bluuebunny/crossref_metadata_2025 huggingface dataset:
Sleep-disordered breathing: a new risk factor of suspected fatty liver disease in overweight children and adolescents?
Authors: Verhulst S. L. , Schrauwen N. , Haentjens D. , Rooman R. P. , Van Gaal L. , De Backer W. A. , Desager K. N.
Published: September 2008

Sleep-disordered breathing (SDB) in childhood obesity is associated with hyperinsulinemia, dyslipidemia and inflammation and by these mechanisms, SDB could contribute to development of non-alcoholic fatty liver disease. We, therefore, investigated if SDB was an independent predictor of suspected fatty liver disease in a clinical sample of overweight and obese children and adolescents.

Retrospective case study of consecutive overweight or obese children and adolescents attending a paediatric obesity clinic. Suggestive fatty liver disease was defined as a serum alanine aminotransferase >40 U·L−1and/or a hyperechoic liver on abdominal ultrasound.

Subjects with suggestive fatty liver disease presented with higher waist circumference, more circulating peripheral leukocytes and a lower % of total sleep time with SaO2 ≥95% than their peers with a normal liver evaluation. Multiple logistic regression (stepwise forward) selected waist circumference (odds ratio = 1.05; 95% confidence interval = 1.00–1.10; p = 0.06) and SaO2nadir (odds ratio = 0.87; 95% confidence interval = 0.76–0.99; p = 0.03) as predictors of suggestive fatty liver disease.

This study suggests an association between the severity of SDB and suspected fatty liver disease in a clinical sample of overweight and obese children and adolescents. We strongly recommend more and carefully designed research on the influence of SDB on the development of fatty liver disease and on the effect of treating sleep apnoea on liver function parameters.
CPAP increases bronchial reactivity in OSAS patients
Authors: Korczyński P. , Górska K. , Bielicki P. , Byśkiniewicz K. , Zieliński J. , Chazan R.
Published: September 2008

Continuous positive airways pressure (CPAP) is a well known and safe method of treatment patients with obstructive sleep apnoea syndrome (OSAS). The effects of CPAP administration on the upper respiratory tract are known. However its effects on the lower respiratory tract still needs to be determined. Studies on bronchial hyperreactivity in patients treated by CPAP are contradictory.

The aim of the study was to assess the influence of a 3-week CPAP treatment in patients with OSAS and to evaluate associations between changes in bronchial reactivity and clinical features of OSAS and lung function tests (LFT).

Patients with newly diagnosed OSAS and lack of infection or chronic illness of the respiratory tract or other conditions which could influence bronchial hyperreactivity (BHR) were included.

Investigations were performed in 101 patients. There were 88 males and 13 females, mean age 51.5±11.2 years and BMI 32.6±5.4 kg·m−2. Qualified patients were randomly divided into 2 groups: 76 patients to CPAP treatment group, 25 control group. Both groups did not differ in anthropometrics features, severity of OSAS and LFT. Metacholine challenge test (MchCT) was performed at baseline and repeated after 3 weeks. Analysis of the individual results showed that in 11 patients the MchCT was positive (6 in the CPAP and 5 in the control groups). After 3 weeks in the group of CPAP treated patients an increase of BHR was noted. Log PC20M decreased from 1.38±0.3 to 1.26±0.5 (p<0.05). The number of patients with a positive result in the MchCT increased from 6 to 16 patients. There was no significant change in BHR in the control group.

It was found that CPAP treated patients with BHR were older, had less severe OSAS and lower FEV1 (p<0.05). In none of the patients positive result of BHR did no affect compliance to CPAP treatment.

Conclusions: CPAP therapy increases bronchial reactivity, but does not affect compliance to treatment.
The effect of baseline lung function on the determination of time to bronchiolitis obliterans syndrome
Authors: Burton Christopher M. , Iversen Martin , Carlsen Jørn , Andersen Claus , Mortensen Jann
Published: September 2008

Long term survival after lung transplantation depends on the development and severity of bronchiolitis obliterans syndrome (BOS).

The objective of this study was to identify the relationship between baseline FEV1and transplant procedure type, on the development and progression of BOS grade 0p to 3. All patients receiving a cadaveric lung transplant 1992-2004 were included in the study (n = 389). Exclusion criteria were patients surviving <3 months (n = 39) and missing spirometry measurements (n = 4).

There were significant differences between the transplant procedures, heart-lung (HLTx), double-lung (DLTx), and single-lung (SLTx), with respect to recipient age, BMI, and indication for transplantation. Baseline FEV1for HLTx (median 2.9L, quartiles 2.3–4.3L) and DLTx recipients (median 2.9L, quartiles 2.4–3.7L) were significantly larger than for patients undergoing SLTx procedures (median 1.6L, quartiles 1.3–1.9L), p<0.0001, respectively. Survival analyses demonstrated a significant association between baseline FEV1per litre and the development of BOS: grade 0p (HR: 0.59, CI: 0.51–0.68, p<0.0001); grade 1 (HR: 0.60, CI: 0.51–0.70. p<0.0001); grade 2 (HR: 0.62, CI 0.52–0.74, p<0.0001); and grade 3 (HR: 0.73, CI 0.60–0.89, p = 0.002). There was a significant log-log linear relationship between baseline FEV1and time to the development of BOS grades 0p to 3 in all patients developing the respective BOS grade (p<0.0001, respectively).

