of the inner sphere complexes. The influence of the Alk+ counterions decreases in the Li+ > Na+ >> K+ > Cs+ series. The K3,Ln stepwise formation constants of the $\displaystyle {\tf=Ln}({\tf=CO}_3 )_3^{3 - }$ complexes slightly increase with the atomic numbers of the lanthanides while K4,Ln, the stepwise formation constants of $\displaystyle {\tf=Ln}({\tf=CO}_3 )_4^{5 - }$ complexes, slightly decrease from La to Tb, and is no longer measurable for heavier lanthanides.
Surgery and parental separation as potential risk factors for abnormal eating attitudes—longitudinal study
Authors: Bachar Eytan , Stein Daniel , Canetti Laura , Gur Eitan
Published: April 2008

Due to the susceptibility of eating disorders (ED) to stressful life events, we wanted to examine longitudinally whether two childhood adversities: (1) surgery and (2) parental separation, will affect abnormal eating attitudes in adolescents. Consecutively for 4 years, the eating attitude test (EAT‐26) and the eating disorder inventory‐2 (EDI‐2) questionnaires were administered to students from grades 7th through 10th and 8th through 11th. Multilevel analysis revealed that parental separation and oral or cosmetic dermatologic surgeries were significantly correlated with EAT‐26 and EDI‐2 scores throughout the 4 years of the study. Post‐hoc interpretation suggests a connection between (A) chirurgic intervention in the oral cavity and problematic eating attitudes, and (B) cosmetic dermatologic surgery and greater awareness to body appearance—a feature which might characterize adolescents who are prone to develop ED. Copyright © 2008 John Wiley & Sons, Ltd and Eating Disorders Association.
The natural volume method (NVM): Presentation and application to shallow water inviscid flows
Authors: Ata R. , Soulaïmani A. , Chinesta F.
Published: April 2008

In this paper a fully Lagrangian formulation is used to simulate 2D shallow water inviscid flows. The natural element method (NEM), which has been used successfully with several solid and fluid mechanics applications, is used to approximate the fluxes over Voronoi cells. This particle‐based method has shown huge potential in terms of handling problems involving large deformations. Its main advantage lies in the interpolant character of its shape function and consequently the ease it allows with respect to the imposition of Dirichlet boundary conditions.

In this paper, we use the NEM collocationally, and in a Lagrangian kinematic description, in order to simulate shallow water flows that are boundary moving problems. This formulation is ultimately shown to constitute a finite‐volume methodology requiring a flux computation on Voronoi cells rather than the standard elements, in a triangular or quadrilateral mesh. St Venant equations are used as the mathematical model. These equations have discontinuous solutions that physically represent the existence of shock waves, meaning that stabilization issues have thus been considered. An artificial viscosity deduced from an analogy with Riemann solvers is introduced to upwind the scheme and therefore stabilize the method. Some inviscid bidimensional flows were used as preliminary benchmark tests, which produced decent results, leading to well‐founded hopes for the future of this method in real applications. Copyright © 2008 John Wiley & Sons, Ltd.
Low‐affinity excitatory amino acid uptake in hippocampal astrocytes: A possible role of Na+/dicarboxylate cotransporters
Authors: Talgøy Holten Aleksander , Christian Danbolt Niels , Keiko Shimamoto , Vidar Gundersen
Published: April 2008

The excitatory amino acid transporters (EAATs) underlie the so‐called “high affinity” uptake of glutamate, which is well characterized. In contrast, the “low‐affinity” uptake of glutamate remains poorly defined, and it has been discussed whether it may represent a mere in vitro artifact. Here we have visualized “low‐affinity” excitatory amino acid uptake sites by incubating rat hippocampal slices with the glutamate analogue D‐aspartate in the presence of PMB‐TBOA, which blocks the EAATs. After fixation of the slices, D‐aspartate taken up into the tissue was localized with the use of light microscopic immunoperoxidase and electron microscopic immunogold methods, exploiting highly specific antibodies against D‐aspartate. PMB‐TBOA blocked uptake of both low and high exogenous D‐aspartate concentrations (0.01–1.0 mM) into nerve terminals, as well as the uptake of 0.01 mM D‐aspartate into astrocytes. Interestingly, there was a residual PMB‐TBOA insensitive uptake of D‐aspartate in astrocytes at higher exogenous D‐aspartate concentrations (0.05–1.0 mM), strongly suggesting that astrocytes have “low‐affinity” uptake sites for excitatory amino acid. The PMB‐TBOA insensitive D‐aspartate uptake in astrocytes was sodium dependent and inhibited by succinate and to certain extent by homocysteate, but not by cystine or DIDS. We suggest that excitatory amino acid is transported into astrocytes in a “low‐affinity” fashion by sodium/dicarboxylate transporters. © 2008 Wiley‐Liss, Inc.
Vasoactive intestinal peptide protects against β‐amyloid‐induced neurodegeneration by inhibiting microglia activation at multiple levels
Authors: Delgado Mario , Varela Nieves , Gonzalez‐Rey Elena
Published: April 2008