Baseline lung function is an important confounder and should be considered in future risk factor evaluations for the development and progression to BOS.
Monitoring the efficacy of anti-TB therapy by using the QuantiFERON-TB Gold In tube test
Authors: Markova R. , Drenska R. , Todorova Y. , Terzieva V. , Stefanova D.
Published: September 2008

Monitoring the efficacy of anti-TB therapy is crucial for the better control of the spread of MTB infection, but it is often difficult especially for patients without microbiological confirmation of the disease. IGRAs are novel indirect blood tests for M. tuberculosis infection and a promising marker of mycobacterial burden and disease activity.

To describe our experience with the use of the QuantiFERON-TB Gold In tube assay for monitoring of anti-TB therapy in patients with active disease 30 patients with active TB, all HIV-negative and BCG-vaccinated at birth, were studied before and after three and eight months of anti-TB therapy. The whole blood QFT G In tube assay was used to measure the MTB-specific IFN-gamma responses.

Before therapy 28/30 patients had positive results in QFT G In tube (QFT) and two – indeterminate. Three months after initiation of treatment all subjects tested were found positive. On month eight a significant decrease of MTB-specific responses was found in all patients but only 17/30 (56.7%) turned negative in QFT. Three of 13 patients with a positive response at the end of month eight, continued to have microbiological isolation and absence of clinical improvement (fig. 1, table 1).

On the basis of the results obtained, we concluded that the QFT Gold In tube assay could be used in routine clinical practice for monitoring anti-tuberculosis therapy. Further studies including larger number of patients and longer periods of observation are needed.
Lung defences: an overview: TABLE 1—
Authors: Nicod L. P.
Published: September 2008

Lung defences are dependant on a complex array of mechanisms in the upper airways, which must to be differentiated from those of the distal airways. However, the first lines of defence in the proximal and distal airways are predominantly based on mechanical barriers and several mechanisms related to innate immunity. If pathogens or antigens reach the interstitium, dendritic cells will take up these intruders, reporting antigenic information to the pulmonary lymph nodes, where an adaptive immunity will be generated. Dendritic cells, by doing so, bridge innate immunity with adaptive immunity. Knowledge of these mechanisms is key when modulating immunity to increase defence mechanisms or decrease allergic phenomena.
Improving inhaler use in COPD and the role of patient preference: FIGURE 1.
Authors: Bateman E. D.
Published: September 2008

Chronic obstructive pulmonary disease (COPD) is a difficult disease to manage, but recent research focusing on its pathophysiology has provided direction for the development of new treatments and improved management strategies.

COPD differs substantially from asthma, both in its pathophysiology and its treatment. Unlike asthma, COPD cannot be fully controlled or “reversed”; it is progressive and responses to anti-inflammatory drugs, including corticosteroids, have been disappointing, suggesting the presence of a unique, persistent form of inflammation. The current main emphasis in the treatment of COPD is therefore to minimise airflow obstruction using regular bronchodilator therapy. Apparently small improvements in traditional measures of airflow obstruction, such as functional residual capacity, may produce significant improvements in other measures, resulting in clinical benefit.

Ensuring efficient delivery of bronchodilators is vital to treatment success but has received little attention in guidelines to date. Inhaler technique, adherence rates and levels of satisfaction with therapy are all far from ideal. Improvements in these areas require more detailed consideration of the interactions between the patient, the healthcare provider and the inhaler device, and an examination of how inhaler choices are currently made by both healthcare provider and patient.
Patient needs and medication styles in COPD: FIGURE 1.
Authors: Osman L. M. , Hyland M. E.
Published: September 2008

Patient self-care in illnesses such as chronic obstructive pulmonary disease (COPD) is influenced by “trade-offs” that patients make between their social and psychological needs, and the “best practice” clinical management of their illness. Patients weigh the benefits of taking medication against the costs, such as symptoms.

Personal styles of medication use and decision-making also affect how well COPD patients respond to treatment plans and their acceptance of particular treatments, including inhalers. The large amount of information now available may have made patients more autonomous than before, but the actual locus of control varies widely between patients. Patients seem to take a pragmatic approach when assessing inhaler devices, basing preference on how effective they perceive the device to be. Patients with COPD show loyalty to devices and, in the main, try to comply with instructions given by the healthcare provider.

The health path for chronic obstructive pulmonary disease, which describes the course the disease typically takes, is a progressive deterioration. Patients therefore adopt a conservative attitude to their health goals and what they expect to gain from treatment and management plans. An understanding of these influences will help healthcare providers support the patient better and give the appropriate advice at each phase of a patient's health path.
Inhaler treatment options in COPD
Authors: Newman S. P.
Published: September 2008

A variety of inhaler devices are available for delivering treatments to patients with chronic obstructive pulmonary disease, and new inhalers are currently being developed. Each type of device has advantages and disadvantages, and the methods of preparation and use vary between them. The differences in instructions for use can easily confuse patients and health providers alike, resulting in incorrect use of many inhalers. “Crucial” errors in inhaler technique, whereby no drug is deposited in the lungs, must be avoided. Any type of inhaler can be misused so that little or no drug is deposited in the lungs.

It is now increasingly widely recognised that a successful treatment outcome in chronic obstructive pulmonary disease depends as much on the inhaler device as it does on the drug. Inhaler choice in chronic obstructive pulmonary disease should take into account whether the patient is likely to use it correctly, as well as patient preference and the likelihood of adherence to treatment.
In vivomodels of human airway epithelium repair and regeneration
Authors: Coraux C. , Hajj R. , Lesimple P. , Puchelle E.
Published: September 2008

Despite an efficient defence system, the airway surface epithelium, in permanent contact with the external milieu, is frequently injured by inhaled pollutants, microorganisms and viruses. The response of the airway surface epithelium to an acute injury includes a succession of cellular events varying from the loss of the surface epithelium integrity to partial shedding of the epithelium or even to complete denudation of the basement membrane. The epithelium has then to repair and regenerate to restore its functions. Thein vivostudy of epithelial regeneration in animal models has shown that airway epithelial cells are able to dedifferentiate, spread, migrate over the denuded basement membrane and progressively redifferentiate to reconstitute a functional respiratory epithelium after several weeks.

Humanised tracheal xenograft models have been developed in immunodeficient nude and severe combined immunodeficient (SCID) mice in order to mimic the natural regeneration process of the human airway epithelium and to analyse the cellular and molecular events involved during the different steps of airway epithelial reconstitution. These models represent very powerful tools for analysing the modulation of the biological functions of the epithelium during its regeneration. They are also very useful for identifying stem/progenitor cells of the human airway epithelium.

A better knowledge of the mechanisms involved in airway epithelium regeneration, as well as the characterisation of the epithelial stem and progenitor cells, may pave the way to regenerative therapeutics, allowing the reconstitution of a functional airway epithelium in numerous respiratory diseases, such as asthma, chronic obstructive pulmonary diseases, cystic fibrosis and bronchiolitis.
Costs of managing asthma as defined by a derived Asthma Control TestTMscore in seven European countries
Authors: Vervloet D. , Williams A. E. , Lloyd A. , Clark T. J. H.
Published: September 2008

The present authors explored the relationship between asthma control status, as measured by a derived Asthma Control TestTM(ACT) score, and the utilisation and cost of healthcare in Europe.

Data were derived from a European survey of asthma patients. Frequency of healthcare resource use was identified from the dataset and per-patient mean cost of asthma management estimated. Drug costs were not available. The ACT score was derived from questions in the survey identical or similar to the items comprising the ACT.

An ACT score was derived for 2,268 patients, of whom 48% (1,078) scored <20, suggesting their asthma was not well controlled, with 17% (381) scoring <15, suggesting poorly controlled asthma. The mean per-patient annual cost of asthma management for patients with a derived ACT of <15 was €1,604 (95% confidence interval: €1,219–2,084); for patients with a derived ACT score of 15–19, €512 (€404–660) and for patients with a derived ACT score of ≥20, €232 (€192–286). A higher derived ACT score was associated with significantly lower expenditure on asthma management.

Worse asthma control, as measured by the derived Asthma Control TestTMscore was associated with an increased requirement for unscheduled care and with higher cost.
YB-1 promotes microtubule assembly in vitro through interaction with tubulin and microtubules
Authors: Chernov Konstantin G , Mechulam Alain , Popova Nadezhda V , Pastre David , Nadezhdina Elena S , Skabkina Olga V , Shanina Nina A , Vasiliev Victor D , Tarrade Anne , Melki Judith , Joshi Vandana , Baconnais Sonia , Toma Flavio , Ovchinnikov Lev P , Curmi Patrick A
Published: September 2008

Background

YB-1 is a major regulator of gene expression in eukaryotic cells. In addition to its role in transcription, YB-1 plays a key role in translation and stabilization of mRNAs.

Results

We show here that YB-1 interacts with tubulin and microtubules and stimulates microtubule assembly in vitro. High resolution imaging via electron and atomic force microscopy revealed that microtubules assembled in the presence of YB-1 exhibited a normal single wall ultrastructure and indicated that YB-1 most probably coats the outer microtubule wall. Furthermore, we found that YB-1 also promotes the assembly of MAPs-tubulin and subtilisin-treated tubulin. Finally, we demonstrated that tubulin interferes with RNA:YB-1 complexes.