Alzheimer's disease (AD) is characterized by extracellular deposits of fibrillar β‐amyloid (Aβ) in the brain, increased microglial‐mediated inflammatory reactions in senile plaques, selective neuronal death, and cognitive deficits. The use of agents that limit microglial activation and inflammation in AD has recently emerged as an attractive therapeutic strategy for this disease. The vasoactive intestinal peptide (VIP), a widely distributed neuropeptide, has shown neuroprotective effects in acute brain damagein vivoand potent anti‐inflammatory actions in central nervous system. Here, we report that VIP inhibits Aβ‐induced neurodegeneration by indirectly inhibiting the production of a wide panel of inflammatory and neurotoxic agents by activated microglia cells. The inhibitory effect of VIP is mediated by blocking signaling through the p38 MAPK, p42/p44 MAPK, and NFkB cascades, the three major transduction pathways involved in the transcription of inflammatory mediators and the production of free radicals by Aβ‐activated microglia cells. Based on its neuroprotective action and its efficacy in inhibiting a broad range of inflammatory responses, VIP may provide a novel therapeutic approach to AD. © 2008 Wiley‐Liss, Inc.
Resting state sensorimotor functional connectivity in multiple sclerosis inversely correlates with transcallosal motor pathway transverse diffusivity
Authors: Lowe Mark J. , Beall Erik B. , Sakaie Ken E. , Koenig Katherine A. , Stone Lael , Marrie Ruth Ann , Phillips Micheal D.
Published: April 2008

Recent studies indicate that functional connectivity using low‐frequency BOLD fluctuations (LFBFs) is reduced between the bilateral primary sensorimotor regions in multiple sclerosis. In addition, it has been shown that pathway‐dependent measures of the transverse diffusivity of water in white matter correlate with related clinical measures of functional deficit in multiple sclerosis. Taken together, these methods suggest that MRI methods can be used to probe both functional connectivity and anatomic connectivity in subjects with known white matter impairment. We report the results of a study comparing anatomic connectivity of the transcallosal motor pathway, as measured with diffusion tensor imaging (DTI) and functional connectivity of the bilateral primary sensorimotor cortices (SMC), as measured with LFBFs in the resting state. High angular resolution diffusion imaging was combined with functional MRI to define the transcallosal white matter pathway connecting the bilateral primary SMC. Maps were generated from the probabilistic tracking employed and these maps were used to calculate the mean pathway diffusion measures fractional anisotropy 〈FA〉, mean diffusivity 〈MD〉, longitudinal diffusivity 〈λ1〉, and transverse diffusivity 〈λ2〉. These were compared with LFBF‐based functional connectivity measures (Fc) obtained at rest in a cohort of 11 multiple sclerosis patients and ∼10 age‐ and gender‐matched control subjects. The correlation between 〈FA〉 and Fc for MS patients was r = −0.63, P < 0.04. The correlation between all subjects 〈λ2〉 and Fc was r = 0.42, P < 0.05. The correlation between all subjects 〈λ2〉 and Fc was r = −0.50, P < 0.02. None of the control subject correlations were significant, nor were 〈FA〉, 〈λ1〉, or 〈MD〉 significantly correlated with Fc for MS patients. This constitutes the first in vivo observation of a correlation between measures of anatomic connectivity and functional connectivity using spontaneous LFBFs. Hum Brain Mapp, 2008. © 2008 Wiley‐Liss, Inc.
Fusion core structure of the severe acute respiratory syndrome coronavirus (SARS‐CoV): In search of potent SARS‐CoV entry inhibitors
Authors: Chu Ling‐Hon Matthew , Chan Siu‐Hong , Tsai Sau‐Na , Wang Yi , Cheng Christopher Hon‐Ki , Wong Kam‐Bo , Waye Mary Miu‐Yee , Ngai Sai‐Ming
Published: April 2008

Severe acute respiratory coronavirus (SARS‐CoV) spike (S) glycoprotein fusion core consists of a six‐helix bundle with the three C‐terminal heptad repeat (HR2) helices packed against a central coiled‐coil of the other three N‐terminal heptad repeat (HR1) helices. Each of the three peripheral HR2 helices shows prominent contacts with the hydrophobic surface of the central HR1 coiled‐coil. The concerted protein–protein interactions among the HR helices are responsible for the fusion event that leads to the release of the SARS‐CoV nucleocapsid into the target host‐cell. In this investigation, we applied recombinant protein and synthetic peptide‐based biophysical assays to characterize the biological activities of the HR helices. In a parallel experiment, we employed a HIV‐luc/SARS pseudotyped virus entry inhibition assay to screen for potent inhibitory activities on HR peptides derived from the SARS‐CoV S protein HR regions and a series of other small‐molecule drugs. Three HR peptides and five small‐molecule drugs were identified as potential inhibitors. ADS‐J1, which has been used to interfere with the fusogenesis of HIV‐1 onto CD4+ cells, demonstrated the highest HIV‐luc/SARS pseudotyped virus‐entry inhibition activity among the other small‐molecule drugs. Molecular modeling analysis suggested that ADS‐J1 may bind to the deep pocket of the hydrophobic groove on the surface of the central coiled‐coil of SARS‐CoV S HR protein and prevent the entrance of the SARS‐CoV into the host cells. J. Cell. Biochem. 104: 2335–2347, 2008. © 2008 Wiley‐Liss, Inc.
Photoionization cross sections with optimized orbital exponents within the complex basis function method
Authors: Morita Masato , Yabushita Satoshi
Published: April 2008