Conclusion

These results suggest that YB-1 may regulate microtubule assembly in vivo and that its interaction with tubulin may contribute to the control of mRNA translation.
Large-scale identification of polymorphic microsatellites using an in silico approach
Authors: Tang Jifeng , Baldwin Samantha J , Jacobs Jeanne ME , van der Linden C Gerard , Voorrips Roeland E , Leunissen Jack AM , van Eck Herman , Vosman Ben
Published: September 2008

Background

Simple Sequence Repeat (SSR) or microsatellite markers are valuable for genetic research. Experimental methods to develop SSR markers are laborious, time consuming and expensive.In silicoapproaches have become a practicable and relatively inexpensive alternative during the last decade, although testing putative SSR markers still is time consuming and expensive. In many species only a relatively small percentage of SSR markers turn out to be polymorphic. This is particularly true for markers derived from expressed sequence tags (ESTs). In EST databases a large redundancy of sequences is present, which may contain information on length-polymorphisms in the SSR they contain, and whether they have been derived from heterozygotes or from different genotypes. Up to now, although a number of programs have been developed to identify SSRs in EST sequences, no software can detect putatively polymorphic SSRs.

Results

We have developed PolySSR, a new pipeline to identify polymorphic SSRs rather than just SSRs. Sequence information is obtained from public EST databases derived from heterozygous individuals and/or at least two different genotypes. The pipeline includes PCR-primer design for the putatively polymorphic SSR markers, taking into account Single Nucleotide Polymorphisms (SNPs) in the flanking regions, thereby improving the success rate of the potential markers. A large number of polymorphic SSRs were identified using publicly available EST sequences of potato, tomato, rice,Arabidopsis,Brassicaand chicken.

The SSRs obtained were divided into long and short based on the number of times the motif was repeated. Surprisingly, the frequency of polymorphic SSRs was much higher in the short SSRs.

Conclusion

PolySSR is a very effective tool to identify polymorphic SSRs. Using PolySSR, several hundred putative markers were developed and stored in a searchable database. Validation experiments showed that almost all markers that were indicated as putatively polymorphic by polySSR were indeed polymorphic. This greatly improves the efficiency of marker development, especially in species where there are low levels of polymorphism, like tomato. When combined with the new sequencing technologies PolySSR will have a big impact on the development of polymorphic SSRs in any species.

PolySSR and the polymorphic SSR marker database are available fromhttp://www.bioinformatics.nl/tools/polyssr/.
Functional analyses reveal an important role for tyrosine residues in the staphylococcal multidrug efflux protein QacA
Authors: Wu Jingqin , Hassan Karl A , Skurray Ronald A , Brown Melissa H
Published: September 2008

Background

The staphylococcal QacA multidrug efflux protein confers resistance to an exceptional number of structurally unrelated antimicrobial compounds. Aromatic amino acid residues have been shown to be highly important for the transport function of several multidrug transporters and are intimately involved in multidrug binding. This study investigated the structural and functional importance of the seven tyrosine residues in QacA by examining the phenotypic effect of incorporating conservative (aromatic) and non-conservative (non-aromatic) substitutions for these residues.

Results

Determination of the resistance profiles and analysis of drug transport assays revealed that non-conservative substitutions for most tyrosine residues influenced the QacA drug recognition spectrum. However, an aromatic residue at three tyrosine positions, 63, 410 and 429, was of importance for QacA-mediated transport and resistance to the majority of substrates tested.

Conclusion

A tyrosine or phenylalanine residue at amino acid positions corresponding to 63 of QacA in related drug efflux proteins is found to be highly conserved. Therefore, an aromatic side chain at this position is likely to partake in a function common to these drug transporters, such as proton translocation or essential intramolecular contacts, whereas aromatic residues at the non-conserved 410 and 429 positions are expected to mediate a QacA-specific function, possibly forming or stabilising part of the QacA drug binding region.
Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR
Authors: Brackman Gilles , Defoirdt Tom , Miyamoto Carol , Bossier Peter , Van Calenbergh Serge , Nelis Hans , Coenye Tom
Published: September 2008

Background

To date, only few compounds targeting the AI-2 based quorum sensing (QS) system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in severalVibrio harveyimutants. We also studiedin vitrothe ability of these compounds to interfere with biofilm formation, stress response and virulence ofVibriospp. The compounds were also evaluated in anin vivoassay measuring the reduction ofVibrio harveyivirulence towardsArtemiashrimp.

Results

Our results indicate that cinnamaldehyde and several substituted derivatives interfere with AI-2 based QS without inhibiting bacterial growth. The active compounds neither interfered with the bioluminescence system as such, nor with the production of AI-2. Study of the effect in various mutants suggested that the target protein is LuxR. Mobility shift assays revealed a decreased DNA-binding ability of LuxR. The compounds were further shown to (i) inhibit biofilm formation in severalVibriospp., (ii) result in a reduced ability to survive starvation and antibiotic treatment, (iii) reduce pigment and protease production inVibrio anguillarumand (iv) protect gnotobioticArtemiashrimp against virulentVibrio harveyiBB120.

Conclusion

Cinnamaldehyde and cinnamaldehyde derivatives interfere with AI-2 based QS in variousVibriospp. by decreasing the DNA-binding ability of LuxR. The use of these compounds resulted in several marked phenotypic changes, including reduced virulence and increased susceptibility to stress. Since inhibitors of AI-2 based quorum sensing are rare, and considering the role of AI-2 in several processes these compounds may be useful leads towards antipathogenic drugs.
Comparative analysis of emmtype pattern of Group A Streptococcus throat and skin isolates from India and their association with closely related SIC, a streptococcal virulence factor
Authors: Sagar Vivek , Kumar Rajesh , Ganguly Nirmal K , Chakraborti Anuradha
Published: September 2008

Background

Group A streptococcus (GAS) causes a wide variety of life threatening diseases in humans and the incidence of such infections is high in developing countries like India. Although distribution of emm types of GAS in India has been described, there is a lack of data describing either the comparative distribution of emm types in throat versus skin isolates, or the distribution of certain virulence factors amongst these isolates. Therefore in the present study we have monitored the emm type pattern of Group A streptococcus throat and skin isolates from India. Additionally, the association of these isolates with closely related sic (crs), a multifunctional compliment binding virulence factor, was also explored.

Results

Of the 94 (46 throat and 48 skin) isolates analyzed, 37 emm types were identified. The most frequently observed emm types were emm49 (8.5%) and emm112 (7.5%) followed by 6.5% each of emm1-2, emm75, emm77, and emm81. Out of 37 emm types, 27 have been previously reported and rest were isolated for the first time in the Indian Community. The predominant emm types of throat (emm49 and emm75) samples were different from those of skin (emm44, emm81 and emm112) samples. After screening all the 94 isolates, the crs gene was found in six emm1-2 (crs1-2) isolates, which was confirmed by DNA sequencing and expression analysis. Despite the polymorphic nature of crs, no intravariation was observed within crs1-2. However, insertions and deletions of highly variable sizes were noticed in comparison to CRS isolated from other emm types (emm1.0, emm57). CRS1-2 showed maximum homology with CRS57, but the genomic location of crs1-2 was found to be the same as that of sic1.0. Further, among crs positive isolates, speA was only present in skin samples thus suggesting possible role of speA in tissue tropism.

Conclusion

Despite the diversity in emm type pattern of throat and skin isolates, no significant association between emm type and source of isolation was observed. The finding that the crs gene is highly conserved even in two different variants of emm1-2 GAS (speA +ve and -ve) suggests a single allele of crs may be prevalent in the highly diverse throat and skin isolates of GAS in India.
Effects of the N-methyl-D-Aspartate receptor antagonist dextromethorphan on vibrotactile adaptation
Authors: Folger Stephen E , Tannan Vinay , Zhang Zheng , Holden Jameson K , Tommerdahl Mark
Published: September 2008

Background

Previous reports have demonstrated that short durations of vibrotactile stimuli (less than or equal to 2 sec) effectively and consistently modify both the perceptual response in humans as well as the neurophysiological response in somatosensory cortex. The change in cortical response with adaptation has been well established by a number of studies, and other reports have extended those findings in determining that both GABA- and NMDAR-mediated neurotransmission play a significant role in the dynamic response of somatosensory cortical neurons. In this study, we evaluated the impact that dextromethorphan (DXM), an NMDAR antagonist, had on two distinct vibrotactile adaptation tasks.

Results

All subjects, both those that ingested 60 mg DXM and those that ingested placebo, were evaluated for their amplitude discriminative capacity between two simultaneously delivered vibrotactile stimuli both with and without 3 conditions of pre-exposure to adapting stimulation. The results demonstrated that the perceptual metrics of subjects who ingested 60 mg DXM were significantly altered from that of controls when the amplitude discrimination task followed one of the conditions of adapting stimulation. Without the condition of pre-exposure to an adapting stimulus (or stimuli), there was little difference between the observations obtained from the subjects that ingested DXM and controls. Peak impact on subject response occurred at 60 min post-ingestion, whereas the scores of controls who ingested placebo were not impacted.

Conclusion

The results – that DXM blocks vibrotactile adaptation – is consistent with the suggestion that NMDAR-mediated neurotransmission plays a significant role in the perceptual adaptive response. This finding is also consistent with neurophysiological findings that report observations of the effects of NMDAR block on the SI cortical response to repetitive vibrotactile stimulation.
Design and methods for a randomized clinical trial treating comorbid obesity and major depressive disorder
Authors: Schneider Kristin L , Bodenlos Jamie S , Ma Yunsheng , Olendzki Barbara , Oleski Jessica , Merriam Philip , Crawford Sybil , Ockene Ira S , Pagoto Sherry L
Published: September 2008

Background

Obesity is often comorbid with depression and individuals with this comorbidity fare worse in behavioral weight loss treatment. Treating depression directly prior to behavioral weight loss treatment might bolster weight loss outcomes in this population, but this has not yet been tested in a randomized clinical trial.