We show a new direction to expand the applicability of the complex basis function method for calculating photoionization cross sections through the imaginary part of the frequency‐dependent polarizability. Based on the variational stability of the frequency‐dependent polarizability, we made nonlinear optimizations of complex orbital exponents in basis functions representing continuum wave functions, and obtained fairly accurate results for H atom with only one or two complex basis functions particularly with dipole velocity gauge. Results were almost independent of whether Slater‐type or Gaussian‐type orbitals are used, implying the applicability to general many electron problems. The method was also applied to the 1S (1s)2 → 1P (1s)1(kp)1 cross section of He atom and the optimized complex orbital exponents were related to those of H atom through the scaling property. The nonlinear optimizations have converged smoothly and the cross sections were in excellent agreement with experiment throughout wide photon energies, which suggest the effectiveness of the approach for many‐electron systems. © 2008 Wiley Periodicals, Inc. J Comput Chem, 2008
Acute care hospital mortality of schizophrenic patients
Authors: Tokuda Yasuharu , Obara Haruo , Nakazato Nobuhiko , Stein Gerald H.
Published: April 2008

BACKGROUND:

Although schizophrenic patients have higher mortality than the general nonschizophrenic population, few studies have investigated acute care hospital mortality of schizophrenic patients.

METHODS:

To assess acute care hospital mortality of schizophrenic patients, we analyzed hospital mortality, the risk factors for hospital mortality, and the causes of death of schizophrenic patients admitted to an acute care hospital in Japan over an 18‐year period.

RESULTS:

We identified 55 deaths among 1108 schizophrenic patients, for a mortality rate of 5.0%. The overall standardized mortality ratio was 1.29 (95% CI, 0.98–1.68) compared with nonschizophrenic hospitalized patients. Significant risk factors for mortality were malignant neoplasm, cardiovascular disease, and admission through the emergency department. The leading causes of death in acute care hospitalizations were suicide (14.5%), malignant lymphoma or leukemia (10.9%), stroke (9.0%), and sepsis (7.3%).

CONCLUSIONS:

There was a nearly significant trend toward increased mortality of schizophrenic patients receiving acute hospital care compared with that of nonschizophrenic patients. Because suicide was the most frequent cause of death, a focus on suicide prevention is needed in this population. Journal of Hospital Medicine 2008;3:110–116. © 2008 Society of Hospital Medicine.
Innovative web‐based multimedia curriculum improves cardiac examination competency of residents
Authors: Criley Jasminka M. , Keiner Jennifer , Boker John R. , Criley Stuart R. , Warde Carole M.
Published: April 2008

BACKGROUND:

Proper diagnosis of cardiac disorders is a core competency of internists. Yet numerous studies have documented that the cardiac examination (CE) skills of physicians have declined compared with those of previous generations of physicians, attributed variously to inadequate exposure to cardiac patients and lack of skilled bedside teaching. With growing concerns about ensuring patient safety and quality of care, public and professional organizations are calling for a renewed emphasis on the teaching and evaluation of clinical skills in residency training.

OBJECTIVE:

The objective of the study was to determine whether Web training improves CE competency, whether residents retain what they learn, and whether a Web‐based curriculum plus clinical training is better than clinical training alone. Journal of Hospital Medicine 2008;3:124–133. © 2008 Society of Hospital Medicine.

DESIGN:

This was a controlled intervention study.

PARTICIPANTS:

The intervention group (34 internal and family medicine interns) participated in self‐directed use of a Web‐based tutorial and three 1‐hour teaching sessions taught by a hospitalist. Twenty‐five interns from the prior year served as controls.

MEASUREMENTS:

We assessed overall CE competency and 4 subcategories of CE competency: knowledge, audio skills, visual skills, and audio‐visual integration.

RESULTS:

The over mean score of the intervention group significantly improved, from 54 to 66 (P = .002). This improvement was retained (63.5, P = .05). When compared with end‐of‐year controls, the intervention group had significantly higher end‐of‐year CE scores (57 vs. 63.5, P = .05), knowledge (P = .04), and audio skills (P = .01). At the end of the academic year, all improvements were retained (P ≤ .04) except visual skills (P = .75).

CONCLUSIONS:

A Web‐based interactive educational program with minimal hospitalist involvement led to significant improvement in CE competence and was better than clinical training alone. © 2008 Society of Hospital Medicine.