Methods and design

This randomized clinical trial will examine whether behavior therapy for depression administered prior to standard weight loss treatment produces greater weight loss than standard weight loss treatment alone. Obese women with major depressive disorder (N = 174) will be recruited from primary care clinics and the community and randomly assigned to one of the two treatment conditions. Treatment will last 2 years, and will include a 6-month intensive treatment phase followed by an 18-month maintenance phase. Follow-up assessment will occur at 6-months and 1- and 2 years following randomization. The primary outcome is weight loss. The study was designed to provide 90% power for detecting a weight change difference between conditions of 3.1 kg (standard deviation of 5.5 kg) at 1-year assuming a 25% rate of loss to follow-up. Secondary outcomes include depression, physical activity, dietary intake, psychosocial variables and cardiovascular risk factors. Potential mediators (e.g., adherence, depression, physical activity and caloric intake) of the intervention effect on weight change will also be examined.

Discussion

Treating depression before administering intensive health behavior interventions could potentially boost the impact on both mental and physical health outcomes.

Trial registration

NCT00572520
Factors associated with adolescent cigarette smoking in Greece: Results from a cross sectional study (GYTS Study)
Authors: Rachiotis George , Muula Adamson S , Rudatsikira Emmanuel , Siziya Seter , Kyrlesi Athina , Gourgoulianis Konstantinos , Hadjichristodoulou Christos
Published: September 2008

Background

Data about the predictors of smoking among adolescents in Greece are sparse. We tried to identify factors associated with current cigarette smoking among in-school adolescents in Greece in the context of GYTS study.

Methods

A secondary analysis of data from a questionnaire study using the Global Youth Tobacco Survey methodology was conducted to identify factors associated with smoking among adolescents in Greece. Data were collected in 2004–2005. The outcome variable was cigarette smoking within the past 30 days preceding the survey while independent variables included age, gender, parental educational status, parental smoking, perception of harmfulness of smoking, and the amount of pocket money at the adolescent's disposal.

Results

6141 adolescents (51.5% males and 48.5% females) participated in the study. In multivariate analysis, cigarette smoking was associated with male gender (OR: 1.62; 95% CI: 1, 08–3.08), parental smoking (OR: 2.59; 95% CI: 1.45–5.89), and having pocket money ≥ 16 Euros (OR: 2.64; 95% CI: 1.19–5.98).

Conclusion

Male gender, parental smoking, and having pocket-money ≥ 16 Euros were independently associated with current smoking among Greek students. These findings could be taken into account in order to formulate a comprehensive anti-smoking strategy in Greece.
Molecular analysis of chondrocytes cultured in agarose in response to dynamic compression
Authors: Bougault Carole , Paumier Anne , Aubert-Foucher Elisabeth , Mallein-Gerin Frédéric
Published: September 2008

Background

Articular cartilage is exposed to high mechanical loads under normal physiological conditions and articular chondrocytes regulate the composition of cartilaginous matrix, in response to mechanical signals. However, the intracellular pathways involved in mechanotransduction are still being defined. Using the well-characterized chondrocyte/agarose model system and dynamic compression, we report protocols for preparing and characterizing constructs of murine chondrocytes and agarose, and analyzing the effect of compression on steady-state level of mRNA by RT-PCR, gene transcription by gene reporter assay, and phosphorylation state of signalling molecules by Western-blotting. The mouse model is of particular interest because of the availability of a large choice of bio-molecular tools suitable to study it, as well as genetically modified mice.

Results

Chondrocytes cultured in agarose for one week were surrounded by a newly synthesized pericellular matrix, as revealed by immunohistochemistry prior to compression experiments. This observation indicates that this model system is suitable to study the role of matrix molecules and trans-membrane receptors in cellular responsiveness to mechanical stress. The chondrocyte/agarose constructs were then submitted to dynamic compression with FX-4000C™ Flexercell® Compression Plus™ System (Flexcell). After clearing proteins off agarose, Western-blotting analysis showed transient activation of Mitogen-activated protein kinases (MAPK) in response to dynamic compression. After assessment by capillary electrophoresis of the quality of RNA extracted from agarose, steady-state levels of mRNA expression was measured by real time PCR. We observed an up-regulation of cFos and cJun mRNA levels as a response to compression, in accordance with the mechanosensitive character observed for these two genes in other studies using cartilage explants submitted to compression. To explore further the biological response of mouse chondrocytes to the dynamic compression at the transcriptional level, we also developed an approach for monitoring changes in gene transcription in agarose culture by using reporter promoter constructs. A decrease in promoter activity of the gene coding for type II procollagen, the most abundant protein in cartilage, was observed in response to dynamic loading.

Conclusion

The protocols developed here offer the possibility to perform an integrated analysis of the molecular mechanisms of mechanotransduction in chondrocytes, at the gene and protein level.
In vivo effects of Pain Relieving Plaster on closed soft tissue injury in rabbit ears
Authors: Wang Yong-Zhi , Guo Chun-Yu , Zhong Hong-Gang , Zhang Wan-Nian , Wang De-Long , Wang Xuan , Dong Fu-Hui
Published: September 2008

Background

Soft tissue injury imposes major public health burdens worldwide. The positive effect of China's Tibetan medicine and theLamiophlomis rotata-based herbal Pain Relieving Plaster (PRP) on healing closed soft tissue injury (CSTI) has been reported. The herbs contained in Plaster are also referred as 'blood-activating and stasis-dispelling' in herbal medicine. The formula of the plaster contains four China's Tibetan medical herbs, includingLamiophlomis rotata,Oxytropis falcate Bunge,Curcuma longa Linn, andMyricaria bracteata. Two of these herbs (Lamiophlomis rotate;Curcuma longa Linn) are commonly used in different formulae of Chinese medicine. The objective of this study is to use an interdisciplinary approach to test the hypothesis that the formula and its components influence the process of CSTI.

Methods

In vivo models have been established in 30 rabbit ear pinnae and studied for: (1) blood flow velocity (BFV) which was affected by pressure of 21.2 kg/cm2 for 30 second over the local rabbit ear tissue; (2) edema formation of the closed soft tissue injury; (3) in vivo local temperature change.

Results

The results of in vivo studies indicated that CSTI significantly increased the velocity of blood flow and increased edema formation within the control group. The PRP extracts for 5 hours significantly slowed down the BFV of CSTI in rabbit ears, markedly decreased the elevated edema level from the 3rd to the 5th day.

Conclusion

The ingredients contained in the formula have positive effects in healing CSTI and further study is worth exploring.
Marine metagenomics: strategies for the discovery of novel enzymes with biotechnological applications from marine environments
Authors: Kennedy Jonathan , Marchesi Julian R , Dobson Alan DW
Published: September 2008

Metagenomic based strategies have previously been successfully employed as powerful tools to isolate and identify enzymes with novel biocatalytic activities from the unculturable component of microbial communities from various terrestrial environmental niches. Both sequence based and function based screening approaches have been employed to identify genes encoding novel biocatalytic activities and metabolic pathways from metagenomic libraries. While much of the focus to date has centred on terrestrial based microbial ecosystems, it is clear that the marine environment has enormous microbial biodiversity that remains largely unstudied. Marine microbes are both extremely abundant and diverse; the environments they occupy likewise consist of very diverse niches. As culture-dependent methods have thus far resulted in the isolation of only a tiny percentage of the marine microbiota the application of metagenomic strategies holds great potential to study and exploit the enormous microbial biodiversity which is present within these marine environments.
Use of the viral 2A peptide for bicistronic expression in transgenic mice
Authors: Trichas Georgios , Begbie Jo , Srinivas Shankar
Published: September 2008

Background

Transgenic animals are widely used in biomedical research and biotechnology. Multicistronic constructs, in which several proteins are encoded by a single messenger RNA, are commonly used in genetically engineered animals. This is currently done by using an internal ribosomal entry site to separate the different coding regions. 2A peptides result in the co-translational 'cleavage' of proteins and are an attractive alternative to the internal ribosomal entry site. They are more reliable than the internal ribosomal entry site and lead to expression of multiple cistrons at equimolar levels. They work in a wide variety of eukaryotic cells, but to date have not been demonstrated to function in transgenic mice in an inheritable manner.

Results

To test 2A function in transgenic mice and uncover any possible toxicity of widespread expression of the 2A peptide, we made a bicistronic reporter construct containing the coding sequence for a membrane localised red fluorescent protein (Myr-TdTomato) and a nuclear localised green fluorescent protein (H2B-GFP), separated by a 2A sequence. When this reporter is transfected into HeLa cells, the two fluorescent proteins correctly localise to mutually exclusive cellular compartments, demonstrating that the bicistronic construct is a reliable readout of 2A function. The two fluorescent proteins also correctly localise when the reporter is electroporated into chick neural tube cells. We made two independent transgenic mouse lines that express the bicistronic reporter ubiquitously. For both lines, transgenic mice are born in Mendelian frequencies and are found to be healthy and fertile. Myr-TdTomato and H2B-GFP segregate to mutually exclusive cellular compartments in all tissues examined from a broad range of developmental stages, ranging from embryo to adult. One transgenic line shows X-linked inheritance of the transgene and mosaic expression in females but uniform expression in males, indicating that the transgene has integrated into the X chromosome in this line.

Conclusion

The 2A peptide efficiently mediates co-translational cleavage in transgenic mice in which it has been inherited through the germ-line. Mice expressing it ubiquitously throughout development and into adulthood appear normal. It is therefore a viable tool for use in genetically engineered mice and represents a superior alternative to the widely used internal ribosomal entry site.
Sepsis causes neuroinflammation and concomitant decrease of cerebral metabolism
Authors: Semmler Alexander , Hermann Sven , Mormann Florian , Weberpals Marc , Paxian Stephan A , Okulla Thorsten , Schäfers Michael , Kummer Markus P , Klockgether Thomas , Heneka Michael T
Published: September 2008

Background

Septic encephalopathy is a severe brain dysfunction caused by systemic inflammation in the absence of direct brain infection. Changes in cerebral blood flow, release of inflammatory molecules and metabolic alterations contribute to neuronal dysfunction and cell death.

Methods

To investigate the relation of electrophysiological, metabolic and morphological changes caused by SE, we simultaneously assessed systemic circulation, regional cerebral blood flow and cortical electroencephalography in rats exposed to bacterial lipopolysaccharide. Additionally, cerebral glucose uptake, astro- and microglial activation as well as changes of inflammatory gene transcription were examined by small animal PET using [18F]FDG, immunohistochemistry, and real time PCR.

Results

While the systemic hemodynamic did not change significantly, regional cerebral blood flow was decreased in the cortex paralleled by a decrease of alpha activity of the electroencephalography. Cerebral glucose uptake was reduced in all analyzed neocortical areas, but preserved in the caudate nucleus, the hippocampus and the thalamus. Sepsis enhanced the transcription of several pro- and anti-inflammatory cytokines and chemokines including tumor necrosis factor alpha, interleukin-1 beta, transforming growth factor beta, and monocot chemoattractant protein 1 in the cerebrum. Regional analysis of different brain regions revealed an increase in ED1-positive microglia in the cortex, while total and neuronal cell counts decreased in the cortex and the hippocampus.

Conclusion

Together, the present study highlights the complexity of sepsis induced early impairment of neuronal metabolism and activity. Since our model uses techniques that determine parameters relevant to the clinical setting, it might be a useful tool to develop brain specific therapeutic strategies for human septic encephalopathy.
A high confidence, manually validated human blood plasma protein reference set
Authors: Schenk Susann , Schoenhals Gary J , de Souza Gustavo , Mann Matthias
Published: September 2008

Background

The immense diagnostic potential of human plasma has prompted great interest and effort in cataloging its contents, exemplified by the Human Proteome Organization (HUPO) Plasma Proteome Project (PPP) pilot project. Due to challenges in obtaining a reliable blood plasma protein list, HUPO later re-analysed their own original dataset with a more stringent statistical treatment that resulted in a much reduced list of high confidence (at least 95%) proteins compared with their original findings. In order to facilitate the discovery of novel biomarkers in the future and to realize the full diagnostic potential of blood plasma, we feel that there is still a need for an ultra-high confidence reference list (at least 99% confidence) of blood plasma proteins.

Methods

To address the complexity and dynamic protein concentration range of the plasma proteome, we employed a linear ion-trap-Fourier transform (LTQ-FT) and a linear ion trap-Orbitrap (LTQ-Orbitrap) for mass spectrometry (MS) analysis. Both instruments allow the measurement of peptide masses in the low ppm range. Furthermore, we employed a statistical score that allows database peptide identification searching using the products of two consecutive stages of tandem mass spectrometry (MS3). The combination of MS3 with very high mass accuracy in the parent peptide allows peptide identification with orders of magnitude more confidence than that typically achieved.

Results

Herein we established a high confidence set of 697 blood plasma proteins and achieved a high 'average sequence coverage' of more than 14 peptides per protein and a median of 6 peptides per protein. All proteins annotated as belonging to the immunoglobulin family as well as all hypothetical proteins whose peptides completely matched immunoglobulin sequences were excluded from this protein list. We also compared the results of using two high-end MS instruments as well as the use of various peptide and protein separation approaches. Furthermore, we characterized the plasma proteins using cellular localization information, as well as comparing our list of proteins to data from other sources, including the HUPO PPP dataset.

Conclusion

Superior instrumentation combined with rigorous validation criteria gave rise to a set of 697 plasma proteins in which we have very high confidence, demonstrated by an exceptionally low false peptide identification rate of 0.29%.
Identification of genes down-regulated during lung cancer progression: A cDNA array study
Authors: Campioni Mara , Ambrogi Vincenzo , Pompeo Eugenio , Citro Gennaro , Castelli Mauro , Spugnini Enrico P , Gatti Antonio , Cardelli Pierluigi , Lorenzon Laura , Baldi Alfonso , Mineo Tommaso C
Published: September 2008

Background

Lung cancer remains a major health challenge in the world. Survival for patients with stage I disease ranges between 40–70%. This suggests that a significant proportion of patients with stage I NSCLC may actually be under-staged.

Methods

In order to identify genes relevant for lung cancer development, we carried out cDNA array experiments employing 64 consecutive patients (58 men and 6 women) with a median age of 58 years and stage 1 or stage 2 non-small-cell lung cancer (NSCLC).

Results

Basic cDNA array data identified 14 genes as differentially regulated in the two groups. Quantitative RT-PCR analysis confirmed an effective different transcriptional regulation of 8 out of 14 genes analyzed. The products of these genes belong to different functional protein types, such as extra-cellular matrix proteins and proteases (Decorin and MMP11), genes involved in DNA repair (XRCC1), regulator of angiogenesis (VEGF), cell cycle regulators (Cyclin D1) and tumor-suppressor genes (Semaphorin 3B, WNT-5A and retinoblastoma-related Rb2/p130). Some previously described differences in expression patterns were confirmed by our array data. In addition, we identified and validated for the first time the reduced expression level of some genes during lung cancer progression.

Conclusion

Comparative hybridization by means of cDNA arrays assisted in identifying a series of novel progression-associated changes in gene expression, confirming, at the same time, a number of previously described results.